Dual priming oligonucleotide (DPO)-based real-time RT-PCR assay for accurate differentiation of four major viruses causing porcine viral diarrhea

Publication date: Available online 12 August 2019Source: Molecular and Cellular ProbesAuthor(s): Shuo Jia, Baohua Feng, Zhuo Wang, Yingying Ma, Xuwen Gao, Yanping Jiang, Wen Cui, Xinyuan Qiao, Lijie Tang, Yijing Li, Li Wang, Yigang XuAbstractCurrently in China, porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine rotavirus (PoRV), and porcine deltacoronavirus (PDCoV) are the major causes of porcine viral diarrhea, and mixed infections in clinics are common, resulting in significant economic losses in pig industry. Here, a dual priming oligonucleotide (DPO)-based multiplex real-time SYBR Green RT-PCR assay were developed for accurately differentiating PEDV, TGEV, PoRV, and PDCoV in clinical specimens targeting the N gene of TGEV, PEDV, and PDCoV, and the VP7 gene of PoRV. Results showed that the DPO primer allowed a wider annealing temperature range (40–65 °C) and had a higher priming specificity compared to conventional primer, in which more than 3 nucleotides in the 3′- or 5′-segment of DPO primer mismatched with DNA template, PCR amplification efficiency would decrease substantially or extension would not proceed. DPO-based multiplex real-time RT-PCR method had analytical detection limit of 8.63 × 102 copies/μL, 1.92 × 102 copies/μL, 1.74 × 102 copies/μL, and 1.76 × 102 copies/μL for PEDV, TGEV, PoRV, and PDCoV in clinical specimens, respectively. A total of 67...
Source: Molecular and Cellular Probes - Category: Molecular Biology Source Type: research

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In conclusion, the large diversity of agent combinations and disease situations detected in neonatal diarrhea outbreaks of this study stand for a more personalized diagnosis and management advice at a farm level.
Source: Veterinary Microbiology - Category: Veterinary Research Source Type: research
In this study, the diagnostic accuracy of InGenius Gastrointestinal Viral (GV) Elite Panel, a newly developed one-step multiplex real-time RT-PCR assay simultaneously detecting rotavirus, adenovirus and astrovirus, was evaluated retrospectively analyzing an archival collection of 128 stool samples of children hospitalized with acute gastroenteritis. The overall sensitivity and specificity for the GV assay was 100% and 96.2% for rotavirus, 96.9% and 100% for astrovirus, 100% and 100% for adenovirus, respectively. The InGenius GV assay showed a high concordance with the reference methods and was able to detect all tested gen...
Source: Journal of Virological Methods - Category: Virology Authors: Tags: J Virol Methods Source Type: research
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Source: Food and Environmental Virology - Category: Virology Source Type: research
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Source: Food and Environmental Virology - Category: Virology Source Type: research
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Source: Infection, Genetics and Evolution - Category: Genetics & Stem Cells Authors: Tags: Infect Genet Evol Source Type: research
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Source: BMC Veterinary Research - Category: Veterinary Research Authors: Tags: Research article Source Type: research
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Source: Journal of Virological Methods - Category: Virology Authors: Tags: J Virol Methods Source Type: research
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Source: Journal of Clinical Virology - Category: Virology Authors: Source Type: research
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Source: Infection, Genetics and Evolution - Category: Genetics & Stem Cells Authors: Tags: Infect Genet Evol Source Type: research
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Source: Life in the Fast Lane - Category: Emergency Medicine Authors: Tags: Clinical Cases Tropical Medicine cholera diarrhoea john snow ORS rice water diarrhoea watery diarrhoea Source Type: blogs
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