Typing of Mycoplasma pneumoniae by nucleic acid sequence-based amplification, NASBA.
Nucleic acid sequence-based amplification, NASBA, is an isothermal amplification technique for nucleic acids and was used for typing a collection of 24 Mycoplasma pneumoniae strains. A set of primers was chosen from the 16S rRNA sequence alignment of Mycoplasma species. The nucleotide sequences of the (-)RNA amplicons were determined for M. pneumoniae strains M15/83 (type 1) and FH (type 2), and revealed a one-point difference at the 16S rRNA level between the two types. Based on this result, two type-specific probes were constructed. The probes were hybridized in solution with the amplified nucleic acids of 24 M. pneumoniae strains in an enzyme-linked gel assay (ELGA). The results obtained by NASBA-based typing are in agreement with the classification of the 24 M. pneumoniae strains into two types by other typing methods, confirming the reliability of this technique.
Ovyn
C
C
Department of Microbiology, University of Antwerp UIA, Wilrijk, Belgium.
van Strijp
D
D
Ieven
M
M
Ursi
D
D
van Gemen
B
B
Goossens
H
H
D015373:Bacterial Typing Techniques; D017931:DNA Primers / Q000235:genetics*; D005784:Gene Amplification; D015336:Molecular Probe Techniques; D009177:Mycoplasma pneumoniae / Q000145:classification* / Q000235:genetics; D012329:RNA, Bacterial / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000235:genetics; D012680:Sensitivity and Specificity; D016415:Sequence Alignment; D017423:Sequence Analysis, RNA
D017931:DNA Primers; D012329:RNA, Bacterial; D012336:RNA, Ribosomal, 16S