Detection of Mycobacterium leprae by three-primer PCR.
Recently, polymerase chain reaction has been introduced for the species-specific assessment of Mycobacterium leprae (1). To avoid Southern blotting techniques using radioactively labelled oligonucleotide probes, the aim of this study was to establish a three primer-based single-step PCR technique. Using primers designed for this purpose we amplified a part of the gene encoding for the 16S ribosomal RNA of slowly growing mycobacteria. Due to the species-specific antisense primer a second, smaller fragment specific for M. leprae was amplified. Our results show that the employment of a second antisense primer in the PCR may be a substitution for Southern blot hybridization.
Richter
E
E
Department of Immunology and Cell Biology, Forschungsinstitut Borstel, Germany.
Duchrow
M
M
Schlüter
C
C
Hahn
M
M
Flad
H D
HD
Gerdes
J
J
D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D005069:Evaluation Studies as Topic; D006801:Humans; D007918:Leprosy / Q000175:diagnosis; D008969:Molecular Sequence Data; D009166:Mycobacterium leprae / Q000235:genetics* / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D013045:Species Specificity
D017931:DNA Primers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S