Wang Siying, Hu Xia, Xiong Tao, Cao Lijing, Zhang Xingcui, Song Zhenhui
College of Veterinary Medicine, Southwest University, Rongchang, Chongqing, 402460, China.
Animal Epidemic Prevention and Quarantine Center of Cuiping District, Yibin, 644000, China.
BMC Vet Res. 2024 Dec 19;20(1):565. doi: 10.1186/s12917-024-04390-4.
In recent years, the prevalence and incidence of porcine epidemic diarrhea virus (PEDV) infection have been on the rise. The occurrence of multiviral infections and recombination mutations has led to accelerated viral evolution and reduced vaccine efficacy. In the present study, a PEDV strain was isolated from a pig farm (Chongqing Province, China) with an outbreak of porcine diarrhea, and its S gene was found to be recombinant.
The optimal trypsin concentration for blind passage of PEDV in Vero cells was determined to be 7.5 µg/mL. Following two blind passages of the virus in Vero cells, the virus was unable to adapt to the cells. Therefore, PEDV was blindly passaged in IPEC-J2 cells using the optimal concentration of trypsin (5 µg/mL). Next, a series of characterization experiments were performed. Recombination analyses of the isolates using software revealed that the S gene of strain CHCQ-2023 was derived from the primary parent strain PEDV-1 C and secondary parent strain SQ2014, with recombination occurring at a 3152 bp breakpoint. Furthermore, a specific B-cell antigenic epitope was predicted on the S2 subunit of the S protein.
A PEDV strain was isolated and characterized, and its S gene was characterized. The findings provide a bioinformatic basis for the study of PEDV strain variation due to genetic recombination.
近年来,猪流行性腹泻病毒(PEDV)感染的流行率和发病率一直在上升。多种病毒感染和重组突变的发生导致病毒进化加速和疫苗效力降低。在本研究中,从中国重庆市一个发生猪腹泻疫情的猪场分离出一株PEDV毒株,发现其S基因是重组的。
确定PEDV在Vero细胞中盲目传代的最佳胰蛋白酶浓度为7.5 μg/mL。该病毒在Vero细胞中盲目传代两次后,无法适应这些细胞。因此,使用最佳浓度的胰蛋白酶(5 μg/mL)在IPEC-J2细胞中对PEDV进行盲目传代。接下来,进行了一系列鉴定实验。使用软件对分离株进行重组分析表明,CHCQ-2023毒株的S基因来源于初级亲本毒株PEDV-1 C和次级亲本毒株SQ2014,重组发生在3152 bp的断点处。此外,在S蛋白的S2亚基上预测到一个特异性B细胞抗原表位。
分离并鉴定了一株PEDV毒株,并对其S基因进行了表征。这些发现为研究PEDV毒株因基因重组而发生的变异提供了生物信息学基础。