广东省肇庆市肺炎链球菌分子分型及毒力基因检测

Molecular typing and virulence gene detection of Streptococcus pneumoniae in Zhaoqing,Guangdong

目的了解肺炎链球菌常见血清型及其5种毒力基因的携带情况,为新型疫苗开发研究和提高临床诊断提供基础科学依据。方法选取2016年4月—2018年12月肇庆市第一人民医院住院及门诊患者分离200株肺炎链球菌,采用多重PCR分子分型方法进行血清型分型,再用单因子血清对6A/6B型进行细分。利用PCR方法对肺炎链球菌5种毒力基因(ply、pspA、nanA、psaA、lytA)进行检测,使用SPSS 19.0进行统计学分析。结果200株肺炎链球菌来自男性患者144例,女性患者56例。0~2岁的患者最多(占67.5%),其次是≥50岁患者为16.5%。标本类型以痰为主,占92.0%。多重PCR法分型率为95.0%,血清型以19F(37.5%)、6B(12.0%)、23F(10.0%)、19A(9.0%)、3(6.5%)、14(6%)、15B/15C(5.5%)为主。5种毒力基因检测阳性率分别为ply 93.5%、pspA 85.0%、nanA 88.5%、psaA 90.5%、lytA 93.0%。8株血培养、1株脑脊液培养及19A、3和35A/35C/42三种血清型的5种毒力基因阳性率均为100.0%。血清型14型和5型的5种毒力基因阳性率都较低。结论肇庆市200株肺炎链球菌的血清型以19F、6B、23F、19A、3、14、15B/15C为主,2岁以下的儿童和老年人感染率高。5种毒力基因保守性高,大部分菌株均存在。血清型不同,5种毒力基因的存在也略有不同,可为新型疫苗开发及疾病预防提供有利的依据。

Objective To understand the common serotypes and five virulence genes of Streptococcus pneumoniae,we provide a useful basis for clinical diagnosis and application of new vaccines.Methods Select the 200 strains of Streptococcus pneumoniae which isolated from the First People's Hospital of Zhaoqing from April 2016 to December 2018 for the research objects,serotyping was performed by multiplex polymerase chain reaction(PCR),then serotype 6A/6B was subdivided by single factor serum.Five virulence genes(ply,pspA,nanA,psaA,lytA)of Streptococcus pneumoniae were detected by PCR,used SPSS 19.0 to statistical analysis.Results The male patients(144 cases)more than female patients(56 cases)of the 200 strains of Streptococcus pneumonia.Most of the patients were 0-2 years old,accounting for 67.5%.Over 50 years old(16.5%)patients were the second.The main type of specimens was sputum,accouting for 92.0%.The typing rate of the multiple PCR method was 95.0%,mainly serotypes were 19F(37.5%),6B(12.0%),23F(10.0%),19A(9.0%),3(6.5%),14(6%)and 15B/15C(5.5%).The positive rates of five virulence genes were ply 93.5%,pspA 85.0%,nanA 88.5%,psaA 90.5%and lytA 93.0%.Eight strains of blood culture,one strain of cerebrospinal fluid culture and three serotypes of 19A,3 and 35A/35C/42,the positive rate of five virulence genes was 100.0%.The positive rate of five genes of serotypes 14 and 5 were lower.Conclusion The mainly serotypes of 200 strains of Streptococcus pneumoniae in this area were 19F,6B,23F,19A,3,14,15B/15C,children under two years old and elderly with high infection rate.Most strains have five virulence genes with high conservativeness.The existence of five virulence genes is slightly different with different serotypes,which can provide a favorable basis for the development of new vaccines and disease prevention.