多重聚合酶链反应与荚膜肿胀试验对肺炎链球菌血清分型的比较研究

Serotyping of Streptococcus pneumoniae by multiplex polymerase chain reaction and capsular swelling test

为评估多重聚合酶链反应(PCR)对肺炎链球菌血清分型的可行性,分别采用多重PCR和荚膜肿胀试验对568株肺炎链球菌进行血清分型,并对分型结果进行比较分析。结果显示,568株肺炎链球菌中,213株通过荚膜肿胀试验分出16个血清群,主要有血清群19(23.1%,131/568)、6(5.3%,30/568)、23(1.6%,9/568)、14(1.4%,8/568)、9(1.1%,6/568)、15(1.1%,6/568)等,分型率为37.5%(213/568);356株通过多重PCR分出21个血清群,主要有血清群19(27.8%,158/568)、23(8.5%,48/568)、6(7.4%,42/568)、14(4.4%,25/568)、3(4.2%,24/568)、15(3.5%,20/568)等,分型率为62.7%(356/568)。荚膜肿胀试验鉴定出血清群4和18,但多重PCR未能鉴定;多重PCR鉴定出血清群5、12、35、16、17和22,但荚膜肿胀试验未能鉴定。多重PCR与荚膜肿胀试验对19F、19A血清型的鉴定无显著差异。结果提示,这2种方法对肺炎链球菌血清分型结果有差别,多重PCR的分型率高于荚膜肿胀试验。对来源复杂的标本进行肺炎链球菌血清分型,2种方法可相互补充,以提高分型率。

The main purpose of the research is to evaluate the application feasibility of multiplex polymerase chain reaction （PCR） in serotyping of Streptococcus pneumoniae .A total of 568 strains were subjected to multiplex PCR and capsular swelling test respectively .The results demonstrated that 37 .5% of the strains （213/568） could be successfully serotyped by capsular swelling test into 16 different serotypes ,including serotypes 19 （23 .1% ,131/568） ,6 （5 .3% ,30/568） ,23 （1 .6% ,9/568） ,14 （1 .4% ,8/568） ,9 （1 .1% ,6/568） and 15 （1 .1% ,6/568 ）;while 62 .7% of the strains （356/568 ） could be successfully serotyped by multiplex PCR into 21 different serotypes ,including serotypes 19 （27 .8% ,158/568） ,23 （8 .5% ,48/568） , 6 （7 .4% ,42/568） ,14 （4 .4% ,25/568） ,3 （4 .2% ,24/568） and 15 （3 .5% ,20/568） .The multiplex PCR could not identify serotypes 4 and 18 which could be identified by capsular swelling test ,while the capsular swelling test could not identify serotypes 5 ,12 ,35 ,16 ,17 and 22 which could be successfully identified by multiplex PCR .There was no significant difference between the two methods to identify serotypes 19F and 19A .The results suggest that there is significant difference between the two methods in serotyping of Streptococcus pneumoniae .Multiplex PCR is more effective than the conventional capsular swelling test , especially for those specimens collected from complicated sources .The combination of the two methods can improve the serotyping efficiency of Streptococcus pneumoniae .