The Global Isothermal Nucleic Acid Amplification Technology (INAAT) Market is expected to reach USD 4.3 billion by 2025, growing at a CAGR of 11.8%, according to a new report by Grand View Research, Inc. This can be attributed to significant rise in awareness regarding need for rapid and on-time testing for chronic and infectious diseases. Global increase in adoption of INAAT for detection of HIV, hepatitis A & B, and sexually transmitted diseases is considered to be a high impact rendering driver for growth.. Loop-Mediated Isothermal Amplification is one of the most preferred nucleic acid amplification methods because of its ability to provide point of care testing, thus leading to increased adoption. The recently-introduced Nicking Enzyme Amplification Reaction (NEAR) and Helicase-dependent amplification (HDA) technologies are expected to experience exponential growth during the forecast period owing to their patent protection.. In addition, owing to recent epidemics of infectious diseases, ...
The Global Isothermal Nucleic Acid Amplification Technology (INAAT) Market is expected to reach USD 4.3 billion by 2025, growing at a CAGR of 11.8%, according to a new report by Grand View Research, Inc. This can be attributed to significant rise in awareness regarding need for rapid and on-time testing for chronic and infectious diseases. Global increase in adoption of INAAT for detection of HIV, hepatitis A & B, and sexually transmitted diseases is considered to be a high impact rendering driver for growth.. Loop-Mediated Isothermal Amplification is one of the most preferred nucleic acid amplification methods because of its ability to provide point of care testing, thus leading to increased adoption. The recently-introduced Nicking Enzyme Amplification Reaction (NEAR) and Helicase-dependent amplification (HDA) technologies are expected to experience exponential growth during the forecast period owing to their patent protection.. In addition, owing to recent epidemics of infectious diseases, ...
The Global Isothermal Nucleic Acid Amplification Technology (INAAT) Market is expected to reach USD 4.3 billion by 2025, growing at a CAGR of 11.8%, according to a new report by Grand View Research, Inc. This can be attributed to significant rise in awareness regarding need for rapid and on-time testing for chronic and infectious diseases. Global increase in adoption of INAAT for detection of HIV, hepatitis A & B, and sexually transmitted diseases is considered to be a high impact rendering driver for growth.. Loop-Mediated Isothermal Amplification is one of the most preferred nucleic acid amplification methods because of its ability to provide point of care testing, thus leading to increased adoption. The recently-introduced Nicking Enzyme Amplification Reaction (NEAR) and Helicase-dependent amplification (HDA) technologies are expected to experience exponential growth during the forecast period owing to their patent protection.. In addition, owing to recent epidemics of infectious diseases, ...
The Global market for Isothermal Nucleic Acid Amplification Technology is poised to reach $2.1 billion by the end of 2020 growing at a CAGR of 10.5%. The P
Rapid and Sensitive Detection of Salmonella spp. by Using a Loop-Mediated Isothermal Amplification Assay in Duck Carcass Sample - loop-mediated isothermal amplification;Salmonella spp.;screening;duck;
A Loop-Mediated Isothermal Amplification Assay and Sample Preparation Procedure for Sensitive Detection of Xanthomonas fragariae in Strawberry. . Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.
Read "Detection of hepatitis C virus by an improved loop-mediated isothermal amplification assay, Archives of Virology" on DeepDyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips.
This study was designed to optimize and apply the use of loop-mediated isothermal amplification (LAMP) as an alternative to conventional polymerase chain reaction (PCR) for the detection of herpesvirus of turkeys (HVT) (FC 126 strain) in vaccinated and non-vaccinated poultry in Nigeria
BioMed Research International is a peer-reviewed, Open Access journal that publishes original research articles, review articles, and clinical studies covering a wide range of subjects in life sciences and medicine. The journal is divided into 55 subject-specific sections.
Chlamydiae are implicated in a variety of clinically and economically important diseases in livestock and companion animals. These bacteria are associated with abortion, conjunctivitis, encephalomyelitis, enteritis, pneumonia, and polyarthritis in ruminants. Infection with these bacteria is the most common cause of abortion in sheep and goats and also causes zoonotic infection in humans which, in pregnant women, can result in spontaneous abortion.
Abstract. Monitoring post-control transmission of schistosomes by examining humans becomes less effective as infection rates among humans decrease. Molecular monitoring of prepatent schistosome infection in snails by the polymerase chain reaction (PCR) has been used for studying human-to-snail transmission, and snail prepatent infection rates were found to correspond to infection prevalence and average intensity in human populations contacting the sites studied. We have now developed loop-mediated isothermal amplification (LAMP) assays for identifying Schistosoma mansoni and S. haematobium to facilitate large-scale evaluation of post-intervention transmission potential. LAMP primers were designed based on the Sm1-7 and DraI repeated sequences of the corresponding schistosomes, and amplification by LAMP of these 121-basepair highly abundant sequences provided a detection sensitivity of 0.1 fg of genomic DNA. When these LAMP assays were applied for examining infected laboratory snails, it was possible to
Because of the serological cross-reactivity among the flaviviruses, molecular detection methods, such as reverse-transcription polymerase chain reaction (RT-PCR), play an important role in the recent Zika outbreak. However, due to the limited sensitivity, the detection window of RT-PCR for Zika viremia is only about one week after symptom onset. By combining loop-mediated isothermal amplification (LAMP) and AC susceptometry, we demonstrate a rapid and homogeneous detection system for the Zika virus oligonucleotide. Streptavidin-magnetic nanoparticles (streptavidin-MNPs) are premixed with LAMP reagents including the analyte and biotinylated primers, and their hydrodynamic volumes are dramatically increased after a successful LAMP reaction. Analyzed by a portable AC susceptometer, the changes of the hydrodynamic volume are probed as Brownian relaxation frequency shifts, which can be used to quantify the Zika virus oligonucleotide. The proposed detection system can recognize 1 aM synthetic Zika ...
A loop-mediated isothermal amplification (LAMP) assay for open reading frame 1 (ORF1) of the glutamine synthetase gene of Neisseria gonorrhoeae was able to tolerate urea concentrations of ≤1.8 M, compared with a PCR assay that was functional at concentrations of ,100 mM. The LAMP assay was as sensitive as the PCR assay while being faster and simpler to perform.. ...
