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目的探讨反向线性点杂交技术在肺炎链球菌分型中的应用。方法对临床诊断为肺炎的16例5岁以下患儿,从呼吸道吸取物、血和胸水培养分离获得的30株肺炎链球菌菌株应用反向线性点杂交技术进行分型,并与传统的血清学分型方法-夹膜肿胀实验进行比较。结果反向线性点杂交技术和血清学方法对30株临床分离获得的肺炎链球菌菌株的分型结果完全一致,其分布情况为:血清型19A(n=9),14(n=5),23F(n=4),19F(n=4),6B(n=3),7F(n=2),15B(n=2),9V(n=1)。16例肺炎患儿的肺炎链球菌血清型分布情况为血清型19A、14、23F及19F各来自3例患儿,血清型6B、7F、15B及19V各来自1例患儿。其中10例(62.5%)肺炎患儿分离获得的肺炎链球菌菌株涵盖在7价肺炎链球菌结合疫苗中,所有患儿获得的菌株均包含在23价多糖疫苗中。结论对于肺炎链球菌,反向线性点杂交技术是一种可靠的分子生物学分型方法;在本组研究中,7价肺炎链球菌结合疫苗的涵盖率为62.5%,所有菌株均包含在23价多糖疫苗中。
Objective To investigate the application of reverse linear dot blot technique in the typing of Streptococcus pneumoniae. Methods Thirty cases of Streptococcus pneumoniae strains isolated from respiratory tract aspirates, blood and pleural effusion isolated from 16 children under 5 years of age clinically diagnosed as pneumonia were genotyped by reverse linear dot blot hybridization and compared with traditional serological tests Type method - jacketed swelling experiments were compared. Results The results of reverse linear dot blot hybridization and serological methods were identical to those of 30 strains of S. pneumoniae isolated clinically. The distribution of serotype 19A (n = 9), 14 (n = 5) 23F (n = 4), 19F (n = 4), 6B (n = 3), 7F (n = 2), 15B (n = 2), 9V (n = 1). The distribution of S. pneumoniae serotypes in 16 children with pneumonia was serotype 19A, 14,23F and 19F each from 3 children, and one from each of serotypes 6B, 7F, 15B and 19V. Among them, 10 strains (62.5%) of pneumococcal strains isolated from children with pneumonia were covered in the 7-valent pneumococcal conjugate vaccine, and all the strains obtained from children were included in the 23-valent polysaccharide vaccine. Conclusions For Streptococcus pneumoniae, reverse linear dot blot is a reliable molecular biological typing method. In this study, the coverage of the 7-valent pneumococcal conjugate vaccine was 62.5%, and all strains were included in the 23-valent Polysaccharide vaccine.