Detection of M. leprae by reverse transcription- PCR in biopsy specimens from leprosy cases: a preliminary study.

A reverse transcription (RT)-PCR assay targeting 16S rRNA of Mycobacterium leprae has been used to detect M.leprae specific nucleic acids. This study has been initiated to gain experience about detection of RNA from seven biopsy specimens by RT-PCR assay using species- specific primers described earlier. These biopsy specimens were from clinically confirmed and untreated leprosy cases belonging to BB and BL types. The earlier reported method was established in our laboratory. 171 bp fragment by RT-PCR was amplified from 4/7 cases. The positives results by RT-PCR were from the biopsies from fresh or short term treated cases whereas negative results were from specimens from long term treated cases showing clinical features of relapse. DNA targeting PCR (36 KDa) showed positivity in both groups. These results suggest that RT-PCR positivity possibly reflect the presence of viable organisms. Thus as earlier predicted RT-PCR assay may be useful for viability determinations for assessing the response to chemotherapy as well as presence of persisters in relapse cases.

Assignment of the agent of Tyzzer's disease to Clostridium piliforme comb. nov. on the basis of 16S rRNA sequence analysis.

The small-subunit rRNA (16S rRNA) sequence of Tyzzer's bacillus (also known as "Bacillus piliformis") was elucidated by using the polymerase chain reaction followed by reverse transcriptase sequencing. By using maximum-likelihood analysis, a phylogenetic tree was constructed from this and other 16S rRNA sequences available from the first release of the Ribosomal Database Project (G. J. Olsen, R. Overbeek, N. Larsen, T. L. Marsh, M. J. McCaughey, M. A. Maciukenas, W.-M. Kuan, T. J. Macke, Y. Xing, and C. R. Woese, Nucleic Acids Res. 20:2199-2200, 1992). Tyzzer's bacillus grouped with a specific set of anaerobic bacteria, most of which are Clostridium spp. The closest identified relatives are Clostridium coccoides, Clostridium oroticum, Clostridium clostridiiforme, Clostridium symbiosum, and Streptococcus hansenii. Clostridium amino-valericum and "Acetitomaculum ruminis" are also solidly allied with this ensemble. We propose that Tyzzer's bacillus be reclassified as Clostridium piliforme on the basis of its 16S rRNA sequence.