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本研究以结核杆菌染色体中重复出现的长度为１２３个碱基对（ｂｐ）的ＤＮＡ片段作为聚合酶链反应（ＰＣＲ）扩增的模板，合成一对寡核苷酸引物。痰液标本先经三羟甲基氨基甲烷（Ｔｒｉｓ），乙二胺四乙酸二钠（ＥＤＴＡ－２Ｎａ）和三硝基甲苯（Ｔｒｉｔｏｎｘ－１００）加热、震荡处理，裂解结核杆菌，再作ＰＣＲ扩增和琼脂糖凝胶电泳。同时，与痰液直接涂片、抗酸染色镜检抗酸染色阳性杆菌作比较。用该ＰＣＲ检测的检出率为９１．４％（３２／３５），而涂片抗酸染色的检出率仅为３４．３％（１２／３５）。两者相比有非常显著性差异（ｘ２＝２４．４７６，Ｐ＜０．００１）。由于改进了ＰＣＲ前痰液标本的处理方法，使整个ＰＣＲ检测过程所需时间缩短至９ｈ。我们认为该ＰＣＲ检测结核杆菌是特异、敏感和快速的，可在临床实验室应用。
In this study, a DNA fragment of 123 base pairs (bp) repeated in the chromosome of Mycobacterium tuberculosis was used as template for polymerase chain reaction (PCR) amplification to synthesize a pair of oligonucleotide primers. The sputum samples were first heated by Tris, EDTA-2Na and Tritonx-100, shock-treated to lyse Mycobacterium tuberculosis and then PCR-amplified Adipose and agarose gel electrophoresis. At the same time, direct smear with sputum, acid-fast staining antimicrobial acid-positive bacilli for comparison. The detection rate by this PCR was 91.4% (32/35), while the smear acid-fast staining detection rate was only 34.3% (12/35). There was a significant difference between the two groups (x2 = 24.476, P <0.001). Due to the improved handling of sputum samples prior to PCR, the time required for the entire PCR assay was shortened to 9 hours. We think this PCR detection of Mycobacterium tuberculosis is specific, sensitive and rapid and can be used in clinical laboratories.