scholarly journals RT-qPCR Testing of SARS-CoV-2: A Primer

2020 ◽  
Vol 21 (8) ◽  
pp. 3004 ◽  
Author(s):  
Stephen A. Bustin ◽  
Tania Nolan
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Diagnostic Tool ◽  
Acute Infection ◽  
Chain Reaction ◽  
Reliable Test ◽  
Polymerase Chain ◽  
Quantitative Fluorescence

Testing for the presence of coronavirus is an essential diagnostic tool for monitoring and managing the current COVID-19 pandemic. The only reliable test in current use for testing acute infection targets the genome of SARS-CoV-2, and the most widely used method is quantitative fluorescence-based reverse transcription polymerase chain reaction (RT-qPCR). Despite its ubiquity, there is a significant amount of uncertainty about how this test works, potential throughput and reliability. This has resulted in widespread misrepresentation of the problems faced using this test during the current COVID-19 epidemic. This primer provides simple, straightforward and impartial information about RT-qPCR.

scholarly journals Evaluation of the Sensitivity of Reverse-Transcription Polymerase Chain Reaction to Detect Porcine Reproductive and Respiratory Syndrome Virus on Individual and Pooled Samples from Boars

2007 ◽  
Vol 19 (5) ◽  
pp. 502-509 ◽  
Author(s):  
Albert Rovira ◽  
Travis Clement ◽  
Jane Christopher-Hennings ◽  
Bob Thompson ◽  
Mark Engle ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Biological Samples ◽  
Acute Infection ◽  
Respiratory Syndrome Virus ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
The Impact ◽  
Pooled Samples

Boar studs are continuously monitored for the presence of porcine reproductive and respiratory syndrome virus (PRRSV) by testing different biological samples by reverse-transcription polymerase chain reaction (RT-PCR). In most cases, samples are run in pools, even though the impact of pooling on the sensitivity of RT-PCR is unknown. The objective of this study was to evaluate the feasibility of using PCR on pooled samples through the estimation of the sensitivity of RT-PCR on different biological samples run individually, in pools of 3 and in pools of 5. Twenty-nine boars were inoculated with a low virulent PRRSV isolate. Serum, blood swab, and semen samples were obtained from each boar every 2 to 3 days for 2 weeks. Each sample was tested by RT-PCR undiluted or diluted 1:3 and 1:5 with negative samples. Eleven of the 29 boars did not appear to get infected from the inoculum, as evidenced by no seroconversion 15 days after inoculation. Data from the other 18 boars showed that serum was the best sample to detect PRRSV during acute infection, with the blood swab sample performing almost as well. Semen samples failed to detect PRRSV infection in most of the cases. Pooling samples at pool sizes of 3 and 5 resulted in a decrease in the sensitivity of RT-PCR. Sensitivity was reduced by 6% and 8%, respectively, when serum or blood swab samples were run in pools of 5. The impact of pooling on the sensitivity of PCR was higher in samples taken during the beginning of the viremic period.

1504: Molecular Diagnosis and Staging of Prostate Cancer Using Clusterin in Real Time Reverse Transcription Polymerase Chain Reaction (RT-PCR)

2006 ◽  
Vol 175 (4S) ◽  
pp. 485-486
Author(s):  
Sabarinath B. Nair ◽  
Christodoulos Pipinikas ◽  
Roger Kirby ◽  
Nick Carter ◽  
Christiane Fenske
Keyword(s):  
Prostate Cancer ◽  
Polymerase Chain Reaction ◽  
Real Time ◽  
Molecular Diagnosis ◽  
Reverse Transcription ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain

Expression of hyaluronan synthase-3 in porcine oviducal epithelium during oestrus

2003 ◽  
Vol 15 (2) ◽  
pp. 99 ◽  
Author(s):  
Paisan Tienthai ◽  
Naoko Kimura ◽  
Paraskevi Heldin ◽  
Eimei Sato ◽  
Heriberto Rodriguez-Martinez
Keyword(s):  
Polymerase Chain Reaction ◽  
Reverse Transcription ◽  
Hyaluronan Synthase ◽  
Critical Periods ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Mean Values ◽  
Polymerase Chain ◽  
Sperm Reservoir ◽  
Oviduct Fluid

Hyaluronan (HA) has been related to fertilization and embryo development in the pig. Furthermore, HA is present in pig oviduct fluid and the lining epithelium, particularly of the pre-ovulatory sperm reservoir. Because the mechanisms that regulate HA synthesis have not yet been clarified, semiquantitative reverse transcription-polymerase chain reaction (RT-PCR) was conducted to assess the expression of mRNAs of two HA-synthesizing enzymes (has2 and has3) in the oviduct epithelium (uterotubal junction, isthmus, ampullary–isthmic junction and ampulla segments) of non-inseminated (control) and inseminated (treatment) sows at pre-, peri- and post-ovulatory oestrus. Only has3 mRNA was detected; it was present in all tubal segments of both control and treatment samples. The level of has3 expression did not vary significantly between non-inseminated and inseminated specimens, but there was a tendency (NS) for increased mean values during the peri- and post-ovulatory stages compared with pre-ovulation. It is concluded that has3 is expressed by the porcine endosalpinx epithelium and the levels of expression do not vary during the critical periods of sperm transport and fertilization, despite fluctuating levels of HA in the tubal fluid at corresponding periods.

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