Development of a multiplex qRT-PCR assay for the detection of porcine epidemic diarrhea virus, porcine transmissible gastroenteritis virus and porcine Deltacoronavirus

Abstract

Currently, porcine coronaviruses are prevalent in pigs, and due to the outbreak of COVID-19, porcine coronaviruses have become a research hotspot. porcine epidemic diarrhea virus (PEDV), Transmissible Gastroenteritis Virus (TGEV), and Porcine Deltacoronavirus (PDCoV) mentioned in this study mainly cause diarrhea in pigs. These viruses cause significant economic losses and pose a potential public health threat. In this study, specific primers and probes were designed according to the M gene of PEDV, the S gene of TGEV, and the M gene of PDCoV, respectively, and TaqMan probe-based multiplex real-time quantitative reverse transcription-polymerase chain reaction (qRT-PCR) was developed for the simultaneous detection of PEDV, TGEV, and PDCoV. This method has high sensitivity and specificity, and the detection limit of each virus can reach 2.95 × 100 copies/μl. An assay of 160 clinical samples from pigs with diarrhea showed that the positive rates of PEDV, TGEV, and PDCoV were 38.13, 1.88, and 5.00%; the coinfection rates of PEDV+TGEV, PEDV+PDCoV, TGEV+PDCoV, PEDV+TGEV+PDCoV were 1.25, 1.25, 0, 0.63%, respectively. The positive coincidence rates of the multiplex qRT-PCR and single-reaction qRT-PCR were 100%. This method is of great significance for clinical monitoring of the porcine enteric diarrhea virus and helps reduce the loss of the breeding industry and control the spread of the disease.

Development of a multiplex reverse transcription-quantitative PCR (qPCR) method for detecting common causative agents of swine viral diarrhea in China

“…The lowest detectable copy numbers of the method assessed using synthesized plasmids were lower than 10 copies/µL (3 copies/µL for PEDV, 4 copies/µL for TGEV, 8 copies/µL for RVA, and 8 copies/µL for PDCoV). These detection limits were similar to those (2.95 × 10 0 copies/µL) in a documented multiplex qPCR which was developed to detect PEDV, TGEV, and PDCoV simultaneously [ 26 ]. These detection limits were much lower than those reported in another multiplex qPCR which was developed for detecting different types of PEDV (20 copies/µL for GI and 100 copies/µL for GII, 50 copies/µL for both RVA and RVC) [ 27 ].…”

A novel host restriction factor MRPS6 mediates the inhibition of PDCoV infection in HIEC-6 cells

“…The primary sites of infection are the distal jejunum and ileum, leading to symptoms such as watery diarrhea, vomiting, dehydration, and weight loss ( 19 , 20 ). Their clinical manifestations are similar to those caused by porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), and co-infections ( 21 ). This virus poses a significant threat to the livestock industry, particularly due to its high piglet mortality rates.…”

“…This virus poses a significant threat to the livestock industry, particularly due to its high piglet mortality rates. PDCoV exhibits a broad spectrum of infectivity across multiple species, including pigs, poultry, mice, leopard cats, pigeons, and even humans, indicating a potential risk to public health (21)(22)(23)(24)(25)(26).…”

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“…The primary sites of infection are the distal jejunum and ileum, leading to symptoms such as watery diarrhea, vomiting, dehydration, and weight loss (19,20). Their clinical manifestations are similar to those caused by porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), and co-infections (21). This virus poses a significant threat to the livestock industry, particularly due to its high piglet mortality rates.…”

“…This virus poses a significant threat to the livestock industry, particularly due to its high piglet mortality rates. PDCoV exhibits a broad spectrum of infectivity across multiple species, including pigs, poultry, mice, leopard cats, pigeons, and even humans, indicating a potential risk to public health (21)(22)(23)(24)(25)(26).…”