Two-step RT-PCR
Development and evaluation of one step single tube multiplex RT PCR for rapid detection and typing of dengue viruses
... multiplex RT-PCR and two step RT-PCR followed by nested PCR, which revealed 100% concordance (Details not ...multiplex RT-PCR assay was compared with closely ...
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Optimization of a Combined Human Parechovirus Enterovirus Real Time Reverse Transcription PCR Assay and Evaluation of a New Parechovirus 3 Specific Assay for Cerebrospinal Fluid Specimen Testing
... HPeV PCR methods have been published (16–19), the assays were not properly validated either with sufficient numbers of clinical specimens or with controls for clinical ...and two-step reaction ...
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Standardization of one step single tube rt pcr for rapid detection of all circulating strains of dengue viruses
... ELISA, RT-PCR etc. RT- PCR assay is more sensitive than virus isolation, Virus isolation is although gold standard but time ...time PCR assay is most rapid, sensitive and specific but ...
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Fast Duplex One Step Reverse Transcriptase PCR for Rapid Differential Detection of West Nile and Japanese Encephalitis Viruses
... transcriptase PCR (RT-PCR) methods have been used to develop sensitive and specific assays for the identification of WNV (13, ...(TaqMan) PCR, SYBR green-based real-time PCR, and loop- ...
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Rapid Simultaneous Detection of Enterovirus and Parechovirus RNAs in Clinical Samples by One Step Real Time Reverse Transcription PCR Assay
... as PCR, has substantially improved the sensitivity of enterovirus detection compared to that of virus culture ...methods. PCR-based methods also can detect a much wider range of EV variants, including those ...
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Detection of a novel avian influenza A (H7N9) virus in humans by multiplex one step real time RT PCR assay
... these two indeterminate tissue culture results were subsequently confirmed to be positive by genomic sequence analysis and were excluded from the sensitivity and specificity ...by PCR assay were repeated at ...
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Development of one step SYBR Green real time RT PCR for quantifying bovine viral diarrhea virus type 1 and its comparison with conventional RT PCR
... nested PCR [13] for the detection of BVDV have been ...[14]. RT-PCR is sensi- tive and specific for the detection of BVDV, but the ana- lysis of RT-PCR amplified fragment is usually ...
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A one step real time RT PCR assay for the quantitation of Wheat yellow mosaic virus(WYMV)
... of RT-qPCR, however, re- quires the use of reference genes whose level of expres- sion is not affected by the test, general physiological conditions or inter-individual ...a RT-qPCR assay that can be used ...
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Superiority of Digital Reverse Transcription PCR (RT PCR) over Real Time RT PCR for Quantitation of Highly Divergent Human Rhinoviruses
... conventional PCR using the type-specific T7 polymerase promoter-labeled forward primer and type-specific reverse ...for RT-qPCR and 1e2 to 1e6 copies/reaction for RT-dPCR) were amplified using the ...
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One-Step RT-qPCR assay for detection and quantification of equine infectious anemia virus in-vitro
... One Step qRT-PCR system was performed in ...competitive RT-PCR and analysis of gel electrophoresis stained with ethidium bromide generated a minimum detection limit of 100 molecules with a ...
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Design and Validation of an H5 TaqMan Real Time One Step Reverse Transcription PCR and Confirmatory Assays for Diagnosis and Verification of Influenza A Virus H5 Infections in Humans
... real-time RT-PCR assay was shown to be extremely sensitive for the detection of influenza A H5 viruses, detecting clade 1, 2, and 3 H5 virus levels of 1 PFU or less by using a QuantiTect Probe ...
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Specific detection of H5N1 avian influenza A virus in field specimens by a one step RT PCR assay
... In a separate study, the sensitivity of this H5N1 primer set was also evaluated against the currently recommended World Health Organization (WHO) H5 primer set. Three different strains were used – one human archived RNA ...
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Development and evaluation of a real time one step Reverse Transcriptase PCR for quantitation of Chandipura Virus
... real-time RT-PCR assay was performed using the real- time one step RT-PCR master mix (Applied Biosystems International, Foster City, ...multiScribe RT and RNase inhibitor mix, ...
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New quick method for isolating RNA from laser captured cells stained by immunofluorescent immunohistochemistry; RNA suitable for direct use in fluorogenic TaqMan one-step real-time RT-PCR
... each PCR to proceed with optimal efficiency. To accomplish this, two separate optimization plates are set up for each target; the second of which can only be set up after results from the first plate are ...
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Molecular Detection of Hepatitis C Virus (HCV) by Conventional One-step RT-PCR Coupled with Nested PCR
... one-step RT-PCR assay followed by nested ...on two rounds of PCR amplification thereby increasing the detection time but the sensitivity and specificity of the assay was ...of ...
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Development and Validation of a Quantitative PCR Assay for Diagnosis of Scedosporiosis
... first step for preincubation (activation of the enzyme) and denaturation of the template DNA at 95°C for 10 ...prolificans. PCR products were subjected to electrophoresis in 2% agarose gels (Prona- disa, ...
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Highly Sensitive Assay for Detection of Enterovirus in Clinical Specimens by Reverse Transcription PCR with an Armored RNA Internal Control
... the RT-PCR was evaluated with limiting dilutions of both armored EV RNA and armored IC ...by PCR with a single copy of DNA ...by PCR should represent 1 copy of IC ...efficient RT ...
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Evidence that the intra amoebal Legionella drancourtii acquired a sterol reductase gene from eukaryotes
... homologs, two plausible hypotheses were inferred: i) the ATP/ADP translocase was acquired from a plant by the Chlamydia ancestor that might have been a plant parasite and was transferred to rickettsiae [21] or ii) ...
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Identification of Genetic Evidence for Dobrava Virus Spillover in Rodents by Nested Reverse Transcription (RT) PCR and TaqMan RT PCR
... the two rodent host species overlaps in Europe, but rodent carriers of both DOBV types coexist only in the border region of southeastern and central Europe (2, ...
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Heminested reverse transcriptase polymerase chain reaction (hnRT PCR) as a tool for rabies virus detection in stored and decomposed samples
... Background: The use of methods, both sensitive and specific, for rabies diagnosis are important tools for the control and prophylaxis of the disease. Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR) ...
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