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  • Title: [Single-tube nested polymerase chain reaction technique in detecting Mycobacterium tuberculosis DNA].
    Author: Ma H, Zhu J, Zhang N.
    Journal: Zhonghua Jie He He Hu Xi Za Zhi; 1998 Jul; 21(7):399-401. PubMed ID: 11326876.
    Abstract:
    OBJECTIVE: To study the sensitivity and specificity of single-tube nested polymerase chain reaction (SN-PCR) technique in detecting Mycobacterium tuberculosis DNA in paraffin-embedded tissues. METHOD: BCG DNA and the paraffin-embedded tissues of 30 cases with typical tuberculous lymphadenitis were examined to detect the IS6110 specific insertion sequences DNA of Mycobacterium tuberculosis complex by general PCR(G-PCR), double-tube nested PCR (DN-PCR) and SN-PCR techniques. RESULT: Fifteen fg or more BCG DNA could show positive results by DN-PCR and SN-PCR techniques, while 480 fg by G-PCR method. The positive rates of G-PCR, DN-PCR, and SN-PCR in detecting Mycobacterium tuberculosis DNA in 30 cases with tuberculous lymphadenitis were 43%, 100%, and 100% respectively, all of which were significantly different (P < 0.01) from that of the acid-fast staining method (10%). Significant differences in the positive rates also existed between G-PCR and DN-PCR, G-PCR and SN-PCR (both P < 0.01), while the positive rates of DN-PCR and SN-PCR were found same. CONCLUSION: The sensitivities of nested PCR techniques in detecting Mycobacterium tuberculosis are significantly higher than that of G-PCR, whereas the sensitivities as well as the specificities of SN-PCR and DN-PCR are the same. Thus SN-PCR seems more practicable.
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