PCR结合酶切-序列比对法鉴定B群脑膜炎奈瑟菌菌株.DOCVIP

PCR结合酶切-序列比对法鉴定B群脑膜炎奈瑟菌菌杨英英，罗树权，于旭博，乔瑞洁，冯宜扬，刘方蕾，吴兵，谭小梅，赵志强，谢贵林 兰州生物制品研究所有限责任公司第一研究室，甘肃省疫苗工程技术研究中心，甘肃，兰州730046 摘要：目的 B群脑膜炎奈瑟菌菌。方法 用玻片凝集法15株B群脑膜炎奈瑟菌菌株PCR结合酶切法唾液酸转移酶siaD基因，用BLAST软件将PCR产物测序结果与GeneBank 中原始siaD序列比对。 结果 15株菌株玻片凝集结果均为阳性15株菌株的PCR产物均为460 bp；TaqⅠ酶切后，13株菌株的酶切产物仍为460 bp，其PCR产物测序比对结果与B群脑膜炎奈瑟菌原始siaD序列同源性均达到99%；其余2株酶切产物与C群脑膜炎奈瑟菌siaD基因序列同源性分别为98%和99%。 结论 15株菌株PCR结合酶切，13株为B群脑膜炎奈瑟菌菌株，2株为C群脑膜炎奈瑟菌菌株B群脑膜炎奈瑟菌菌。 关键词：B群脑膜炎奈瑟菌；菌种定；唾液酸转移酶基因；聚合酶链反应 中图分类号:378.1+5文献标志码:A 文章编号: 1005-5673(2016) DOI: dentification of Neisseria meningitidis serogroup B strain using PCR amplification, enzyme digestion and sequence alignment YANG Ying-ying*, LUO Shu-quan, YU Xu-bo , QIAO Rui-jie,FENG Yi-yang, LIU Fang-lei, WU Bing, TAN Xiao-mei, ZHAO Zhi-qiang, XIE Gui-lin *The First Research Department in Lanzhou Institute of Biological Products Co., Ltd., Center for Gansu Provincial Vaccine Engineering Research, Lanzhou 730046, Gansu Province, China Abstract：Objective To identify Neisseria meningitidis serogroup B strain by PCR amplification, enzyme digestion and sequence alignment. Methods A total of 15 strains obtained from differert resources were preliminary tested by serological silde agglutination method, and then identified by PCR amplification, enzyme digestion and sequence alignment, namely, the sialyltransferase (SiaD) gene was amplified by PCR. Meanwhile, the 15 PCR amplification products were enzyme -digested and compared with the original sequence of siaD gene in GeneBank by using the BLAST software. Results The silde agglutination test showed all positive for 15 strains. The PCR amplification products all were 460bp for 15 strains. The products of 13 strains remained 460bp after digested by Taq1, and their’s sequence homology were both 99% in comparison with the original sequence of siaD gene in Neisseria meningitidis serogroup B in GeneBank. The enzyme digested products were about 200bp for the other 2 strains, and their’s sequence homology were 98% and 99%, respectively, c