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目的研究脑膜炎奈瑟菌(Neisseria Meningitides,Nm)基因组中串联重复序列(Variable Number of Tandem Repeats Sequences,VNTRs)与血清分群(A和C群)之间的关系。方法采用Nm血清抗体玻片凝集法对Nm菌株进行血清学鉴定;选择4对引物,应用聚合酶链反应(Polymerase Chain Reaction,PCR)扩增Nm基因组中4个位点的VNTR01～04,使用非加权配对算术平均法,应用统计学软件分析VNTRs拷贝(Copy)数与血清群之间的关系;以编码半乳糖转移酶基因中的VNTR01位点之后一段核苷酸序列为特异引物,对Nm进行PCR扩增。结果118株Nm菌株中,38株为A群,80株为C群,聚类分析可将118株菌分为5个组(I、II、III、IV、V)。95.8%(113/118)的菌株位于Ⅰ、Ⅱ组内,Ⅰ组中97.3%(36/37)的菌株为A群,Ⅱ组中98.7%(75/76)的菌株为C群。用两对特异引物对118株菌株进行PCR扩增,以PCR产物有或无做为判断结果,结果与血清分群有96.6%(114/118)的一致性。结论VNTRs拷贝数与血清群之间有较好的相关性,编码半乳糖转移酶基因中变异的核苷酸序列可以做为鉴别A、C群菌株的一种核酸标识。
Objective To study the relationship between Variable Number of Tandem Repeats Sequences (VNTRs) and serogroups (groups A and C) in Neisseria meningitides (Nm) genome. Methods Nm strains were identified by Nm serum antibody agglutination method. Four pairs of primers were selected and VNTR01 ~ 04 of Nm genome was amplified by Polymerase Chain Reaction (PCR) Weighted pairwise arithmetic average method, statistical software was used to analyze the relationship between the number of VNTRs copies and serogroups. A nucleotide sequence encoding the VNTR01 site in the galactosyltransferase gene was used as a specific primer to conduct Nm PCR amplification. Results Among the 118 Nm strains, 38 were Group A and 80 were Group C, and 118 were divided into 5 groups (I, II, III, IV, V) by cluster analysis. 95.8% (113/118) of the isolates were in group Ⅰ and Ⅱ, 97.3% (36/37) in group Ⅰ were group A, and 98.7% (75/76) in group Ⅱ were group C. 118 pairs of specific primers were used to amplify 118 strains of PCR products with or without PCR products as a result of judgment, the result was 96.6% (114/118) consistency with the serum grouping. Conclusion There is a good correlation between VNTRs copy number and serogroups. The nucleotide sequence encoding the mutation in galactosyltransferase gene can be used as a nucleic acid marker to identify group A and C strains.