Yamazaki T, Nakamura R M
Department of Bacteriology, National Institute of Health, Tokyo, Japan.
Tuber Lung Dis. 1995 Aug;76(4):330-5. doi: 10.1016/s0962-8479(05)80032-2.
The polymerase chain reaction (PCR) is a rapid and specific method used to amplify a certain DNA fragment. It is applicable to rapid diagnosis of mycobacterial infections. By use of species-specific primers, it is possible to identify mycobacteria by PCR. In this study, a newly constructed primer was tested for specificity for Mycobacterium intracellulare in the PCR.
M. intracellulare is one of the most frequently found bacteria in opportunistic infection in AIDS, and rapid identification of this species is important. The purpose of this study was to construct a primer specific to this species as a suitable tool for identification.
PCR products of M. tuberculosis and M. intracellulare, obtained by using the primers YNP-1 and YNP-2, were sequenced and compared. They showed a difference in the base sequences. A sequence unique to M. intracellulare was used as the primer specific to this species. Various mycobacterial and non-mycobacterial DNAs were used as the primer specific to this species. Various mycobacterial and non-mycobacterial DNAs were used as the template to evaluate the specificity of the newly constructed primers, YNP-7 and YNP-8. Sputum samples were also examined by PCR using the primers.
In total 25 species of culture mycobacterial and non-mycobacterial strains and 76 sputum samples were tested by PCR. Only M. intracellulare DNA was amplified with PCR using the primers YNP-7/8.
The specificity of the newly constructed primers for M. intracellulare was confirmed.
聚合酶链反应（PCR）是一种用于扩增特定DNA片段的快速且特异的方法。它适用于分枝杆菌感染的快速诊断。通过使用种特异性引物，利用PCR可以鉴定分枝杆菌。在本研究中，对新构建的引物在PCR中针对胞内分枝杆菌的特异性进行了检测。
胞内分枝杆菌是艾滋病机会性感染中最常见的细菌之一，快速鉴定该菌种很重要。本研究的目的是构建一种针对该菌种的特异性引物，作为合适的鉴定工具。
对使用引物YNP-1和YNP-2获得的结核分枝杆菌和胞内分枝杆菌的PCR产物进行测序和比较。它们在碱基序列上存在差异。将胞内分枝杆菌特有的一段序列用作该菌种的特异性引物。使用各种分枝杆菌和非分枝杆菌DNA作为模板来评估新构建的引物YNP-7和YNP-8的特异性。还用这些引物通过PCR检测痰液样本。
总共对25种培养的分枝杆菌和非分枝杆菌菌株以及76份痰液样本进行了PCR检测。使用引物YNP-7/8通过PCR仅扩增出了胞内分枝杆菌DNA。
新构建的针对胞内分枝杆菌的引物的特异性得到了证实。
