scholarly journals Development and validation of a reverse transcription quantitative PCR for universal detection of viral hemorrhagic septicemia virus

2011 ◽  
Vol 95 (2) ◽  
pp. 97-112 ◽  
Author(s):  
KA Garver ◽  
LM Hawley ◽  
CA McClure ◽  
T Schroeder ◽  
S Aldous ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
Reverse Transcription ◽  
Viral Hemorrhagic Septicemia Virus ◽  
Viral Hemorrhagic Septicemia ◽  
Reverse Transcription Quantitative Pcr ◽  
Universal Detection ◽  
Development And Validation

scholarly journals Pan-Piscine Orthoreovirus (PRV) Detection Using Reverse Transcription Quantitative PCR

Pathogens ◽  
2021 ◽  
Vol 10 (12) ◽  
pp. 1548
Author(s):  
Julie Zhao ◽  
Niccolò Vendramin ◽  
Argelia Cuenca ◽  
Mark Polinski ◽  
Laura M. Hawley ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
Reverse Transcription ◽  
High Sensitivity ◽  
Test Results ◽  
Reverse Transcription Quantitative Pcr ◽  
Highly Sensitive ◽  
Surveillance Programs ◽  
America South ◽  
Universal Detection ◽  
Quantitative Pcr Assay

Piscine orthoreovirus (PRV) infects farmed and wild salmon and trout species in North America, South America, Europe, and East Asia. PRV groups into three distinct genotypes (PRV-1, PRV-2, and PRV-3) that can vary in distribution, host specificity, and/or disease potential. Detection of the virus is currently restricted to genotype specific assays such that surveillance programs require the use of three assays to ensure universal detection of PRV. Consequently, herein, we developed, optimized, and validated a real-time reverse transcription quantitative PCR assay (RT-qPCR) that can detect all known PRV genotypes with high sensitivity and specificity. Targeting a conserved region at the 5′ terminus of the M2 segment, the pan-PRV assay reliably detected all PRV genotypes with as few as five copies of RNA. The assay exclusively amplifies PRV and does not cross-react with other salmonid viruses or salmonid host genomes and can be performed as either a one- or two-step RT-qPCR. The assay is highly reproducible and robust, showing 100% agreement in test results from an inter-laboratory comparison between two laboratories in two countries. Overall, as the assay provides a single test to achieve highly sensitive pan-specific PRV detection, it is suitable for research, diagnostic, and surveillance purposes.

scholarly journals Establishment and Application of Rapid Diagnosis for Reverse Transcription-Quantitative PCR of Newly Emerging Goose-Origin Nephrotic Astrovirus in China

mSphere ◽  
2018 ◽  
Vol 3 (6) ◽  
Author(s):  
Xiaoyuan Yuan ◽  
Kai Meng ◽  
Yuxia Zhang ◽  
Lihong Qi ◽  
Wu Ai ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
Clinical Application ◽  
Reverse Transcription ◽  
Test Sample ◽  
Sensitivity Testing ◽  
Content Type ◽  
Reverse Transcription Quantitative Pcr ◽  
Emerging Virus ◽  
New Type ◽  
And Control

ABSTRACT In 2017, a new type of goose-origin astrovirus (GoAstV) that is completely different from previously identified avian astroviruses (which have only 30.0% to 50.5% homology with GoAstV) has been isolated from diseased geese in China. This disease can cause joint swelling in sick geese, and the anatomy shows a clear precipitation of urate in the kidney. The rate of death and culling can reach more than 30%, revealing the disease’s severe pathogenicity. To quickly and accurately diagnose the newly emerging disease, we established a highly specific reverse transcription-quantitative PCR (RT-qPCR) method of detecting GoAstV. Sensitivity testing showed that the minimum amount of test sample for this method is 52.5 copies/μl. Clinical application confirmed that this method can quickly and effectively detect GoAstV, providing a diagnostic platform for the prevention and control of goose disease. IMPORTANCE Goose-origin astrovirus (GoAstV), as a newly emerging virus in 2017, is different from previously known astroviruses in the genus Avastrovirus. So far, few studies have focused on the novel virus. Considering the infectious development of astrovirus (AstV), we established a reverse transcription-quantitative PCR (RT-qPCR) assay with a strong specificity to quickly and accurately diagnose GoAstV. Confirmed by clinical application, this method can quickly and accurately detect prevalent GoAstV. The assay is thus convenient for clinical operation and is applicable to the monitoring of GoAstV disease.

scholarly journals Selection of Valid Reference Genes for Reverse Transcription Quantitative PCR Analysis inHeliconius numata(Lepidoptera: Nymphalidae)

2016 ◽  
Vol 16 (1) ◽  
pp. 50 ◽  
Author(s):  
Florence Piron Prunier ◽  
Mathieu Chouteau ◽  
Annabel Whibley ◽  
Mathieu Joron ◽  
Violaine Llaurens
Keyword(s):  
Quantitative Pcr ◽  
Reverse Transcription ◽  
Reference Genes ◽  
Pcr Analysis ◽  
Reverse Transcription Quantitative Pcr ◽  
Selection Of

scholarly journals Diversity of Intestinal Clostridium coccoides Group in the Japanese Population, as Demonstrated by Reverse Transcription-Quantitative PCR

PLoS ONE ◽  
2015 ◽  
Vol 10 (5) ◽  
pp. e0126226 ◽  
Author(s):  
Takashi Kurakawa ◽  
Kiyohito Ogata ◽  
Kazunori Matsuda ◽  
Hirokazu Tsuji ◽  
Hiroyuki Kubota ◽  
...  
Keyword(s):  
Quantitative Pcr ◽  
Reverse Transcription ◽  
Japanese Population ◽  
Reverse Transcription Quantitative Pcr
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