Molecular typing of Neisseria gonorrhoeae by restriction fragment length polymorphisms

Abstract

Objective: To characterise Neisseria gonorrhoeae isolates by restriction fragment length polymorphisms (RFLPs) in ribosomal RNA genes. Design: 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. Specimens: Forty three randomly collected isolates from Bangkok (27 isolates) and Singapore (16 isolates) were studied. Results: 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. Conclusion: 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.