As a first step in developing a quick, accurate and simple method for the diagnosis of red ring disease, the loop-mediated isothermal amplification (LAMP)-based identification procedure was applied to the causative agent, Bursaphelenchus cocophilus. Two LAMP primer sets were designed using two loci of ribosomal RNA genes, i.e., D2-D3 expansion segments of the large subunit (D2-D3 LSU), and internal transcribed spacers (ITS). Within those two sets of primers, the D2-D3 LSU primer set successfully yielded amplicons from B. cocophilus nematode lysate prepared from 3-year-old DESS-fixed specimens. The specificity of the primers was examined using 18 species of confamilial Aphelenchoididae nematodes and primer sensitivity was tested using a diluted series of B. cocophilus lysate. The primer set did not amplify the DNA from other aphelenchoidids, and sensitivity was achieved by 1:100 diluted B. cocophilus DNA (roughly 1/1500 of total DNA from a single third-stage juvenile).
In this study, a loop-mediated isothermal amplification (LAMP) assay was established to detect Toxoplasma gondii DNA in mice infected with T. gondii PRU strain. This LAMP assay was based on the sequence of highly repetitive B1 gene. The detect...
Laboratory diagnosis has relied on the identification of malaria parasites and parasite antigens in peripheral blood using Giemsa-stained microscopy or rapid diagnostic tests (RDTs) which lack analytical and clinical sensitivity. The aim of this study was to evaluate the performance of loop-mediated isothermal amplification (LAMP) for the diagnosis of malaria among malaria suspected pregnant women in Northwest Ethiopia. The authors conclude that this study showed higher sensitivity of LAMP compared to microscopy and RDT for the detection of malaria in pregnancy. Increased sensitivity and ease of use with LAMP in point-of-care testing for malaria in pregnancy was noted. LAMP warrants further evaluation in intermittent screening and treatment programmes in pregnancy.. ...
Vet Parasitol. 2013 Feb 18;192(1-3):98-103. doi: 10.1016/j.vetpar.2012.10.010. Epub 2012 Oct 23. Research Support, Non-U.S. Govt
In aquaculture, vibriosis is known as a major bacterial disease in fish culture systems and can cause considerable loss in terms of production and processing (Toranzo et al. 2005). Many Vibrio species have been recognized as fish pathogens that can cause infection with various symptoms. For example, Vibrio scophthalmi infection results in hemorrhage on fish body surface and inner surface of the abdomen, severe enteritis, and ascites (Qiao et al. 2012); Vibrio ichthyoenteri infection cause opaque intestines and necrotizing fasciitis with high mortality rates (Ishimaru et al. 1996; Lee et al. 2012); Vibrio parahaemolyticus infection causes diseases not only in fish, shrimp, oysters, and mussels, etc. (Montilla et al. 1994; Quintoil et al. 2007), but also is important in public health and causes gastrointestinal disorders in humans who ingest contaminated fish and shellfish (Kubota et al. 2008; Iwamoto et al. 2010); and Vibrio vulnificus has been associated with vibriosis outbreaks in fish and ...
Battling infection is a major healthcare objective. Untreated infections can rapidly evolve toward the condition of sepsis in which the body begins to fail and resuscitation becomes critical and tenuous. Identification of infection followed by rapid antimicrobial treatment are primary goals of medical care, but precise identification of offending organisms by current methods is slow and broad spectrum empirical therapy is employed to cover most potential pathogens. Current methods for identification of bacterial pathogens in a clinical setting typically require days of time, or a four- to eight-hour growth phase followed by DNA extraction, purification and PCR-based amplification. We demonstrate rapid (70-120 minutes) genetic diagnostics methods utilizing loop-mediated isothermal amplification (LAMP) to test for 15 common infection pathogen targets, called the Infection Diagnosis Panel (In-Dx). The method utilizes filtration to rapidly concentrate bacteria in sample matrices with lower bacterial loads
In this study, we constructed a novel tool for the identification of six Plasmodium parasites, including two P. ovale subspecies, using MinION™ portable sequencer and the LAMP method. We demonstrated that the method can achieve a comprehensive diagnosis of malaria even in resource-limited endemic regions. In particular, the LAMP method employs a set of four specially designed primers that recognize six distinct sequences on the target DNA, and relies on an auto-cycling procedure under isothermal conditions. In addition, our method enabled rapid library preparation (within 30 min), with multiplex sequencing and streaming analysis of real-time sequencing data via the Rapid Barcoding Sequencing Kit. Therefore, our LAMP method combined with the MinION™ sequencer would be convenient to use in various clinical settings.. Recently, Loopamp™ MALARIA Pan/Pf Detection Kit (Eiken Chemical) has become commercially available for the diagnosis of malaria by the LAMP method. This LAMP kit has good ...
0002] Nucleic acid amplification technologies, such as a polymerase chain reaction (hereinafter, referred to as "PCR") that requires a temperature change in reaction liquid amplification and detection processes and a loop mediated isothermal amplification method (hereinafter, referred to as "LAMP method") that does not require a temperature change in reaction liquid amplification and detection processes, have been used for amplification and quantification of nucleic acids contained in samples originating in living organisms. For nucleic acid amplification, PCR requires periodically changing a sample temperature in typically about two to three temperature regions. For example, a typical PCR is performed as follows: the sample temperature is heated up to 94° C. to separate the double strands, followed by annealing at 60° C., and the sample temperature is kept at 60° C. to 72° C. for a few minutes. This PCR process is repeated by n times to amplify a target nuclear acid. On the other hand, in ...
A47 Recently, GD-100 (SYSMEX, Kobe, Japan) has been released. It rapidly can detect and quantify mRNA by one-step nucleic acid amplification (OSNA). Loop-mediated isothermal amplification (LAMP) is performed and turbidity derived from magnesium pyrophosphate formation with LAMP can be monitored in OSNA. LAMP originally was reported in 2000 as a novel nucleic acid amplification method that uses only one type of enzyme and yields 109 of its target in less than an hour. In this study, we compared the sensitivity of OSNA using GD-100 versus conventional real-time RT-PCR, using five colon cancer cell lines. We then applied OSNA to detect micro-metastases in 1) lymph nodes (LNs) from patients with advanced colorectal adenocarcinomas; and 2) LNs submitted for frozen diagnosis from patients with several malignancies. Our goal was to develop a more efficient system to detect LN metastases than histological examination (HE) performed intra-operatively.Methods: CK19 mRNA copy numbers of five colon ...
