贵州省2017年~2023年PEDV和PoRVA流行病学调查及PoRVA VP7、VP4基因的遗传演化分析  

Epidemiological investigation on the prevalence of PEDV and PoRVA in Guizhou during 2017-2023 and genetic evolution analysis of VP7 and VP4 genes of PoRVA strains.

作　　者：田红利 柳佳佳 梁海英[1] 曾智勇[1] 汤德元[1] 王彬[1] 边孟婷 黄书 潘向英 TIAN Hong-li;LIU Jia-jia;LIANG Hai-ying;ZENG Zhi-yong;TANG De-yuan;WANG Bin;BIAN Meng-ting;HUANG Shu;PAN Xiang-ying(College ofAnimal Science,Guizhou University,Guiyang 550025,China)

机构地区：[1]贵州大学动物科学学院,贵州贵阳550025

出　　处：《中国预防兽医学报》2024年第8期791-798,共8页Chinese Journal of Preventive Veterinary Medicine

基　　金：“生猪常见病毒性疫病快速检测及综合防控技术研究与示范”黔科合支撑[2021]一般162项目资助。

摘　　要：为了解贵州省猪流行性腹泻病毒(PEDV)和猪A群轮状病毒(PoRVA)的流行情况,本研究对2017年~2023年来自贵州省9个地区139个猪场的165份仔猪粪便样品进行Taq Man RT-qPCR检测,并对PoRVA阳性样品VP7、VP4基因进行RT-PCR扩增、测序,遗传进化分析及其编码氨基酸序列的变异分析。结果显示,PEDV、PoRVA和PEDV+PoRVA的感染率分别为67.88%(112/165)、38.79%(64/165)和21.21%(35/165),且在不同年份、不同地区和不同季节均检出三者的感染。PCR结果显示,本实验扩增出27株PoRVA VP7基因和22株VP4基因;遗传进化分析结果显示共鉴定出6种G型和3种P型,其中G9(29.63%)和P[13](68.18%)为优势基因型,G3、G4、G5、G11、G26、P[6]和P[23]分别占7.41%、22.22%、22.22%、3.70%、14.81%、13.64%、18.18%。21份PoRVA样品鉴定出10种G/P组合基因型,G9P[13](19.05%)为优势组合基因型,G4P[13]、G5P[13]、G3P[13]、G4P[6]、G5P[23]、G9P[23]、G11P[13]、G26P[6]、G26P[13]分别占14.29%、14.29%、9.52%、9.52%、9.52%、9.52%、4.76%、4.76%及4.76%。同源性分析结果显示,27株VP7基因和22株VP4基因序列与NX疫苗株相应基因序列之间的同源性分别为75.5%~94.0%和68.5%~75.6%,氨基酸序列的同源性分别为78.3%~96.6%和72.0%~82.5%,部分PoRVA上述基因序列与国外、人源RVA的同源性最高。氨基酸位点变异分析结果显示,与NX疫苗株相比,所测PoRVA VP7和VP4氨基酸序列均存在特征性位点的变异,其中G型PoRVA VP7无氨基酸位点的缺失和插入,G9型PoRVA VP7存在3处(I^(208)T、K^(212)A/V/T、V^(271)I)变异,G3、G4、G5、G11和G26型PoRVA VP7存在17处(S^(28)R/K、A^(29)I/T/M/V、L^(40)I/V、V^(44)L、S^(71)T、Q^(73)G/N/S/E、S^(90)A/Q/K/R、G^(94)N/S/A/K、N^(100)D/E、T^(122)A/S、T^(147)A/G/N/D、Q^(189)S/T、I^(208)L/Q/T、K^(212)T/I/P/N、A^(213)N/T/D、E^(267)D/N、V^(271)I)变异;3株P[6]型PoRVA VP4在aa136缺失T,13株P[13]型PoRVA VP4在aa188~aa189插入E/D~Y/F,P[6]、P[13]和P[23]型PoRVA VP4存在26处(I^(92)V、Q^(9To understand the prevalence of porcine epidemic diarrhea virus(PEDV)and porcine group A rotavirus(PoRVA)in Guizhou Province,165 piglet diarrhea samples from 139 pig farms in 9 regions of Guizhou Province from 2017 to 2023 were detected by TaqMan RT-qPCR,and RT-PCR amplification,sequencing,and genetic evolution analysis of VP7 and VP4 genes of PoRVA-positive samples were performed.The results showed that the infection rates of PEDV,PoRVA and PEDV+PoRVA were 67.88%(112/165),38.79%(64/165)and 21.21%(35/165),respectively,and the infections of the above three were detected in different years,regions and seasons.Genetic evolution analysis showed that 27 strains of PoRVA VP7 gene and 22 strains of VP4 gene were amplified and identified as 6 G-genotype and 3 P-genotype,among which G9(29.63%)and P[13](68.18%)were the dominant genotypes,and G3,G4,G5,G11,G26,P[6]and P[13]accounted for 7.41%,22.22%,22.22%,3.70%,14.81%,13.64%and 18.18%,respectively.Ten genotypes of G/P combinations were successfully identified in 21 PoRVA samples,among which G9P[13](19.05%)was the dominant combination genotype and G4P[13],G5P[13],G3P[13],G4P[6],G5P[23],G9P[23],G11P[13],G26P[6],G26P[13]accounting for 14.29%,14.29%,9.52%,9.52%,9.52%,9.52%,4.76%,4.76%,4.76%,respectively.The results of homology analysis showed that the homology between 27 strains of the VP7 gene and 22 strains of the VP4 gene with the NX vaccine strain,were 75.5%-94.0%and 68.5%-75.6%,and the homology of amino acids were 78.3%-96.6%and 72.0%-82.5%,respectively.Some PoRVA sequences had the highest homology with foreign and human-derived group A rotaviruses.The results of amino acid site variation analysis showed that compared to the NX vaccine strain,the amino acid sequences of PoRVA VP7 and VP4 all showed characteristic amino acid site mutations.There were no deletions or insertions of amino acid sites in G-genotype PoRVA VP7,3 mutations(I^(208)T,K^(212)A/V/T,V^(271)I)in G9 genotype PoRVA VP7,and 17 mutations(S^(28)R/K,A^(29)I/T/M/V,L^(40)I/V,V^(44)L,S^(71)T,Q^(73)G/N/S/E,S^(90)A

关 键 词：猪流行性腹泻病毒 猪A群轮状病毒 分子流行病学 VP7和VP4基因 遗传演化分析 

分 类 号：S852.65[农业科学—基础兽医学]