A TaqMan Probe-Based Multiplex Real-Time PCR for Simultaneous Detection of Porcine Epidemic Diarrhea Virus Subtypes G1 and G2, and Porcine Rotavirus Groups A and C

Abstract

Porcine viral diarrhea diseases affect the swine industry, resulting in significant economic losses. Porcine epidemic diarrhea virus (PEDV) genotypes G1 and G2, and groups A and C of the porcine rotavirus, are major etiological agents of severe gastroenteritis and profuse diarrhea, particularly among piglets, with mortality rates of up to 100%. Based on the high prevalence rate and frequent co-infection of PEDV, RVA, and RVC, close monitoring is necessary to avoid greater economic losses. We have developed a multiplex TaqMan probe-based real-time PCR for the rapid simultaneous detection and differentiation of PEDV subtypes G1 and G2, RVA, and RVC. This test is highly sensitive, as the detection limits were 20 and 100 copies/μL for the G1 and G2 subtypes of PEDV, respectively, and 50 copies/μL for RVA and RVC, respectively. Eighty-eight swine clinical samples were used to evaluate this new test. The results were 100% in concordance with the standard methods. Since reassortment between porcine and human rotaviruses has been reported, this multiplex test not only provides a basis for the management of swine diarrheal viruses, but also has the potential to impact public health as well.

Recent Molecular Characterization of Porcine Rotaviruses Detected in China and Their Phylogenetic Relationships with Human Rotaviruses

“…In 2022, a study conducted in Heilongjiang province reported an RVA positive rate of 4.3% (12/280) [ 25 ]. Additionally, a study conducted in Jiangsu province in 2022 reported an RVA positive rate of 12.5% (11/88) [ 26 ]. For a comprehensive understanding of the prevalence of PoRVA in China, a large number of porcine diarrheal samples were collected from the major provinces in pig production.…”

Genome Analysis of the G6P6 Genotype of Porcine Group C Rotavirus in China

“…In recent years, the number of porcine RVC genomes has increased significantly, indicating that it has higher genetic diversity than other host species [ 22 ]. The positive rate for PoRVC was detected using a TaqMan Probe-Based Multiplex Real-Time PCR, indicating that the prevalence rate of PoRVC had been previously underestimated [ 23 ]. To facilitate the classification of rotavirus strains, a Rotavirus Classification Working Group (RCWG) was formed to set diagnostic guidelines and propose a classification system [ 24 ].…”

Equine Rotavirus A under the One Health Lens: Potential Impacts on Public Health

“…While the window for the detection of viral shedding using antigen-based detection methods is relatively short (up to one week after onset of diarrhea), viral nucleic acids can be detected for longer periods (e.g., via RT-PCR) [ 248 ]. Currently, multiplex real-time quantitative RT-PCR assays for the simultaneous detection of different agents associated with acute gastroenteritis including RVA have been widely implemented [ 249 , 250 , 251 ].…”