Multiple displacement amplification (MDA) is a widely used technique for amplification of DNA from samples containing limited amounts of DNA (e.g., uncultivable microbes or clinical samples) before whole genome sequencing. Despite its advantages of high yield and fidelity, it suffers from high amplification bias and non-specific amplification when amplifying sub-nanogram of template DNA. Here, we present a microfluidic digital droplet MDA (ddMDA) technique where partitioning of the template DNA into thousands of sub-nanoliter droplets, each containing a small number of DNA fragments, greatly reduces the competition among DNA fragments for primers and polymerase thereby greatly reducing amplification bias. Consequently, the ddMDA approach enabled a more uniform coverage of amplification over the entire length of the genome, with significantly lower bias and non-specific amplification than conventional MDA. For a sample containing 0.1 pg/μL of E. coli DNA (equivalent of ~3/1000 of an E. coli genome per
Currently available diagnostic approaches for tuberculosis are limited by low sensitivity and time factors. The present gold standard for diagnosis is isolation of positive culture from clinical specimens, which generally takes up to 8 weeks for results. Unfortunately, even with rapid culture techniques, the results take at least 1 week to return, which increases the difficulty of identifying infected patients and ensuring that they follow-up for treatment. Nucleic acid amplification techniques are limited by their sensitivity; they are presently widely recommended only for AFB-smear (+) patients. The EliSpot assay, which detects IFN-g producing T cells specific for mTB, cannot reliably distinguish between active and latent TB given that the test is performed on peripheral blood samples. Therefore, rapid, highly sensitive tests are desperately needed in order to identify patients infected with TB and limit the transmission of this potentially fatal infection. We have developed an innovative ...
3. 233 Hepatitis C virus (HCV), validation of nucleic acid amplification techniques for the detection streett HCV RNA in plasma pools Guidelines.
Molecular diagnostics refers to a technique used to detect and identify the presence of genetic material or proteins associated with a specific health condition or disease. Viral molecular diagnostic helps in diagnosis of infectious diseases caused by virus. Viral molecular diagnostics has varied applications in hospitals, academics institutions, laboratories and others. On the basis of application, viral molecular diagnostics can be segmented into infectious diseases, genetics, blood screening, microbiology and others. On the basis of infectious diseases, viral molecular diagnostics market can be segmented into hepatitis B virus (HBV), hepatitis C virus (HCV), human immunodeficiency virus (HIV) human papillomavirus (HPV) and others. On the basis of technology, viral molecular diagnostics market can be segmented into polymerase chain reaction (PCR), isothermal nucleic acid amplification technology (INAAT), microarrays, in situ hybridization and others.. North America, followed by Europe, has the ...
Sampling technique and detection rates of oropharyngeal and anorectal gonorrhoea using nucleic acid amplification tests in men who have sex with men ...
We tested our hypothesis that double strand breaks can improve the loading efficiency of the helicase near a target sequence by evaluating the impact of restriction endonuclease digestion during HDA amplification. As shown in Figure 1B, specific restriction enzyme can be selected to cleave a specific sequence near the target sequence, generate 5ss (single-stranded) ends, 3ss ends or blunt ends to help recruiting and loading the helicase. Therefore, one simple protein for mediation of helicase homing can replace the functions of multiple accessory proteins in vivo. Indeed, previous studies had already used restriction enzymes to produce substrates for helicase unwinding assay in vitro [12, 13]. So far more than 3000 restriction endonucleases with over two hundred different specificities have been isolated from bacteria [14]. This broad collection makes finding an enzyme which can cut the DNA duplex close to the target very straightforward. The "time-saver qualified enzymes" from New England ...
A unit dose test device for a nucleic acid amplification reaction has the form of a elongate disposable test strip. The test strip includes a dual-chamber reaction vessel pre-loaded with nucleic acid amplification reaction reagents, and a plurality of wells for processing a reaction occurring in the reaction vessel.
Genetically modified (GM) rice KMD1, TT51-1, and KF6 are three of the most well known transgenic Bt rice lines in China. A rapid and sensitive molecular assay for risk assessment of GM rice is needed. Polymerase chain reaction (PCR), currently the most common method for detecting genetically modified organisms, requires temperature cycling and relatively complex procedures. Here we developed a visual and rapid loop-mediated isothermal amplification (LAMP) method to amplify three GM rice event-specific junction sequences. Target DNA was amplified and visualized by two indicators (SYBR green or hydroxy naphthol blue [HNB]) within 60 min at an isothermal temperature of 63 °C. Different kinds of plants were selected to ensure the specificity of detection and the results of the non-target samples were negative, indicating that the primer sets for the three GM rice varieties had good levels of specificity. The sensitivity of LAMP, with detection limits at low concentration levels (0.01%-0.005% GM), was 10-
A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay was developed for the rapid detection of equine coronavirus (ECoV). This assay was conducted at 60°C for 40min. Specificity of the RT-LAMP assay was confirmed using several equine intestinal and respiratory pathogens in addition to ECoV. The novel assay failed to cross-react with the other pathogens tested, suggesting i ...
Pathologic lymph node staging is becoming a deficient method in the demanding molecular era. Nevertheless, the use of more sensitive molecular analysis for nodal staging is hampered by its high costs and extensive time requirements. Our aim is to take a step forward in colon cancer (CC) lymph node (LN) pathology diagnosis by proposing a feasible and efficient molecular method in routine practice using reverse transcription loop-mediated isothermal amplification (RT-LAMP). Molecular detection of tumor cytokeratin 19 (CK19) mRNA with RT-LAMP was performed in 3206 LNs from 188 CC patients using two methods: individual analysis of 1449 LNs from 102 patients (individual cohort), and pooled LN analysis of 1757 LNs from 86 patients (pooling cohort). A median of 13 LNs (IQR 10-18) per patient were harvested in the individual cohort, and 18 LNs (IQR 13-25) per patient in the pooling cohort (p ≤ 0.001). The median of molecular assays performed in the pooling cohort was 2 per patient (IQR 1-3), saving a median
Open peer review is a system where authors know who the reviewers are, and the reviewers know who the authors are. If the manuscript is accepted, the named reviewer reports are published alongside the article. Pre-publication versions of the article and author comments to reviewers are available by contacting [email protected] All previous versions of the manuscript and all author responses to the reviewers are also available.. You can find further information about the peer review system here.. ...
