摘要
目的了解天津市应用聚合酶链反应(PCR)方法对百日咳病例的诊断效果,探讨确诊病例分子流行病学特点。方法选取天津市医院及社区监测的百日咳疑似病例110例,采集鼻咽拭子标本,应用PCR法检测百日咳杆菌的筛选及验证基因;同时采用酶联免疫吸附试验(ELISA)法检测血清中特异性百日咳毒素IgG(PT-IgG)抗体;采用PCR扩增确诊病例百日咳毒素(PT)S1亚单位基因,进行序列测定和分析。结果 110例百日咳疑似病例中,PCR检测阳性率60.00%;ELISA检测94例,PT-IgG抗体阳性率42.55%,2种方法检测阳性率差异有统计学意义(χ2=6.181,P=0.013);<1岁病例PCR检测阳性率高于其他年龄段;19株百日咳杆菌基因测序的亲缘性关系密切,核苷酸同源性为99.88%;在PT区域S1亚单位第218位氨基酸发生变异,蛋氨酸(Met)变异为异亮氨酸(Iso)。结论天津市百日咳杆菌基因的亲缘性关系密切;应用PCR检测鼻咽拭子进行百日咳病例确诊,方法敏感,操作易行。
Objective To examine the application of polymerase chain reaction（PCR） method in diagnosis of pertussis and to explore molecular epidemiological features of pertussis.Methods Totally 110 suspected pertussis cases were selected and their nasopharyngeal swabs were sampled at hospitals or community monitoring settings in Tianjin.Genetic screening of Bordetella pertussis gene was conducted with PCR and ELISA was used to detect specific pertussis toxin（PT）-IgG antibody.PT S1 subunit gene region in pertussis DNA positive samples was sequenced and analyzed.Results The PCR positive rate was 60% in 110 suspected pertussis cases and the PT-IgG antibody positive rate was 42.55% in 94 suspected pertussis cases.PCR positive rate was significantly higher than that of PT-IgG antibody（χ2=6.181,P=0.013）.The PCR detection rate was significantly higher in the cases less than one year old than in the cases of other age groups.All DNA gene 19 Bordetella pertussis strains were closely related with a nucleotide homology of 99.88%.In the S1 subunit of PT region,amino acid 218 mutated from methionine（Met） to isoleucine（Iso）（Met218 → Iso218）.Conclusion PCR detection of clinical nasopharyngeal swab sample for diagnosis of pertussis is sensitive and convenient.The Bordeteaal pertussis prevalent in Tianjing is closely related in gene homology.
出处
《中国公共卫生》
CAS
CSCD
北大核心
2011年第8期987-989,共3页
Chinese Journal of Public Health
基金
天津市科技支撑重大项目(07SYSYSF05100)
关键词
百日咳杆菌
聚合酶链反应
序列分析
Bordetella pertussis
polymerase chain reaction
sequence analysis
