Poh C L, Khng H P, Lim C K, Loh G K
Department of Microbiology, National University of Singapore.
Genitourin Med. 1992 Apr;68(2):106-10. doi: 10.1136/sti.68.2.106.
To characterise Neisseria gonorrhoeae isolates by restriction fragment length polymorphisms (RFLPs) in ribosomal RNA genes.
Generation of RFLP patterns by HincII restriction of rRNA genes followed by hybridisation with a non-radioactive labelled broad spectrum 16 + 23S rRNA gene probe. This typing method was developed and compared with MAb based serotyping.
Forty three randomly collected isolates from Bangkok (27 isolates) and Singapore (16 isolates) were studied.
The RFLP patterns generated were reproducible and highly discriminatory between strains. Analysis of RFLPs produced by HincII restriction of rRNA genes established 9 patterns amongst the 43 isolates examined. Strains present within a common serovar could be further subdivided by RFLP typing. Identical RFLP patterns were found in some strains that belonged to various serovars.
RFLP typing based on heterogeneities of rRNA gene restriction patterns could be advantageously used to complement monoclonal antibody based serotyping for further subdivision of serovars. Higher sensitivity of this combined approach would enable better differentiation of strains in epidemiological studies.
通过核糖体RNA基因的限制性片段长度多态性(RFLP)对淋病奈瑟菌分离株进行特征分析。
通过用HincII对rRNA基因进行限制性内切,然后与非放射性标记的广谱16 + 23S rRNA基因探针杂交来产生RFLP图谱。开发了这种分型方法并与基于单克隆抗体的血清分型进行比较。
研究了从曼谷随机收集的43株分离株(27株)和新加坡的分离株(16株)。
产生的RFLP图谱具有可重复性,并且在菌株之间具有高度鉴别性。对rRNA基因进行HincII限制性内切产生的RFLP分析在检测的43株分离株中确定了9种图谱。同一血清型中的菌株可以通过RFLP分型进一步细分。在一些属于不同血清型的菌株中发现了相同的RFLP图谱。
基于rRNA基因限制性图谱异质性的RFLP分型可有利地用于补充基于单克隆抗体的血清分型,以便对血清型进行进一步细分。这种联合方法的更高灵敏度将使在流行病学研究中更好地区分菌株成为可能。