PCR-荧光探针法检测临床痰标本结核分枝杆菌的可靠性研究  

Study on the reliability of detecting Mycobacteria in sputum specimens by PCR-fluorescent probe

作　　者：梁建琴[1] 高华方[2] 李洪敏[1] 赵智贤[2] 张广宇[1] 王金河[1] 王甦民[3] 刘佳文 

机构地区：[1]解放军第三〇九医院结核病研究所,北京100091 [2]博奥生物有限公司暨生物芯片北京国家工程研究中心 [3]北京结核病控制研究所 [4]北京市老年病医院检验科

出　　处：《中国防痨杂志》2012年第5期271-274,共4页Chinese Journal of Antituberculosis

基　　金：国家"十一五"科技重大专项(2008ZX10003-001)

摘　　要：目的评价PCR-荧光探针法快速检测Mtb和非结核分枝杆菌(NTM)的临床应用价值。方法应用分枝杆菌核酸检测试剂对1015例痰标本和56株临床分离株进行检测,与Mtb核酸扩增(PCR)荧光(目前临床上认可惟一同类产品)检测结果进行比较。采用SPSS 11.5统计软件进行数据统计处理,采用χ2检验,以P<0.05为差异有统计学意义。结果PCR-荧光探针法检测痰标本分枝杆菌灵敏度50.3%(373/741),特异度98.9%(271/274);菌阳肺结核标本检测灵敏度97.0%(257/265),菌阴肺结核标本检测灵敏度24.4%(116/476);1015份痰标本中检测出11例非结核分枝杆菌(NTM)核酸阳性标本(占1.1%),经16SrDNA测序确认,准确率100.0%。随机数字量表法抽取138例标本重复进行第2次检测,符合率100.0%。对56株临床分离株(1株Mtb临床分离株,55株NTM临床分离株)检测,准确率100.0%。对临床需要的226份标本同时与达安公司试剂盒检测结果比较,总体符合率95.1%(215/226),经统计学分析,两者之间差异无统计学意义(χ2=2.98,P=0.226)。结论PCR-荧光探针法可快速检测和区分痰标本Mtb与NTM,具有临床推广应用价值。Objective To evaluate the clinical application value of detecting the Mycobacterium tuberculosis and non-tuberculous mycobacteria（NTM） by PCR-fluorescent probe. Methods 1015 sputum specimens and 56 clinical isolates were studied using PCR-fluorescent probe of Capital Bio Corporation.And the results were compared with that of Mycobacterium tuberculosis detection kit of Daan Company. Results The sensitivity and specificity of detecting Mycobacteria in the sputum specimens by PCR-fluorescent probe were 50.3%（373/741） and 98.9%（271/274） respectively,in which the sensitivities for smear-positive and smear-negative pulmonary tuberculosis were 97.0%（257/265） and 24.4%（116/476） respectively.Of 1015 sputum specimens,11（1.1%）were detected as NTM,Using 16S rDNA sequencing as control,the accuracy rate was 100.0%.138 sputum specimens were detected repeatedly,the coincidence rate was 100.0%.Of 56 clinical isolates,1 strain was detected as Mycobacterium tuberculosis and 55 were detected as NTM,the accuracy rate was also 100.0%.The total coincidence rate of 226 sputum specimens was 95.1%（215/226） compared with Mycobacterium tuberculosis detection kit of Daan Company.There were no significant difference between two methods（χ2=2.98,P=0.226）. Conclusion PCR-fluorescent probe can rapidly detect and differentiate Mycobacterium tuberculosis and NTM,have the value of clinical application.

关 键 词：分枝杆菌 结核 分枝杆菌属 聚合酶链反应 荧光染料 痰 

分 类 号：R440[医药卫生—诊断学]