The Complex Co-infections of Multiple Porcine Diarrhea Viruses in Local Area Based on the Luminex xTAG Multiplex Detection Method

Abstract

The large-scale outbreaks of severe diarrhea caused by viruses have occurred in pigs since 2010, resulting in great damage to the pig industry. However, multiple infections have contributed to the outbreak of the disease and also resulted in great difficulties in diagnosis and control of the disease. Thus, a Luminex xTAG multiplex detection method, which was more sensitive and specific than general multiplex PCR method, was developed for the detection of 11 viral diarrhea pathogens, including PKoV, PAstV, PEDV, PSaV, PSV, PTV, PDCoV, TGEV, BVDV, PoRV, and PToV. To investigate the prevalence of diarrhea-associated viruses responsible for the outbreaks, a total of 753 porcine stool specimens collected from 9 pig farms in Shanghai during 2015–2018 were tested and the pathogen spectrums and co-infections were analyzed. As a result, PKoV, PAstV and PEDV were most commonly detected viruses in diarrheal pigs with the rate of 38.65% (291/753), 20.32% (153/753), and 15.54% (117/753), respectively. Furthermore, multiple infections were commonly seen, with positive rate of 28.42%. Infection pattern of the viral diarrhea pathogens in a specific farm was changing, and different farms had the various diarrhea infection patterns. A longitudinal investigation showed that PEDV was the key pathogen which was closely related to the death of diarrhea piglets. Other pathogens might play synergistic roles in the pathogenesis of diarrhea disease. Furthermore, the surveillance confirmed that variant enteropathogenic viruses were leading etiologic agents of porcine diarrhea, either mono-infection or co-infections of PKoV were common in pigs in Shanghai, but PEDV was still the key pathogen and multiple pathogens synergistically complicated the infection status, suggesting that controlling porcine diarrhea might be more complex than previously thought. The study provides a better understanding of diarrhea viruses in piglets, which will aid in better preventing and controlling epidemics of viral porcine diarrhea.

Occurrence of Rotavirus A Genotypes and Other Enteric Pathogens in Diarrheic Suckling Piglets from Spanish Swine Farms

“…It is worth mentioning that a majority of cases (>80%) corresponded to mixed infections with different combinations of the target agents, a finding which has also been highlighted in previous studies in Spanish herds [5,8]. Diarrhea in a herd may be due to a single agent, but the finding of multiple concurrent agents is being reported increasingly in clinics, frequently with complex interactions that can result in synergistic or additive effects leading to more severe and prolonged diarrhea [27,28]. Enterotoxigenic E. coli (ETEC) strains were not included in the diagnostic panel in the current study due to earlier studies indicating that their prevalence is very low in neonatal piglet diarrhea in Spanish herds.…”

Advances in the differential molecular diagnosis of vesicular disease pathogens in swine

“…The final RPA product is applied to the LFA strip, and the results are visualized after the appearance of matching lanes (Bhat et al, 2022(Bhat et al, ). 10.3389/fmicb.2022 Frontiers in Microbiology frontiersin.org simultaneous detection of 11 viral diarrhea pathogens (Shi et al, 2021). A total of 753 porcine stool specimens from five districts in Shanghai were detected using this assay and the surveillance confirmed that PEDV is still the main pathogen of porcine diarrhea (Shi et al, 2021).…”

“…Wu et al developed a PCR-based fluorescent Luminex assay for human papillomavirus (HPV) genotyping, and the assay may be used to provide critical clinical information for the early detection of HPV ( Oh et al, 2007 ). A Luminex xTAG multiplex detection method with high sensitivity and specificity, was developed for the simultaneous detection of 11 porcine viral diarrhea pathogens ( Shi et al, 2021 ). A total of 753 porcine stool specimens from five districts in Shanghai were detected using this assay and the surveillance confirmed that PEDV is still the main pathogen of porcine diarrhea ( Shi et al, 2021 ).…”

Establishment of a Multiplex RT-PCR Method for the Detection of Five Known Genotypes of Porcine Astroviruses

“…However, emerging cases of PAstV associated enteritis or polioencephalomyelitis were reported and attracted public attention in recent years ( 8 , 9 , 34 , 35 ). As an enteric virus, PAstVs were more frequently detected in the pig herds, and the prevalent rates of PAstV were usually reported much higher than those of other diarrheal viruses such as PEDV, TGEV, and porcine deltacoronavirus (PDCoV) ( 30 , 36 ). Besides, the high genetic variability and possible recombination events of PAstVs further remind people to develop comprehensive methods for astrovirus diagnosis and epidemiological investigation ( 23 , 37 ).…”