反向斑点杂交技术鉴定分枝杆菌菌种的应用评价

Evaluation of reverse blot hybridization assay for identification of Mycobacterium species

目的评估反向斑点杂交技术(reverse blot hybridization assay,REBA)检测结核分枝杆菌(MTB)与非结核分枝杆菌(NTM)在临床诊断中的应用价值。方法采用对硝基苯甲酸(PNB)初筛、REAB检测及核酸测序方法检测河南省胸科医院临床不同结核病患者痰标本罗氏培养分离出的分枝杆菌78株,探讨3种方法鉴定结核分枝杆菌复合群与非结核分枝杆菌结果的一致性。结果 REBA检测78株分枝杆菌菌型,73株属结核分枝杆菌复合群,4株为非结核分枝杆菌,1株鉴别为混合菌;与核酸测序检测结果符合率为98.7%(77/78),一致性极好(Kappa=0.99);验证REBA与PNB初筛结果显示,符合率为94.9%(74/78),两法结果一致性好(Kappa=0.95)。结论利用REBA对临床分离的分枝杆菌进行菌种鉴定是一种快速、简便和准确的方法,可满足早诊断,早治疗的临床需要。

Objective To evaluate clinical effectiveness of reverse blot hybridization assay（ REBA） for identification of Mycobacterium species and nontuberculosis Mycobacteria. Methods 78 clinical Mycobacterium isolates collected from Henan Chest Hospital were detected by PNB method,reverse blot hybridization assay and DNA sequencing,respectively,so as to explore the coincidence of the detection results of Mycobacterium tuberculosis and nontuberculosis Mycobacteria with the above three methods.Results There were 78 strains of Mycobacteria detected by REBA 73 strains were complex Mycobacterium tuberculosis,4 strains were non-mycobacterium and 1 strain was mixed bacteria. The coincidence rate with nucleic acid sequencing was 98. 7%（ 77/78） and coincidence was good（ Kappa = 0. 99）. The results of primary screening of REBA and PNB showed that the coincidence rate was 94. 9%（ 74/78）. The agreement between the two methods was good（ Kappa = 0. 95）. Conclusion The use of REBA for clinical identification of Mycobacteria is a rapid,simple and accurate method,which can meet the clinical needs of early diagnosis and early treatment.