Jelfs J, Munro R, Wedege E, Caugant D A
WHO Collaborating Centre for Reference and Research on Meningococci, Department of Bacteriology, National Institute of Public Health, N-0403 Oslo, Norway.
Clin Diagn Lab Immunol. 2000 May;7(3):390-5. doi: 10.1128/CDLI.7.3.390-395.2000.
The ET-15 clone within the electrophoretic type (ET)-37 complex of Neisseria meningitidis was first detected in Canada in 1986 and has since been associated with outbreaks of meningococcal disease in many parts of the world. While the majority of the strains of the ET-37 complex are serosubtype P1.5,2, serosubtype determination of ET-15 strains may often be incomplete, with either only one or none of the two variable regions (VRs) of the serosubtype PorA outer membrane protein reacting with monoclonal antibodies. DNA sequence analysis of the porA gene from ET-15 strains with one or both unidentified serosubtype determinants was undertaken to identify the genetic basis of the lack of reaction with the monoclonal antibodies. Fourteen different porA alleles were identified among 38 ET-15 strains from various geographic origins. The sequences corresponding to subtypes P1.5a,10d, P1.5,2, P1.5,10d, P1.5a,10k, and P1.5a,10a were identified in 18, 11, 2, 2, and 1 isolate, respectively. Of the remaining four strains, which all were nonserosubtypeable, two had a stop codon within the VR1 and the VR2, respectively, while in the other two the porA gene was interrupted by the insertion element, IS1301. Of the strains with P1.5,2 sequence, one had a stop codon between the VR1 and VR2, one had a four-amino-acid deletion outside the VR2, and another showed no expression of PorA on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Our results reveal that numerous genetic events have occurred in the porA gene of the ET-15 clone in the short time of its epidemic spread. The magnitude of microevolutionary mechanisms available in meningococci and the remarkable genetic flexibility of these bacteria need to be considered in relation to PorA vaccine development.
1986年,在加拿大首次检测到脑膜炎奈瑟菌电泳型(ET)-37复合物中的ET-15克隆,此后它与世界许多地区的脑膜炎球菌病暴发有关。虽然ET-37复合物的大多数菌株是血清亚型P1.5,2,但ET-15菌株的血清亚型测定往往不完整,血清亚型PorA外膜蛋白的两个可变区(VRs)中只有一个或一个都不与单克隆抗体反应。对具有一个或两个未鉴定血清亚型决定簇的ET-15菌株的porA基因进行DNA序列分析,以确定与单克隆抗体缺乏反应的遗传基础。在来自不同地理来源的38株ET-15菌株中鉴定出14种不同的porA等位基因。分别在18、11、2、2和1株分离物中鉴定出与亚型P1.5a,10d、P1.5,2、P1.5,10d、P1.5a,10k和P1.5a,10a相对应的序列。在其余4株均无法分型的菌株中,两株分别在VR1和VR2内有一个终止密码子,而另外两株的porA基因被插入元件IS1301中断。在具有P1.5,2序列的菌株中,一株在VR1和VR2之间有一个终止密码子,一株在VR2外有一个四氨基酸缺失,另一株在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳上未显示PorA表达。我们的结果表明,ET-15克隆的porA基因在其流行传播的短时间内发生了许多遗传事件。在开发PorA疫苗时,需要考虑脑膜炎球菌中可用的微进化机制的程度以及这些细菌显著的遗传灵活性。