Mediators of Inflammation is a peer-reviewed, Open Access journal that publishes original research and review articles on all types of inflammatory mediators, including cytokines, histamine, bradykinin, prostaglandins, leukotrienes, PAF, biological response modifiers and the family of cell adhesion-promoting molecules.
A novel dNAD platform (BEAMing LAMP) by combining emulsion micro-reactors, single-molecule magnetic capture and on-bead loop-mediated isothermal amplification has been developed for DNA detection, which enables absolute and high-precision quantification of a target with a detection limit of 300 copies.
... Gene Amplification technology is one of the rapidly growing markets...San Jose CA (PRWEB) November 4 2008 -- Molecular diagnostics and the...World a href http://www.strategyr.com/Gene_Amplification_Technologi...a href http://www.strategyr.com/Gene_Amplification_Technologies_Mar...,World,Gene,Amplification,Technologies,Market,to,Reach,$2.2,Billion,by,2015,,According,to,New,Report,by,Global,Industry,Analysts,,Inc.,biological,advanced biology technology,biology laboratory technology,biology device technology,latest biology technology
Title: Alabel-free and enzyme-free signal amplification strategy for a sensitive RNase Hactivity assay Author: Chang Yeol Lee,‡Hyowon Jang,‡ Ki Soo Park* and Hyun Gyu Park* (‡: equal contribution) Journal: Nanoscale 2017, 9, 16149-16153 Abstra
Title: Alabel-free and enzyme-free signal amplification strategy for a sensitive RNase Hactivity assay Author: Chang Yeol Lee,‡Hyowon Jang,‡ Ki Soo Park* and Hyun Gyu Park* (‡: equal contribution) Journal: Nanoscale 2017, 9, 16149-16153 Abstra
Background Insulin-like growth factor 1 insensitivity caused by IGF1R mutations has been previously identified as one of the causes of growth impairment in children born small for gestational age (SGA). Objective To analyse the IGF1R in children born SGA. Subjects From an initial cohort of 54 sequential children born SGA, without catch-up growth, 25 children were selected for this IGF1R study due to the presence of serum IGF-1 values above the mean for their age and sex. Methods The proximal IGF1R promoter region, the entire coding region and the exonintron boundaries were directly sequenced, and multiplex ligation-dependent probe amplification analysis was performed. Fibroblast cultures were developed from one patient with a mutation for the in vitro characterization of IGF-1 insensitivity. Results The copy number variation analysis did not identify deletions involving the IGF1R gene. We identified two children carrying heterozygous nucleotide substitutions in IGF1R: c.16G,A/p.Gly6Arg and ...
Finding a gene for familial cardiac fibrosis (NHS grant 2000.130) Summary of results. A large multi-generation family with an autosomal dominantly inherited form of cardiac fibrosis with a highly malignant clinical outcome is presented and used to identify the gene causing this disease. We consider it a hereditary form of cardiac fibrosis since it was shown that the myocardial fibrosis in this family preceded the clinical and echocardiographic signs.. Twenty-four individuals from this family were clinically evaluated and 18 of them were used for a genome wide linkage analysis giving the highest LOD score (2.6) in the region of the lamin AC (LMNA) gene. Mutation analyses of this candidate gene failed to show a point mutation. Subsequent Southern blot and multiplex ligation-dependent probe amplification analyses, however, revealed a deletion of the start-codon containing exon. The up- and downstream flanking exons proved not to be deleted.. Furthermore, we demonstrate in-vitro that the deletion ...
MORSE, Stephen A. Advances in diagnostic tests for bacterial STDs. Salud pública Méx [online]. 2003, vol.45, suppl.5, pp.S698-S708. ISSN 0036-3634.. Because of their asymptomatic nature and nonspecific symptoms, laboratory tests are often required to diagnose a sexually transmitted infection. Over the past few years, there have been advances in technology, such as the development of nucleic acid amplification assays, which have improved our ability to diagnose infections caused by Chlamydia trachomatis. The finding that nucleic acid amplification tests can detect more infected individuals and are useful in screening low prevalence populations, has led to the development of strategies designed to reduce the cost of these assays without significantly impacting their sensitivity. The development of new tests for the diagnosis of syphilis has gained momentum from the report of a synthetic VDRL antigen, which will result in better nontreponemal antibody tests for syphilis. In spite of the ...
Shigella is an important human food-borne zoonosis bacterial pathogen, and can cause clinically severe diarrhea. There is an urgent need to develop a specific, sensitive and rapid methodology for detection of this pathogen. In this study, loop-mediated isothermal amplification (LAMP) combined with magnetic immunocapture assay(IC-LAMP) was first developed for the detection of Shigella in pure culture, artificial milk, and clinical stool samples. This method exhibited a detection limit of 8.7 CFU/mL. Compared with PCR, IC-LAMP is sensitive, specific, and reliable for monitoring Shigella. Additionally, IC-LAMP is more convenient, efficient, and rapid than ordinary LAMP, as it is more efficiently enriches pathogen cells without extraction of genomic DNA. Under isothermal conditions, the amplification curves and the green fluorescence were detected within 30 min in the presence of genomic DNA template. The overall analysis time was approximately 1h, including the enrichment and lysis of the bacterial cells,
GENETIC ANALYSIS LOC FOR NUCLEIC ACID AMPLIFICATION USING NUCLEIC ACID SEQUENCE BASED AMPLIFICATION - diagram, schematic, and image 72 ...
Improved methods for use in nucleic acid amplification, including multiplex amplification, where the amplification is carried out in two or more distinct phases are disclosed. The first phase amplific
The platform - called NINA for non-instrumented nucleic acid amplification - is designed to work with a wide range of isothermal amplification methods.
