PCR反向点杂交技术快速鉴定BALF中分枝杆菌菌种的应用研究

目的探讨PCR-反向点杂交技术(PCR-RDBHA)快速鉴定支气管肺泡灌洗液(bronchoalveolar lavage fluid BALF)标本中分枝杆菌菌种的临床应用价值。方法对临床306例活动性肺结核患者、疑似非结核分枝杆菌感染肺病患者BALF标本采用PCR-反向点杂交技术和传统的改良罗氏培养法进行分枝杆菌菌种鉴定。结果 306例BALF标本PCR反向点杂交技术检测出分枝杆菌127例,阳性率为41.5%(127/306),其中结核分枝杆菌复合群(MTC)108例,占分枝杆菌的85.0%(108/127),非结核分枝杆菌(NTM)19例,占分枝杆菌的15.0%(19/127)。NTM中,6例脓肿分枝杆菌,10例胞内分枝杆菌,2例戈登分枝杆菌,1例瘰疬分枝杆菌;传统的改良罗氏培养法检测出分枝杆菌101例,阳性率33.0%(101/306),经菌种的初步鉴定,其中MTC89例,占分枝杆菌的88.1%(89/101),NTM12例,占分枝杆的11.9%(12/101)。结论 PCR反向点杂交技术能够快速鉴定BALF中分枝杆菌菌种,该方法简便、结果准确,具有良好的临床应用价值。

The application research of PCR-reverse dot blot hybridization assay for rapid identification of Mycobacterium species in BALF

Objective To identify Mycobacterium species in BALF rapidly by polymerase chain reaction-reverse dot blot hy-bridization assay (PCR-RDBHA). Methods 306 cases of BALF from pulmonary tuberculosis patients and nontuberculous My-cobacteria (NTM) lung diseases patients were collected,Mycobacterium species of 360 cases of BALF were simultaneously detected by PCR-RDBHA and Roche medium culture. Results The positive rates of detecting Mycobacterium by PCR-RDBHA were 41. 5%(127/306),108 cases (85.0%) were determind to be M. tuberculosis comples(MTC),19 cases were determind(15.0%) to be NTM which contained 6 cases of M. abscessus,10 cases.of M. intracellulare,2 cases of M. gordonae and 1case of M. scrofulaceum. The positive rates of detecting mycobacterium by culture were 33.0%(101/306),89 cases (88.1%) were determined to be M. tuberculosis comples (MTC),12 cases were determined (11.9%) to be NTM. Conclusion PCR reverse dot blot hybridization technique can quickly identify mycobacterium species in BALF. The method is simple and accurate,and has good clinical application value.