| 摘要 |
Detection of mycobacteria and specific detection of both the Mycobacterium tuberculosis complex and M. avium comprises: (a) extracting microbial DNA; (b) amplifying at least one fragment of the 16S rRNA gene, using a primer pair; (c) detecting the genus-specific region of the amplicon with a pair of labeled hybridization probes; and (d) detecting the species-specific part of the amplicon using separate pairs of labeled probes. Detection of mycobacteria and specific detection of both the Mycobacterium tuberculosis complex and M. avium in clinical material comprises: (a) extracting microbial DNA from the material; (b) amplifying at least one fragment of the 16S rRNA gene, using a primer pair comprising a 21 nucleotide (nt) sequence (1) and 20 nt sequence (5), or two primer pairs comprising (2) and (3), both 18 nt, and (4), of 18 nt, and (5); (c) detecting the genus-specific region of the amplicon with a pair of labeled hybridization probes, i.e. (10) of 23 nt and (11) of 18 nt; and (d) detecting the species-specific part of the amplicon using separate pairs of labeled probes: (6) of 20 nt and (7) of 22 nt for M. tuberculosis and (8) of 20 nt and (9) of 21 nt for M. avium, or their complements. The analysis in (c) and (d) is by analysis of the melting curve and all oligonucleotide sequences are reproduced. Independent claims are also included for the following: (1) The oligonucleotide pairs (2)/(3), (6)/(7), (8)/(9) and (10)/(11), also primer (4); (2) Synthetic plasmid, useful as internal control for amplification and detection of mycobacterial 16S rRNA genes, containing modified sequences for the genus-specific region III of the gene; and (3) Diagnostic kit for the new process. |
