贵州省地方猪肺炎支原体鉴定及其主要黏附因子基因序列分析  

Identification of endemic Mycoplasma hyopneumoniae in Guizhou Province and gene sequence analysis of main adhesion factors

作　　者：张静 赵春萍 张雄 王婧 齐婧 周思旋 杜春林 罗意 史开志 ZHANG Jing;ZHAO Chunping;ZHANG Xiong;WANG Jing;QI Jing;ZHOU Sixuan;DU Chunlin;LUO Yi;SHI Kaizhi(Institute of Animal Husbandry and Veterinary Medicine,Guizhou Academy of Agricultural Sciences,Guiyang 550002,China;Guizhou Firstv Biological Technology Co.,Ltd.,Guiyang 550002,China)

机构地区：[1]贵州省农业科学院畜牧兽医研究所,贵阳550002 [2]贵州福斯特生物科技有限公司,贵阳550002

出　　处：《黑龙江畜牧兽医》2022年第5期73-81,共9页Heilongjiang Animal Science And veterinary Medicine

基　　金：贵州省科技厅成果转化项目(黔科合成果[2020]1Y027号);中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室开放基金项目(SKLVEB2019KFKT015);贵州省农业科学院畜牧兽医研究所创新基金项目(黔牧医所创新基金[2018]03-02号);贵州省农业科学院青年基金项目(黔农科院青年基金[2018]92号)。

摘　　要：为了了解贵州地方猪肺炎支原体菌株遗传变异情况,试验对3份贵州省贵阳市某地方猪养殖场疑似感染猪肺炎支原体的病料进行离体培养及PCR鉴定;同时对分离菌株及6株疫苗菌株的主要黏附因子P46基因、P97基因R1区和P146基因高变区进行PCR扩增及测序,并与参考菌株比对分析,挖掘分离菌株与疫苗菌株、参考菌株之间的差异性。结果表明:从疑似病料中培养并鉴定获得1株猪肺炎支原体,命名为GZ株。GZ株P46基因核苷酸序列与疫苗菌株、参考菌株的同源性最高,均在98.3%以上。P146蛋白高变区氨基酸差异主要发生在PQ和PS重复区,GZ株PQ重复区中只缺失2个氨基酸,而PS重复区中缺失7个氨基酸,与疫苗菌株168L株、RM48株和Z株缺失情况一致。GZ株P97基因R1区核苷酸序列与疫苗菌株、参考菌株之间的同源性低,为94.5%~97.4%;各菌株间P97蛋白R1区氨基酸序列中,AAKPV/E重复基元存在不同程度缺失,GZ株在该区域仅存在5个重复基元,且第1个重复序列中的谷氨酸(E)突变为谷氨酰胺(Q)。说明P97蛋白R1区和P146蛋白高变区氨基酸序列的改变导致抗原表位和黏附能力发生变化,对P97基因R1区和P146基因高变区遗传变异情况的调查将有助于筛选出最佳商品化疫苗和研制地方性适用疫苗,有效阻止肺炎支原体在猪群中的传播。In order to understand the genetic variation of endemic Mycoplasma hyopneumoniae strains in Guizhou Province, the experiment conducted in vitro culture and PCR identification of 3 diseased samples suspected to be infected with Mycoplasma hyopneumoniae in a local pig breeding farm in Guiyang City, Guizhou Province;at the same time, PCR amplification and sequencing were carried out based on the main adhesion factors of P46 gene, P97 gene R1 region and P146 gene hypervariable region of the isolated strains and 6 commercial vaccines;and comparison analysis was performed with the reference strains to discover the differences between the isolated strains and the vaccine strains or the reference strains. The results showed that a strain of Mycoplasma hyopneumoniae was obtained from the suspected disease material and named as GZ strain. The nucleotide sequence of the P46 gene of the GZ strain had the highest homology with the vaccine strain and the reference strain, both of which were above 98.3%. The amino acid difference in the P146 gene hypervariable region mainly occurred in the PQ and PS repeats;only 2 amino acids were missing in the PQ repeat region of the GZ strain, while 7 amino acids were missing in the PS repeat region, which was consistent with the deletion of vaccine strains 168 L, RM48 and Z. The results suggested that the changes in the amino acid sequence of the R1 region of the P97 gene and the hypervariable region of the P146 gene led to variation in the epitope and adhesion ability. The homology between the nucleotide sequence of the P97 gene R1 region of the GZ strain and the vaccine strain and the reference strain was low, ranging from 94.5% to 97.4%. In the amino acid sequence of the P97 gene R1 region among the strains, the AAKPV/E repeat motif had varying degrees of deletion;the GZ strain only had 5 repeat motifs in this region, and the glutamate(E) of the first repeat motif was mutated to glutamine(Q). The investigation of the genetic variation in the R1 region of the P97 gene and the hypervariabl

关 键 词：猪肺炎支原体 GZ株 疫苗菌株 P46基因 P146基因 P97基因 

分 类 号：S852.[农业科学—基础兽医学]