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Nicking Enzyme Amplification Reaction (NEAR) is a method for in vitro DNA amplification like the polymerase chain reaction (PCR). NEAR is isothermal, replicating DNA at a constant temperature using a polymerase (and nicking enzyme) to exponentially amplify the DNA at a temperature range of 55 °C to 59 °C. One disadvantage of PCR is that it consumes time uncoiling the double-stranded DNA with heat into single strands (a process called denaturation) . This leads to amplification times typically thirty minutes or more for significant production of amplified products.[better source needed] Potential advantages of NEAR over PCR are increased speed and lower energy requirements, characteristics that are shared with other isothermal amplification schemes. A major disadvantage of NEAR relative to PCR is that production of nonspecific amplification products is a common issue with isothermal amplification reactions. The NEAR reaction uses naturally occurring or engineered endonucleases that introduce a ...
Gonorrhea Test NYC. Nucleic Acid Amplification (NAA) test is the most accurate FDA approved test to detect Gonorrhea. It cost only $89.
Reappraising the value of urine leukocyte esterase testing in the age of nucleic acid amplification. The acceptability of oral fluid testing for HIV antibodies: a pilot study in gay bars in a predominantly rural state
A nucleic acid test (NAT) or nucleic acid amplification test (NAAT) is a molecular technique used to detect a particular pathogen (virus or bacterium) in a specimen of blood or other tissue or body fluid. It does so by detecting and amplifying the RNA or DNA of the pathogen, that is, making extra copies of its nucleic acids. This type of medical test was developed to shorten the window period, a time between when a patient has been infected and when they show up as positive by antibody tests such as ELISA. Whereas antibody positivity cannot occur until the immune system has had days or weeks to develop a sizable subpopulation of antibodies specific to the pathogen, a NAT can detect a pathogen as soon as it is present. The term includes any test that directly detects the genetic material of the infecting organism or virus, such as polymerase chain reaction, ligase chain reaction, and others. Their mode of use is usually that a suspected diagnosis is narrowed down on the basis of the history and ...
1 GVRG ABRF 2011 Research Study Evaluation of DNA Whole Genome Amplification (WGA) Technologies for Genotyping The evolution of genomic technologies is occurring rapidly and often requires large amounts of source DNA. There is also an expanded desire to analyze smaller numbers of cells for higher resolution studies as well as to take advantage of large numbers of archived samples (eg. FFPE, serum, etc.). To provide enough material for the newest genomic technologies, whole genome amplification (WGA) has reemerged as an important and necessary technique. With new WGA products on the market, the GVRG has completed a benchmarking study evaluation of 6 commercially available WGA kits using several QC and genotyping assays. Utilizing 6 samples, the WGA kits were tested following the manufacturers protocols. Quality metrics provided measurements on yield, fragment size, and concentration of material derived from each kit. Several widely adopted genotyping methods were then used to evaluate the ...
Researchers often face the problem of limited or insufficient DNA quantity for downstream analysis. QIAGENs whole genome amplification technology solves this problem by providing high yields of whole genomic DNA from small or precious samples, including single cells. REPLI-g Kits and Service accurately copy the original source DNA without bias and the amplified DNA can be directly used in a range of genetic analyses.|br /> |br />
Malaria is one of the leading causes of infectious disease in travelers returning from the tropics. The diagnosis of malaria is typically performed by examining Giemsa-stained thick and thin peripheral blood smears, which is time consuming, labor intensive, and requires high levels of proficiency. Alternatively, loop-mediated isothermal amplification (LAMP) is a new molecular method, which is rapid, sensitive, and requires less capital equipment and technological training. We conducted a retrospective study comparing two formats of a commercial LAMP assay (Meridian illumigene malaria [M] and malaria Plus [MP]) versus reference microscopy on archived blood specimens (n = 140) obtained from unique returning travelers suspected of having malaria ...
Epstein-Barr virus (EBV)-associated disease exhibits distinct gene expression patterns characterized by the transcription of EBV nuclear antigen (EBNA) 1, EBNA2, latent membrane protein (LMP) 1, LMP2A, and BZLF1 (Zebra). A series of visual reverse transcript loop-mediated isothermal amplification (RT-LAMP) assays were performed to examine the expression of EBNA1, EBNA2, LMP1, LMP2A and BZLF1. The sensitivity of RT-LAMP for these transcripts was approximately equivalent to real-time RT-PCR (RT-qPCR), which was developed to quantify relative levels of EBV transcripts, and 10 to 100-fold more sensitive than conventional RT-PCR. Cross-reactions to other viruses were not observed upon examination of cell lines infected with herpes simplex viruses-1 and -2 (HSV-1 and -2), varicella zoster virus (VZV), human cytomegalovirus (HCMV) or Kaposis sarcoma-associated herpesvirus. When applied to 146 specimens, RT-LAMP exhibited high clinical sensitivity and specificity, with an excellent agreement (κ > 0.92)
RNA amplification is necessary for profiling gene expression from small tissue samples. Previous studies have shown that the T7 based amplification techniques are reproducible but may distort the true abundance of targets. However, the consequences of such distortions on the ability to detect biological variation in expression have not been explored sufficiently to define the true extent of usability and limitations of such amplification techniques. We show that expression ratios are occasionally distorted by amplification using the Affymetrix small sample protocol version 2 due to a disproportional shift in intensity across biological samples. This occurs when a shift in one sample cannot be reflected in the other sample because the intensity would lie outside the dynamic range of the scanner. Interestingly, such distortions most commonly result in smaller ratios with the consequence of reducing the statistical significance of the ratios. This becomes more critical for less pronounced ratios where the
A collection of functions to pre-process amplification curve data from polymerase chain reaction (PCR) or isothermal amplification reactions. Contains functions to normalize and baseline amplification curves, to detect both the start and end of an amplification reaction, several smoothers (e.g., LOWESS, moving average, cubic splines, Savitzky-Golay), a function to detect false positive amplification reactions and a function to determine the amplification efficiency. Quantification point (Cq) methods include the first (FDM) and second approximate derivative maximum (SDM) methods (calculated by a 5-point-stencil) and the cycle threshold method. Data sets of experimental nucleic acid amplification systems (VideoScan HCU, capillary convective PCR (ccPCR)) and commercial systems are included. Amplification curves were generated by helicase dependent amplification (HDA), ccPCR or PCR. As detection system intercalating dyes (EvaGreen, SYBR Green) and hydrolysis probes (TaqMan) were used. ...
