PCR-RDB和SAT与改良罗氏培养法检测临床标本分枝杆菌研究

Detection of Mycobacteriumin clinical specimens by PCR-RDB,SAT and Roche culture

目的对比研究PCR-反向点杂交技术(PCR-RDB)与RNA恒温扩增法(SAT)快速检测临床标本中分枝杆菌菌种的应用价值。方法选取2016年7月-2017年6月医院2 069份活动性肺结核、疑似非结核分枝杆菌感染、肺外结核感染等患者的临床标本,采用PCR-反向点杂交技术进行分枝杆菌菌种检测。同时随机选择临床标本200份行SAT检测;437份行改良罗氏培养法(L-J法)培养和菌种初步鉴定,明确PCR-RDB法临床应用价值。结果 PCR-RDB、SAT法阳性率分别为38.5%(796/2069)、41.5%(83/200),高于L-J法的33.0%(144/437)(P<0.05);PCR-RDB法不仅阳性率高,而且还可以鉴定分枝杆菌菌种,其中检出结核分枝杆菌群(MTC)682份,占分枝杆菌阳性率85.7%(682/796),非结核分枝杆菌(NTM)114份,占分枝杆菌阳性率14.3%(114/796)。结论PCR-RDB法阳性率远高于培养法,略低于SAT法,但PCR-RDB方法简便、结果准确,且能够快速检测临床标本中分枝杆菌菌种,具有良好的临床应用价值。

OBJECTIVE To comparatively study the application value of PCR-reverse dot blot(PCR-RDB)and RNA isothermal amplification(SAT)for rapid detection of Mycobacterium species in clinical specimens.METHODS 2069 cases of clinical specimens from patients with active tuberculosis,suspected non-tuberculous mycobacterium infection,or extrapulmonary tuberculosis infection were collected.The Mycobacteriumspecies in the specimens were detected by PCR reverse dot blot hybridization assay.Meanwhile,200 clinical specimens were randomly selected for SAT examination,and 437 specimens were examined by modified Roche culture(L-J method)and strain identification,in order to determine the clinical application value of the PCR-RDB method.RESULTS The positive rates of detecting Mycobacterium by PCR-RDB and SAT were 38.5%(796/2069)and 41.5%(83/200),respectively,significantly higher than that of the L-J method which was 33.0%(144/437)(P<0.05).The PCR-RDB method had high positive rate,and was able to identify Mycobacteriumspecies.682 cases were determined to be M.tuberculosis complex(MTC),accounting for 85.7%(682/796)in all Mycobacterium-positive specimens,and 114 cases were determined as non-tuberculous Mycobacterium(NTM),accounting for 14.3%(114/796).CONCLUSION The positive rate of PCR-RDB method was far higher than that of the culture method,but slightly lower than that of the SAT method.The PCR-RDB method is simple and accurate,and can quickly identify Mycobacteriumspecies in clinical specimens.Therefore,this method has good clinical application value.