[Detection of Mycobacterium tuberculosis using a nested-primer gene amplification assay].
A nested-primer gene amplification assay (NPGAA) was established by using two pairs of primers, an outside pair of primers and an inside pair of primers, sequences of which were from M. tuberculosis(Mtb) gro EL genes. The outside pair of primers should amplify a 576-bp piece of the Mtb gro EL gene that contains sites for the inside pair of primers, which should amplify a 344-bp piece. The results shows NPGAA'S detectable limitation was 1 organism/ml and no amplification products were produced from DNAs of other mycobacterium species tested in this study. For detecting Mtb, the entire NPGAA, from sample preparation to data analysis, can be completed within 6-8 hours. When identifying AFB cultures isolated from lesions of skin diseases, the same positive size patterns were obtained.
Wu
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QX
Institute of Dermatology, CAMS&PUMC, Nanjing.
Li
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XY
Wei
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WH
D001483:Base Sequence; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis*; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000302:isolation & purification; D021141:Nucleic Acid Amplification Techniques; D016133:Polymerase Chain Reaction / Q000379:methods
D017931:DNA Primers; D004269:DNA, Bacterial