Establishment and Preliminary Application of Duplex RT-PCR for Detection of Emerging Porcine deltacoronavirus and Porcine transmissible gastroenteritis virus
Abstract Porcine deltacoronavirus (PDCoV) is a newly discovered coronavirus. It mainly infects piglets and causes severe enteritis accompanied by diarrhea and vomiting and other symptoms. The symptoms are symptomatically indistinguishable from those that caused by Porcine transmissible gastroenteritis virus (TGEV). In clinic the co-infection of PDCoV and TGEV is common, which causes huge economic losses to the swine industry. In order to control and prevent effectively these diseases, development of rapid detection methods of PDCoV and TGEV is the primary task. Thus, according to the analysis of PDCoV and TGEV N gene sequences in GenBank, two pairs of primers targeting the N gene of PDCoV and TGEV were designed respectively. By optimizing the PCR reaction conditions, a duplex RT-PCR detection method was established for the simultaneous detection of PDCoV and TGEV, and the specificity, sensitivity and repeatability of the method were studied. The results showed that the duplex RT-PCR had high sensitivity and specificity with a limited detection of 3.14×102 copies/μL for PDCoV and 3.68×103 copies/μL for TGEV, respectively, and no specific amplifications from other virus, such as Porcine epidemic diarrhea virus (PEDV), Porcine bocavirus (PBoV), Porcine respiratory and reproductive syndrome virus (PRRSV) or Porcine pseudorabies virus (PRV). 252 clinical samples tested by the duplex RT-PCR showed that the positive rates of PDCoV and TGEV were 12.30% and 4.76%, respectively, co-infection rate of PDCoV and TGEV was 0.79%. The date showed that the duplex RT-PCR method established in this study could be used for the clinical detection of PDCoV and TGEV, which provides the theoretical basis information and technical support for further applied to clinical differential diagnosis and epidemiological investigation of TGEV and PDCoV.
LIU Ling-Ling,CAO Bei-Bei,ZHANG Li-Wei, et al. Establishment and Preliminary Application of Duplex RT-PCR for Detection of Emerging Porcine deltacoronavirus and Porcine transmissible gastroenteritis virus[J]. , 2016, 24(12): 1955-1963.
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