Hatta Mochammad, Sultan Andi Rofian, Tandirogang Nataniel
Department of Medical Microbiology, Molecular Biology and Immunology Laboratory for Infectious Diseases, Faculty of Medicine, Hasanuddin University, Jl Perintis Kemerdekaan Km 10 Tamalanrea, Makassar 90245, South Sulawesi, Indonesia.
BMC Res Notes. 2010 Mar 16;3:72. doi: 10.1186/1756-0500-3-72.
Detection of Tuberculosis agent like nontuberculous mycobacteria (NTM) species by culture and microscopic methods remains difficult and time consuming. A fast and reliable diagnosis of tuberculosis would greatly improve the control of the disease. The purpose of this study is to compare the conventional multiplex PCR and multiplex PCR reverse cross blot hybridization assay to culture method in terms of mycobacteria species detection.
Among the 117 positively cultured samples, nontuberculous mycobacteria (NTM) species were found in 9 samples of multiplex PCR reverse cross blot hybridization assay; compared to only 3 NTM species found in our conventional multiplex PCR, and 13 NTM species were successfully identified among 162 negatively cultured samples compared to only 5 NTM species identification in conventional multiplex PCR results.
The sensitivity of the multiplex PCR reverse cross blot hybridization assay comparing to culture method was 86.03%, the specificity is 35.46%, the positive predictive value was 41.94% and the negative predictive value was 82.41%. For conventional multiplex PCR these values are 81.62%, 38.65%, 41.89%, 79.51%, respectively. Furthermore, in terms of mycobacteria species detection, the conventional multiplex PCR was relatively equal compared to the multiplex PCR reverse cross blot hybridization assay, and to be particularly having no significant discrepant results on the identification of Mycobacteria tuberculosis in both methods.
通过培养和显微镜检查方法检测结核分枝杆菌等非结核分枝杆菌（NTM）菌种仍然困难且耗时。快速可靠地诊断结核病将大大改善对该疾病的控制。本研究的目的是在分枝杆菌菌种检测方面，将传统多重聚合酶链反应（PCR）和多重PCR反向杂交印迹法与培养方法进行比较。
在117份培养呈阳性的样本中，多重PCR反向杂交印迹法检测到9份样本中存在非结核分枝杆菌（NTM）菌种；相比之下，我们的传统多重PCR仅检测到3份NTM菌种，在162份培养呈阴性的样本中，成功鉴定出13份NTM菌种，而传统多重PCR结果仅鉴定出5份NTM菌种。
与培养方法相比，多重PCR反向杂交印迹法的灵敏度为86.03%，特异性为35.46%，阳性预测值为41.94%，阴性预测值为82.41%。传统多重PCR的这些值分别为81.62%、38.65%、41.89%、79.51%。此外，在分枝杆菌菌种检测方面，传统多重PCR与多重PCR反向杂交印迹法相对相当，并且在两种方法中对结核分枝杆菌的鉴定没有显著差异结果。
