副猪格拉瑟菌的荚膜分型方法及原理研究进展  

Advances in Capsular Typing Method and Principle of Glaesserella parasuis

作　　者：朱亚新 关丽君 张俊峰[1] 薛云[1] 赵战勤[1,2] ZHU Yaxin;GUAN Lijun;ZHANG Junfeng;XUE Yun;ZHAO Zhanqin(Key Lab of Animal Bacterial Infectious Disease Prevention and Control Technology,College of Animal Science and Technology,Henan University of Science and Technology,Luoyang 471003,China;Key-Disciplines Lab of safety of environment and Animal Product,College of Animal Science and Technology,Henan University of Science and Technology,Luoyang 471003,China)

机构地区：[1]河南科技大学动物科技学院/洛阳市动物细菌性传染病防控技术重点实验室,洛阳471003 [2]河南科技大学动物科技学院/河南省高等学校环境与畜产品安全重点学科开放实验室,洛阳471003

出　　处：《畜牧兽医学报》2024年第12期5412-5422,共11页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金项目(31672530,U1704117)。

摘　　要：副猪格拉瑟菌(GPS)是猪的重要病原菌之一,自1910年被确认为猪格拉瑟病的病原以来,血清型鉴定是GPS研究领域长期以来的“卡脖子”环节。1992年,基于GPS热稳定抗原的凝胶免疫扩散试验(GID)分型方法首次将GPS分为1-15型,但有约25%的菌株不可分型,当时对于其分型抗原的成分也不明确。2003年,基于GPS的“盐水提取物”抗原建立了GPS的间接血凝试验(IHA)分型方法,其敏感性和分型率均高于GID,但仍有约15%的菌株不可分型。IHA和GID方法的分型结果基本一致,因此能够确定两者的分型抗原是基于相同的GPS表面多糖成分,但仍不能确定是荚膜多糖,或是LPS,甚或是其他多糖物质。2013年,1-15型GPS荚膜多糖的合成基因簇得到成功解析,证实其血清型抗原形成的本质是荚膜多糖。2015年,Howell等基于GPS荚膜的特异性靶基因设计引物,建立了GPS的PCR分型方法(H-PCR),但不能对血清5型和12型进行鉴别;2017年,Jia等基于GPS荚膜的特异性靶基因也建立了一套PCR分型方法(J-PCR),能对血清5型和12型进行了鉴别。两种PCR分型方法配合使用可对几乎所有GPS菌株分型,并从分子生物学基础上验证了其与GID和IHA方法的一致性,最终确认三种分型方法的抗原基础均为GPS荚膜。PCR分型方法具有操作简单、速度快、成本低等优点,成功解决了GID和IHA血清分型方法中的诸多问题,得到了广泛应用。本文系统回顾了GPS GID、IHA和PCR分型方法的建立与应用历史,及其相同抗原基础的发现历程,以期为GPS三种分型方法的免疫学物质基础及原理提供系统的理论知识,为副猪格拉瑟菌病防控技术的研发提供新思路。Glaesserella parasuis(GPS)is one of the important pathogenic bacteria in pigs,and serotype identification has been a long-standing"bottleneck"in the field of GPS research since it was identified as the causative agent of Glasser's disease in pigs in 1910.In 1992,the gel immunodiffusion(GID)typing method based on GPS heat-stable antigens classified GPS as 1-15 for the first time,but about 25%of the strains were not typed,and the composition of the typed antigen was not clear at that time.In 2003,an indirect hemagglutination assay(IHA)typing method for GPS-based"saline extract"antigen was established,and its sensitivity and typing rate were higher than those of GID,but about 15%of the strains were still untypeable.The typing results of IHA and GID methods are generally identical,so it is possible to determine whether the typed antigens are based on the same GPS surface polysaccharide composition,but it is still not certain whether it is a capsular polysaccharide,or LPS,or even other polysaccharide species.In 2013,the synthetic gene cluster of GPS serotype 1-15 capsular polysaccharide was successfully analyzed,which confirmed that the essence of serotype antigen formation was capsular polysaccharide.In 2015,Howell et al.designed primers based on the specific target genes of GPS capsule,and established a PCR typing method for GPS(H-PCR),but it could not distinguish serotype 5 and 12.In 2017,Jia et al.also established a set of PCR typing methods(J-PCR)for specific target genes based on GPS capsule,which could identify serotypes 5 and 12.The combination of the two PCR typing methods was able to type almost all GPS strains,and their consistency with the GID and IHA methods was verified from molecular biology,and the antigen basis of the three typing methods was finally confirmed to be GPS capsule.The PCR typing method has the advantages of simple operation,fast speed and low cost,and has successfully solved many problems in GID and IHA serotyping methods,and has been widely used.In this paper,we systematically reviewed

关 键 词：副猪格拉瑟菌 荚膜 血清型 凝胶免疫扩散试验(GID) 间接血凝试验(IHA) PCR 

分 类 号：S852.61[农业科学—基础兽医学]