PCR直接检测皮肤分枝杆菌感染的研究  

Study of PCR for direct detection and identification of mycobacteria in skin specimens

作　　者：王洪生[1] 李晓杰[1] 吴勤学[1] 崔盘根[1] 刘训荃[1] 

机构地区：[1]中国医学科学院,中国协和医科大学皮肤病研究所,南京210042

出　　处：《中华皮肤科杂志》2006年第10期593-595,共3页Chinese Journal of Dermatology

摘　　要：目的探讨PCR直接检测皮肤分枝杆菌感染作为诊断的可能性。方法PCR检测方法及培养方法(L(?)wenstein-Jensen培养基)比较分枝杆菌纯菌悬液、模拟皮肤分枝杆菌感染标本前处理前后、疑似皮肤分枝杆菌感染的临床标本前处理前后的检测结果。结果分枝杆菌纯菌悬液的PCR法和培养法敏感性皆为1×10~2个菌细胞/mL。模拟临床标本在经过前处理后在浓度为1×10~2个菌细胞/mL的数量级上PCR检测阳性率为60%,而未经过前处理的模拟临床标本在此数量级上PCR检测阳性率为0,培养方法的阳性率为100%,但在浓度为1×10~3个菌细胞/mL数量级上PCR检测阳性率为100%。37例临床疑似皮肤分枝杆菌感染标本经前处理后,PCR检测7例阳性,而未经前处理的标本同法检测2例阳性,但培养方法可检出9例阳性。结论临床皮肤标本经前处理后,PCR检测的敏感性已接近80%,但能明显缩短检测时间。Objective To study the possibility of direct detection and identification of mycobacteria by PCR in skin specimens. Methods The comparison of PCR with in vitro culture of M. smegmatis was conducted at three levels. First, serial dilutions of M. smegmatis DNA was amplified with primers aiming at hsp65 gene to testify the sensitivity of PCR for mycobacterial detection, and the same dilutions of suspension of M. smegmatis was inoculated in I^wenstein-Jensen medium to assessed the sensitivity of in vitro culture. Secondly, the results of detecting simulant clinical specimens by PCR with or without pretreatment were compared with those by in vitro culture. Thirdly, the results of detecting 37 clinical skin specimens suspected to be infected with mycobacteria by PCR with or without pretreatment were compared with those by in vitro culture. Results The sensitivity of both PCR and in vitro culture for detection of serial dilutions of bacterial suspension of M smegmatis was 1 ×10^2 cells/mL. The detection rates of PCR for simulant clinical skin specimen at 1 ×10^2 cells/mL with and without pretreatment were 60% and 0 respectively, and the detection rates of in vitro culture were 80% and 100%, respectively. The detection rate of PCR was 100% for simulant clinical skin specimens at 1 ×10^3 cells/mL. From the 37 clinical skin specimens, 7 and 2 positive cases were detected by PCR with and without pretreatment respectively, however, 9 positive cases were detected by in vitro culture. Conclusion With the pretreatment of clinical skin specimens, the sensitivity of PCR approximates 80% and the time needed for detection is obviously shortened.

关 键 词：分支杆菌属 聚合酶链反应 

分 类 号：R450[医药卫生—治疗学]