应用缺口-连接酶链反应检测新生儿沙眼衣原体感染  

Development of a gap ligase chain reaction for detection of Chlamydia trachomatis in newborn infants

作　　者：韦红[1] 吴仕孝[1] 余加林[1] 杨军[2] 刘官信[1] 

机构地区：[1]重庆儿童医院新生儿科,重庆400014 [2]第一军医大学附属珠江医院儿科

出　　处：《中华儿科杂志》2003年第8期578-581,共4页Chinese Journal of Pediatrics

基　　金：国家自然科学基金资助项目 (3 9670 766)

摘　　要：目的　建立缺口 连接酶链反应 (gapligasechainreaction ,G LCR)诊断新生儿沙眼衣原体(Chlamydiatrachomatis)感染。方法　根据沙眼衣原体主要外膜蛋白基因设计两对探针 ,应用LCR技术扩增衣原体标准株DNA ,并用于检测 32 8例新生儿肺炎鼻咽拭子标本中的沙眼衣原体 ,在敏感性、特异性方面与传统的细胞培养法进行比较。结果　G LCR可检测出两个衣原体原体 ;可扩增 5种沙眼衣原体标准株DNA ,不扩增鹦鹉热衣原体及其他细菌DNA ,显示出很高的特异性。 32 8例鼻咽拭子标本中 ,G LCR扩增检出 6 7例阳性 ,培养检出 6 0例。经结果不一致分析后 ,共检出 6 9例阳性 ,阳性率为2 1%。以扩大标准为金标准判断 ,G LCR的敏感性、特异性、阳性预测值和阴性预测值分别为 98 6 %、10 0 %、10 0 %和 99 6 % ,而培养则分别为 86 9%、10 0 %、10 0 %和 96 6 %。结论　G LCR检测新生儿沙眼衣原体感染具有较高的敏感性和特异性。G LCR是一种有应用前景的核酸扩增方法。Objective To establish a gap ligase chain reaction (G-LCR) assay for the detection of Chlamydia trachomatis (Ct) in neonates with pneumonia. Methods A G-LCR DNA amplification assay that targeted the outer major membrane protein gene (omp1) of Ct was developed to detect Ct. The sensitivity and specificity of the G-LCR test was examined by the use of highly purified elementary bodies (EBs). Nasopharyngeal swabs taken from 328 neonates with pneumonia were analyzed by Gap-LCR and cell culture. Results The detection limit of G-LCR was 2 EBs . G-LCR could detect five species of Ct and was not cross-reacted with C psittaci and other bacteria. The prevalence of Ct in 328 neonates with pneumonia, using an expanded gold standard of a positive cell culture or two confirmed positive non-culture tests, was 21% (69/328). After analysis of discrepant results, the sensitivity, specificity, and positive and negative predictive values for the G-LCR were 98.6%, 100%, 100% and 99.6%, respectively; whereas those for culture were 86.9%, 100%, 100% and 96.6%, respectively. Conclusion This study demonstrated that the G-LCR was a highly sensitive nonculture technique and good alternative test for the detection of chlamydial infections.

关 键 词：缺口-连接酶链反应 检测 新生儿 沙眼衣原体感染 核酸扩增法 

分 类 号：R722.13[医药卫生—儿科] R446.5[医药卫生—临床医学]