Chu Yue-Feng, Gao Peng-Chen, Zhao Ping, He Yin, Zhang Nian-Zhang, Liu Yong-Sheng, Liu Ji-Xing, Lu Zhong-Xin
Key Laboratory of Grazing Animal Diseases of Ministry of Agriculture, Key Laboratory of Animal Virology of Ministry of Agriculture, State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.
J Vet Med Sci. 2011 Mar;73(3):337-43. doi: 10.1292/jvms.10-0296. Epub 2010 Nov 4.
Outer membrane proteins (OMPs) are the major virulent factors of Haemophilus parasuis. PCR-RFLP targeting the ompA gene was conducted to investigate the possibility of genotyping H. parasuis in this study. Fifteen reference strains and 49 isolates from pig farms in northwest China were genotyped by PCR-RFLP with a pair of specific primers. The results indicated that both the 15 reference strains and 49 isolates could be classified into 8 different genotypes by PCR-RFLP, respectively. Seven genotypes including AA, BB, BA, CA, BC, BD and CD existed simultaneously in the reference strains and isolates, but genotype CB only existed in the isolated strains. Interestingly, genotypes BA, CD and CA were only found in diseased pigs and accounted for 38.8%, 22.4% and 18.4% of the isolates, respectively. On the other hand, strains isolated from apparently healthy pigs were classified into genotypes AA, BB, BC and CB. However, the virulent reference serovar 1 strain has an AA genotype, and the fact that nearly all strains from the healthy pigs belonged to serovars classed as virulent suggests that these genotypes might also include virulent strains; therefore, further validation with more field strains is needed. The capability of the RFLP-PCR method based on the ompA gene for genotyping H. parasuis isolates indicates that this method may be a useful tool for epidemiological study.
外膜蛋白(OMPs)是副猪嗜血杆菌的主要致病因子。本研究采用针对ompA基因的PCR-RFLP技术来探讨副猪嗜血杆菌基因分型的可能性。使用一对特异性引物通过PCR-RFLP对15株参考菌株和来自中国西北部猪场的49株分离株进行基因分型。结果表明,15株参考菌株和49株分离株均可通过PCR-RFLP分别分为8种不同的基因型。参考菌株和分离株中同时存在包括AA、BB、BA、CA、BC、BD和CD在内的7种基因型,但基因型CB仅存在于分离株中。有趣的是,基因型BA、CD和CA仅在患病猪中发现,分别占分离株的38.8%、22.4%和18.4%。另一方面,从外观健康猪分离的菌株被分为基因型AA、BB、BC和CB。然而,强毒参考血清1型菌株具有AA基因型,并且几乎所有来自健康猪的菌株都属于被归类为强毒的血清型,这表明这些基因型可能也包括强毒株;因此,需要用更多的田间菌株进行进一步验证。基于ompA基因的RFLP-PCR方法对副猪嗜血杆菌分离株进行基因分型的能力表明,该方法可能是流行病学研究的有用工具。