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J+Vet+Diagn+Invest 2019 ; 31 (6): 909-912 Nephropedia Template TP
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One-step triplex reverse-transcription PCR detection of porcine epidemic diarrhea virus, porcine sapelovirus, and porcine sapovirus #MMPMID31650901Jiang C; He H; Zhang C; Zhang X; Han J; Zhang H; Luo Y; Wu Y; Wang Y; Ge B; Xu JJ Vet Diagn Invest 2019[Nov]; 31 (6): 909-912 PMID31650901show ga
Swine diarrhea can be caused by multiple agents, including porcine epidemic diarrhea virus (PEDV), porcine sapelovirus (PSV), and porcine sapovirus (SaV). We designed a one-step triplex reverse-transcription PCR (RT-PCR) detection method including 3 pairs of primers that focused on the S1 gene of PEDV, a conserved gene of PSV, and the VP1 gene of SaV. The optimal concentrations of upstream and downstream primers in the triplex RT-PCR were 0.24 muM for PEDV, 0.15 muM for PSV, and 0.2 muM for SaV, and the optimal annealing temperature was 55.5 degrees C. Triplex RT-PCR assessment of 402 piglet diarrhea samples was compared with conventional individual RT-PCR. Concordance rates in both tests for individual viruses were 100%, 97.6%, and 94.4% for PEDV, PSV, and SaV, respectively. PEDV, PSV, and SaV were detected in 57.2%, 10.4%, and 9.0% of the samples, respectively. The high sensitivity and specificity of this triplex RT-PCR-based detection method for PEDV, PSV, and SaV could allow rapid detection and analysis of mixed infections by these 3 viruses.|Animals[MESH]|Caliciviridae Infections/diagnosis/*veterinary[MESH]|Coronavirus Infections/diagnosis/*veterinary[MESH]|Diarrhea/diagnosis/*veterinary[MESH]|Multiplex Polymerase Chain Reaction/methods/*veterinary[MESH]|Picornaviridae Infections/diagnosis/*veterinary[MESH]|Picornaviridae/isolation & purification[MESH]|Porcine epidemic diarrhea virus/isolation & purification[MESH]|Reverse Transcriptase Polymerase Chain Reaction/methods/*veterinary[MESH]|Sapovirus/isolation & purification[MESH]|Swine[MESH]
  
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