A collection of functions to pre-process amplification curve data from polymerase chain reaction (PCR) or isothermal amplification reactions. Contains functions to normalize and baseline amplification curves, to detect both the start and end of an amplification reaction, several smoothers (e.g., LOWESS, moving average, cubic splines, Savitzky-Golay), a function to detect false positive amplification reactions and a function to determine the amplification efficiency. Quantification point (Cq) methods include the first (FDM) and second approximate derivative maximum (SDM) methods (calculated by a 5-point-stencil) and the cycle threshold method. Data sets of experimental nucleic acid amplification systems (VideoScan HCU, capillary convective PCR (ccPCR)) and commercial systems are included. Amplification curves were generated by helicase dependent amplification (HDA), ccPCR or PCR. As detection system intercalating dyes (EvaGreen, SYBR Green) and hydrolysis probes (TaqMan) were used. ...
In this study, we were able to demonstrate that the performance of the TRC method for the detection of MTC from respiratory samples was comparable to that of the COBAS AMPLICOR PCR in terms of sensitivity and specificity. The TRC method detected as little as one BCG organism per 100 μl of sputum. The selection of rRNA as the amplification target, of which a single cell contains multiple copies, may be the primary reason for this favorable result.. In the case of MTC smear-negative and culture-positive samples, the TRC method showed substantially lower rates of positivity than it did for smear- and culture-positive samples, as did the COBAS AMPLICOR PCR. This less favorable result was, however, comparable to the results of previous studies that evaluated nucleic acid amplification assays in similar settings (18). As for the specificity, the TRC method did not show positivity for 106M. avium or 106M. kansasii organisms per 100 μl of sputum, nor did it show positivity for 106 initial copies of ...
OCP ligation/ERCA can be used to directly probe genomic DNA for SNPs or mutations, and the entire process can be performed in a single tube. The ERCA amplification reaction is rapid, generating signal in as little as 10 minutes, and is incubated at a single temperature. These characteristics make OCP ligation/ERCA easily adaptable to high throughput automated genotyping platforms. Accurate genotyping was obtained in screens designed to detect four clinically relevant mutations, Factor V Leiden, Factor II prothrombin, Hemochromatosis C282Y and Hemochromatosis H63D.. Several improvements to the ligation/ERCA method [8] have increased accuracy to levels acceptable for diagnostic applications and reduced reaction time. Other reports using ERCA based genotyping require PCR amplification of the locus of interest prior to genotyping [3, 16], which is prohibitively expensive, time consuming, and more difficult to automate. We have designed ERCA primers that are optimized for minimal misamplification and ...
Sigmas GenomePlex Whole Genome Amplification (WGA) kit utilizes a proprietary amplification method designed for robust and accurate amplification of limited source DNA. In less than three hours, GenomePlex WGA successfully amplifies nanogram amounts of starting DNA, regardless of source, into microgram yields.
Scientists at the University of Pennsylvania have developed a point-of-care device for detecting the Zika virus for under $2 a pop. Its about the size of a can of coke and can be operated by just about anyone, and even outside the clinic. A patient simply spits into the receptacle and after a while a paper test strip changes its color to a shade of blue if a Zika virus is detected.. Previously available tests either take a lot of time, produce too many false negatives, or require expensive equipment that limit their usability. RT-PCR, particularly, works well but requires precise sample management to amplify the DNA.. The new test uses a technique called loop-mediated isothermal amplification (LAMP) to grow the viral DNA. One problem of applying LAMP in the past has been developing so-called "primers," short DNA strings that complement the DNA of the viral target. The researchers used computational techniques to identify spots on the Zika DNA that are unique to that virus, which were integrated ...
Objective: To analyze the pathogens of lower respiratory tract infection(LRTI) including bacterial, viral and mixed infection, and to establish a discriminant model based on clinical features in order to predict the pathogens. Methods: A total of 243 hospitalized patients with lower respiratory tract infections were enrolled in Fujian Provincial Hospital from April 2012 to September 2015. The clinical data and airway (sputum and/or bronchoalveolar lavage) samples were collected. Microbes were identified by traditional culture (for bacteria), loop-mediated isothermal amplification(LAMP) and gene sequencing (for bacteria and atypical pathogen), or Real-time quantitative polymerase chain reaction (Real-time PCR)for viruses ...
... : A single cell whole genome amplification kit based on a revolutionary multiple displacement amplification method.
A method for determining an unknown starting quantity of a target nucleic acid sequence in a test sample comprises amplifying the unknown starting quantity of the target nucleic acid sequence in the test sample and known starting quantities of a calibration nucleic acid sequence in respective calibration samples; and determining a respective threshold value for each of the nucleic acid sequences using a derivative of a growth curve derived for the sequence. The starting quantity of the target nucleic acid sequence in the test sample is determined using the threshold value determined for the target sequence and a calibration curve derived from the threshold values determined for the known starting quantities of the calibration nucleic acid sequences.
Whole Genome Amplification (WGA) provides the means to immortalize genomic DNA from fresh, frozen, archived and chemically treated samples. GenomePlex® WGA, produces 500-1000 fold unbiased amplification of genomic DNA via a PCR-based sequence independent approach.
Disease marker detection plays an important role in clinical practice; however, rapid, low-cost and simultaneous detection of multiple trace disease markers remain a challenge because of low abundance of related strategies. Herein, we report a G-quadruplex DNAzyme and nicking enzyme-assisted multiplex chemil
The data from our study are consistent with other publications that demonstrate the increased sensitivity of molecular detection methods for HSV compared to that of cell culture. This is illustrated by the large number of specimens testing positive by illumigene but negative by ELVIS. To support the presence of HSV nucleic acid in this cohort, 83 specimens with sufficient volume for additional analysis were tested using another isothermal amplification assay based on a different HSV genetic target (AmpliVue HSV1+2; Quidel). The AmpliVue result was in agreement with illumigene for 57 of 83 (78.1%) specimens, generating initial discrepant results between illumigene and ELVIS. The remaining eight specimens were unavailable for discrepant analysis (7 mucocutaneous and 1 cutaneous). A limitation of the discrepant analysis was that all specimens were frozen prior to AmpliVue testing. This may have reduced the sensitivity of AmpliVue since freeze-thaw cycles may reduce sensitivity of molecular assays. ...
