Ozkara H A, Kocagöz T, Ozçelik U, Akçören Z, Göçmen A
Department of Biochemistry, Faculty of Medicine, Hacettepe University, Ankara, Turkey.
Int J Tuberc Lung Dis. 1998 Jun;2(6):451-5.
More than five different primer pairs have been used for the detection of Mycobacterium tuberculosis deoxyribonucleic acid (DNA) with the polymerase chain reaction (PCR).
The sensitivity and specificity of PCR were evaluated using three different primer pairs in the detection of M. tuberculosis in paraffin-embedded tissues.
Thirty-eight tissue specimens from 23 patients were studied. Eighteen samples were obtained from 10 tuberculosis patients, and 20 samples obtained from 13 patients with other diseases were used as negative controls. DNA extracted from paraffin-embedded tissues was used directly for PCR amplification using primers IS1 and IS2 to amplify a 123 base pair (bp) region of IS6110, sjMT3 and sjMTr2 to amplify a 281 bp region of protein antigen b, and INS1 and INS2 to amplify a 245 bp region of IS986. Each amplification was performed double-blinded and repeated three times including positive and negative control samples.
IS1 and IS2 gave a positive result in each of the double samples obtained from eight tuberculosis patients and in the single samples obtained in the two others, sjMT3 and sjMTr2 detected 13 of the 18 tuberculosis samples, and INS1 and INS2 detected only three of the 18.
These results highlight the importance of selecting appropriate primers to obtain high sensitivity in detecting M. tuberculosis in paraffin-embedded tissues by PCR.
聚合酶链反应（PCR）检测结核分枝杆菌脱氧核糖核酸（DNA）时已使用了五种以上不同的引物对。
评估三种不同引物对在石蜡包埋组织中检测结核分枝杆菌时PCR的敏感性和特异性。
研究了23例患者的38个组织标本。从10例结核病患者中获取18个样本，从13例其他疾病患者中获取20个样本作为阴性对照。使用引物IS1和IS2对从石蜡包埋组织中提取的DNA直接进行PCR扩增，以扩增IS6110的123个碱基对（bp）区域，使用sjMT3和sjMTr2扩增蛋白抗原b的281 bp区域，使用INS1和INS2扩增IS986的245 bp区域。每次扩增均采用双盲法进行，并重复三次，包括阳性和阴性对照样本。
IS1和IS2在从8例结核病患者获得的双份样本以及另外2例患者的单份样本中均给出阳性结果，sjMT3和sjMTr2在18份结核病样本中检测到13份，INS1和INS2在18份中仅检测到3份。
这些结果凸显了选择合适引物对于通过PCR在石蜡包埋组织中检测结核分枝杆菌获得高敏感性的重要性。
