Ovyn C, van Strijp D, Ieven M, Ursi D, van Gemen B, Goossens H
Department of Microbiology, University of Antwerp UIA, Wilrijk, Belgium.
Mol Cell Probes. 1996 Oct;10(5):319-24. doi: 10.1006/mcpr.1996.0043.
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.
基于核酸序列的扩增技术（NASBA）是一种用于核酸的等温扩增技术，曾被用于对24株肺炎支原体菌株进行分型。从支原体属的16S rRNA序列比对中选择了一组引物。测定了肺炎支原体菌株M15/83（1型）和FH（2型）的（-）RNA扩增子的核苷酸序列，结果显示这两种类型在16S rRNA水平上存在一个位点差异。基于这一结果，构建了两种类型特异性探针。在酶联凝胶分析（ELGA）中，这些探针与24株肺炎支原体菌株的扩增核酸在溶液中进行杂交。基于NASBA的分型结果与通过其他分型方法将这24株肺炎支原体菌株分为两种类型的分类结果一致，证实了该技术的可靠性。