|p|Whole genome amplification was developed in 1992 (1, 2) as a way of increasing the amount of limited DNA samples. This is particularly useful for forensics and genetic disease research, where DNA quantities are limited, but many analyses are required. Various WGA techniques have been developed that differ both in their protocols, amplification accuracy, and ease-of-use.|/p|
Viruses strongly influence microbial population dynamics and ecosystem functions. However, our ability to quantitatively evaluate those viral impacts is limited to the few cultivated viruses and double-stranded DNA (dsDNA) viral genomes captured in quantitative viral metagenomes (viromes). This leaves the ecology of non-dsDNA viruses nearly unknown, including single-stranded DNA (ssDNA) viruses that have been frequently observed in viromes, but not quantified due to amplification biases in sequencing library preparations (Multiple Displacement Amplification, Linker Amplification or Tagmentation).Here we designed mock viral communities including both ssDNA and dsDNA viruses to evaluate the capability of a sequencing library preparation approach including an Adaptase step prior to Linker Amplification for quantitative amplification of both dsDNA and ssDNA templates. We then surveyed aquatic samples to provide first estimates of the abundance of ssDNA viruses.Mock community experiments confirmed ...
The accuracy of the Gen-Probe APTIMA system in published studies is in the range of 98-100%, but we performed our own validation of the APTIMA system to insure good performance for testing of BC patients. Cooperating clinics collected dual Gen-Probe APTIMA and BD ProbeTec swabs that were tested in parallel on the two systems. We also tested urine specimens on the two systems. Discrepancies were resolved by repeat testing on both systems and by testing using alternative DNA amplification methods. 1534 specimens were tested by both methods for Chlamydia and gonorrhea. A total of 269 specimens were positive for one or both organisms, and the APTIMA system demonstrated ,99% sensitivity and specificity overall for both swab and urine specimens compared to the BD ProbeTec system. The results by specimen type are shown above. Based on this excellent performance, the Gen-Probe APTIMA system was chosen for implementation.. Learn more details ...
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Signal-mediated amplification of RNA technology (SMART) is a novel isothermal amplification technology that uses a three-way junction (3WJ) structure to
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AgNCs are complexes between few Ag atoms and a specific DNA sequence template to stabilize the clusters. In most cases, AgNCs are synthesized either at the 3 or 5 end of the template. Our results show that AgNCs can successfully be generated from a template embedded in the middle of a hybridization probe, used for the isothermal amplification of aptamers, called rolling circle amplification (RCA). RCA uses circular oligonucleotide probes to generate long, ssDNA molecules containing periodic repeats of the circular probe.[4][5] Previous works show that overexpression of aptamers by RCA increases target binding efficiency compared to monovalent aptamers.[6] The RCA concatemer combines both the aptamer and the fluorescent AgNC template. Subsequently synthetized AgNCs exhibit strong, robust and tunable fluorescence, eliminating the need for labeling.[7] Importantly, it has been shown that aptamer-AgNCs retain the same specificity and affinity for the cognate protein and that target binding results ...
... Improve PCR product yield and specificity for difficult amplificati...Certain PCR amplification systems produce multiple product bands or...Stratagene is committed to developing innovative high-performance PCR...,Optimizing,pfuturbo,DNA,Polymerase,Amplification,Reactions,with,,,Perfect,Match,PCR,Enhancer,biological,advanced biology technology,biology laboratory technology,biology device technology,latest biology technology
The report also highlights country successes - among them Cambodia which has seen a 45% drop in TB prevalence between 2002 and 2011 - and, in all, it features data from 204 countries and territories and covers all aspects of TB, including multidrug-resistant TB (MDR-TB), TB/HIV, research and development (R&D) and TB financing.. There is praise in the report for the worldwide roll-out of a new diagnostic device that can test patients for TB, including drug-resistant TB, in just 100 minutes. The fully automated nucleic acid amplification test (NAAT), which can diagnose TB and rifampicin-resistant disease, is now available in 67 low- and middle-income countries. Adoption of the while you wait test is expected to further accelerate following a recent 41% fall in the price of the test.. The report also points to the promise of medical breakthroughs from new TB drugs - the first in over 40 years - which could be on the market as early as 2013. Indeed, tools to prevent, detect and treat all forms of ...
Management of tuberculosis (TB) is challenging in HIV/TB-coinfected children. The World Health Organization recommends nucleic acid amplification tests for TB diagnosis, a 4-drug regimen including ethambutol during intensive phase (IP) of treatment, and initiation of antiretroviral therapy (ART) wit...
Since the discovery of the doublehelix structure of DNA (1), no single event has had the same impact on the field of molecular biology as the rediscovery by Kary ...
The safety and scientific validity of this study is the responsibility of the study sponsor and investigators. Listing a study does not mean it has been evaluated by the U.S. Federal Government. Read our disclaimer for details ...
SELECTIVE AMPLIFICATION OF OVERLAPPING AMPLICONS | METHODS AND COMPOSITIONS FOR LABELING TARGETS | DNA DETECTION METHOD AND DNA DETECTION DEVICE | ROLLING CIRCLE AMPLIFICATION METHOD | METHOD AND DEVICE FOR DETECTING MOLECULES OR PARTICLES USING FRACTIONALIZED VOLUMES |
In this project we will develop kits, services, and reagents useful in the processes of generating and employing amplified DNA mini-libraries (i.e., amplified products) from microdissected human chromosome fragments. The focus of Phase I studies will be development of standard protocols for generating such mini-libraries. Two amplification strategies are proposed based on their use of recognition sites distributed throughout the human genome at about 250 bp intervals (insuring representative coverage of genomic sequences in the min- library). The strategies are based on DNA sequence recognition by Mbo I cleavage and by oligonucleotide hybridization to the most common 6 bp sequences in the human genome. DNA products obtained from both strategies are PCR amplified with a single adaptor oligonucleotide. The products of the amplification reactions will be analyzed for the amount and size of DNA, degree of genome coverage, hybridization specificity, retrievability upon reamplification, ease of use ...
Because only one primer is used, only one strand is copied during sequencing, there is a linear (not log as in PCR) increase of the number of copies of one strand of the gene. Therefore, there has to be a large number of copies of the gene in the starting mixture for sequencing. If there are 1000 copies of the wanted gene before the cycling starts, after one cycle there will be 2000 copies: the 1000 original templates and 1000 complementary strands with each one fluorescent label on the last base. After two cycles, there will be 2000 complementary strands, three cycles will result in 3000 complementary strands and so on. Linear amplification during sequencing reactions explains why we have to start with an exponential amplification of the gene of interest in a polymerase chain reaction to get lots of copies of our gene BEFORE we do the sequencing. Both sequencing and polymerase chain reactions require short sequences of DNA called primers to direct the specificity of the amplification to only ...
