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.
Department of Microbiology, University of Antwerp UIA, Wilrijk, Belgium.
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C
Ovyn
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D
van Strijp
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M
Ieven
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D
Ursi
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van Gemen
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Goossens
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