摘要
为了解福建地区致猪腹泻病病毒的流行现状和猪流行性腹泻病毒(PEDV)的变异情况,2023—2024年从福州市某疑似感染腹泻病毒的规模化猪场采集发病仔猪样品,经RT-PCR进行PEDV、猪传染性胃肠炎病毒(TGEV)和猪轮状病毒(PoRV)核酸检测,并对PEDV阳性病料进行纤突蛋白(S)基因序列扩增和测序,进行遗传演化分析。结果:PEDV、TGEV和PoRV检出率分别为32.29%(31/96)、30.21%(29/96)和4.17%(4/96),其中PEDV和TGEV混合感染17份(17.71%),TGEV和PoRV混合感染1份(1.04%),PEDV、TGEV和PoRV三重混合感染1份(1.04%)。从10份PEDV阳性猪肠道组织样品中获得10株PEDV的S基因序列,全长3753~4179 bp,编码1250~1392 aa,10株PEDV的S基因核苷酸及氨基酸序列同源性分别为94.9%~99.8%和83.8%~99.7%。S基因系统发育进化树结果表明,6株属于GⅠb亚型,4株属于GⅡb亚型,且4株PEDV的S基因为独立的一小分支。与经典毒株CV777的S基因序列相比,10株PEDV的S基因核苷酸及氨基酸序列同源性分别为93.6%~96.9%和83.9%~95.9%,存在23个保守氨基酸位点突变,在162 aa处缺失1个氨基酸(R),GⅠb亚型的6株PEDV S蛋白存在10个保守氨基酸位点突变;GⅡb亚型的4株PEDV S蛋白存在45个保守氨基酸位点突变,在59~62 aa处有4个氨基酸(QGVN)插入,163 aa处缺失1个氨基酸(D)。综上,福建某规模化猪场PEDV流行率较高,流行毒株基因型呈现多样性,说明流行株持续变异,临床上应予以高度重视。
In order to understand the epidemic status of porcine diarrhea-causing viruses and the variation of porcine epidemic diarrhea virus(PEDV)in Fujian Province,this study collected samples from diseased piglets in a large-scale pig farm suspected of being infected with diarrhea viruses,in Fuzhou City,Fujian Province,from 2023 to 2024.Reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the nucleic acid of PEDV,transmissible gastroenteritis virus of pigs(TGEV)and porcine rotavirus(PoRV).Next,the spike protein(S)gene sequences of the PEDV-positive samples were amplified and sequenced,and genetic evolution and sequence analysis were conducted to identify the genetic variation of the S gene.The results showed that the positive rates of PEDV,TGEV and PoRV were 32.29%(31/96),30.21%(29/96)and 4.41%(4/96),respectively;and there were 17 samples(17.71%)were of PEDV and TGEV mixed infection,1 sample(1.04%)was of TGEV and PoRV mixed infection,1 sample(1.04%)was of PEDV,TGEV and PoRV triple mixed infections.The S gene sequences of ten PEDV strains were obtained fromthe PEDV-positive intestinal tissue samples,with a full length ranging from 3753 bp to 4179 bp,encoding 1250 to 1392 amino acids.The nucleotide and amino acid sequence homologies among the S genes of the 10 PEDV strains were 94.9%-99.8%and 83.8%-99.7%,respectively.The results of the phylogenetic tree of the S gene indicated that 6 strains belonged to the GⅠb subtype,and 4 strains belonged to the GⅡb subtype.Additionally,the S gene of 4 PEDV strains formed a separate small branch.Compared with the S gene sequence of CV777,the nucleotide and amino acid sequence homologies of S genes of the 10 PEDV strains were 93.6%-96.9%and 83.9%-95.9%,respectively.And a total of 23 conserved amino acid mutations were identified,including a deletion of 1 amino acid(R)at position 162 aa.Among the GⅠb subtype strains,10 amino acid mutations were observed in the S protein,while the GⅡb subtype strains exhibited 45 amino acid mutations,including an insertion of 4 amino acids(QGVN)at positions 59-62 aa and a deletion of 1 amino acid(D)at position 163 aa.These results indicated that the prevalence of PEDV is relatively high in the large-scale pig farm in Fujian Province,and the genotypes of the prevalent strains are diverse,suggesting that the prevalent strains are continuously mutating.
作者
郭桂兰
王凯莉
王鑫鑫
郭桂杰
GUO Guilan;WANG Kaili;WANG Xinxin;GUO Guijie(College of Animal Sciences,Fujian Agriculture and Forestry University,Fuzhou 350002,China)
出处
《畜牧与兽医》
北大核心
2025年第7期131-138,共8页
Animal Husbandry & Veterinary Medicine
基金
福建省自然科学基金面上项目(2024J01388)。
关键词
猪流行性腹泻病毒
猪轮状病毒
猪传染性胃肠炎病毒
鉴定
序列分析
福州
porcine epidemic diarrhea virus
porcine rotavirus
transmissible gastroenteritis virus of swine
identification
sequence analysis
Fuzhou