Because only one primer is used, only one strand is copied during sequencing, there is a linear (not log as in PCR) increase of the number of copies of one strand of the gene. Therefore, there has to be a large number of copies of the gene in the starting mixture for sequencing. If there are 1000 copies of the wanted gene before the cycling starts, after one cycle there will be 2000 copies: the 1000 original templates and 1000 complementary strands with each one fluorescent label on the last base. After two cycles, there will be 2000 complementary strands, three cycles will result in 3000 complementary strands and so on. Linear amplification during sequencing reactions explains why we have to start with an exponential amplification of the gene of interest in a polymerase chain reaction to get lots of copies of our gene BEFORE we do the sequencing. Both sequencing and polymerase chain reactions require short sequences of DNA called primers to direct the specificity of the amplification to only ...
Unlike end point PCR (conventional PCR) real time PCR allows quantification of the desired product at any point in the amplification process by measuring fluorescence (in reality, measurement is made of its level over a given threshold). A commonly employed method of DNA quantification by quantitative PCR relies on plotting fluorescence against the number of cycles. A threshold for detection of DNA-based fluorescence is set slightly above background. The number of cycles at which the fluorescence exceeds the threshold is called the threshold cycle (Ct) or, according to the MIQE guidelines, quantification cycle (Cq). During the exponential amplification phase, the quantity of the target DNA template (amplicon) doubles every cycle. For example, a DNA sample whose Cq precedes that of another sample by 3 cycles contained 23 = 8 times more template. However, the efficiency of amplification is often variable among primers and templates. Therefore, the efficiency of a primer-template combination is ...
As the worlds agricultural systems endeavour to sustain an expanding population, technologies have become available to increase the yield and viability of cultivated crops including the introduction of novel traits into crops using genetic transformation of foreign DNA to produce GM varieties. However, public resistance to commercialization of genetically modified plants is still widespread in Europe [1, 2]. Existing European regulation limits the extent of GM presence in non-GM foodstuffs, and the increasing introduction of GM products into Europe is likely to result in parallel GM and non-GM ("conventional") supply chains. In addition, the more widespread planting of GM crops in Europe will lead to the need for on-farm confirmation of GM status. Together these factors are likely to lead to a substantial increase in the extent and frequency of testing for the presence of DNA of a GM-derived origin.. The European Union has currently defined the proportion of GM that can be present to be no more ...
The goal of this project, over Phases I and II, is to create a new, accurate and quantitative platform for massively parallel, ultra high throughput screening of SNPs, mutations, and gene expression on solid supports, e.g. chips. This platform, based on our novel, linear signal amplification technology, the Invader assay, will immediately impact the effort to associate SNPs and mutations with phenotypes and will be of increasing value in the molecular profiling of cancer. High throughput SNP analysis is an active area of technology development; nonetheless, most existing technologies are hampered by exclusive reliance on allele specific hybridization. The Invader assay is based on a highly specific enzyme-substrate reaction (i.e. cleavage of a precise structure formed by oligonucleotides hybridized to a target sequence). The unique accuracy of this method results from the combined specificity conferred by this sequence-specific probe hybridization and structure-specific enzymatic cleavage. This ...
This thesis describes a new approach to biomolecular analysis, called the volume-amplified magnetic nanobead detection assay (VAM-DNA). It is a sensitive, specific magnetic bioassay that offers a potential platform for the development of low-cost, easy-to-use diagnostic devices. The VAM-NDA consists of three basic steps: biomolecular target recognition, enzymatic amplification of the probe-target complex using the rolling circle amplification (RCA) technique, and addition of target complementary probe-tagged magnetic nanobeads which exhibit Brownian relaxation behavior. Target detection is demonstrated by measuring the frequency-dependent complex magnetization of the magnetic beads. The binding of the RCA products (target DNA-sequence coils) to the bead surface causes a dramatic increase in the bead size, corresponding essentially to the size of the DNA coil (typically around one micrometer). This causes a decrease in the Brownian relaxation frequency, since it is inversely proportional to the ...
Sygnis TruePrime™ Whole Genome Amplification is a whole genome amplification kit to uniformly amplifies genomic DNA from purified starting material with less bias.
I added also general information (mostly from the Qiagen site) about the REPLI-g method of amplification and about other similar Qiagen kits available ...
Aureon Biosystems is a specialist biotechnology company with three core areas of expertise; magnetic particle based sample preparation systems (A-BeadsTM), nucleic acid amplification systems and chemiluminescent based diagnostic technology ...
Combustion systems with advanced injection strategies have been extensively studied, but there still exists a significant fundamental knowledge gap on fuel spray interactions with the piston surface and chamber walls. This paper is meant to provide detailed data on spray-wall impingement physics and support the spray-wall model development. The experimental work of spray-wall impingement with non-vaporizing spray characterization, was carried out in a high pressure-temperature constant-volume combustion vessel. The simultaneous Mie scattering of liquid spray and schlieren of liquid and vapor spray were carried out. Diesel fuel was injected at a pressure of 1500 bar into ambient gas at a density of 22.8 kg/m3 with isothermal conditions (fuel, ambient, and plate temperatures of 423 K). A Lagrangian-Eulerian modeling approach was employed to characterize the spray-gas and spray-wall interactions in the CONVERGETM framework by means of a Reynolds-Averaged Navier-Stokes (RANS) formulation ...
The curing reaction of a conductive adhesive was studied with a differential scanning calorimeter (DSC) under isothermal conditions in the range of 100-160
Electropheris , Nucleic Acid Purification , PCR - Nucleic Acid Amplification , Nucleic Acid Quantification , Cloning & Restriction Enzyme , DNA Sequencing , Blotting , Transfections , Microarrays , Nucleic Acid Labelling , ...
Based on nucleic acid amplification, Speed-oligo® presents all the advantages of molecular biology in a quick detection for infectious diseases multiplex testing. This PCR-based method coupled to a strip device enables an immediate, highly sensitive and specific detection ...
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