Typing of Chlamydia trachomatis by restriction endonuclease analysis of the amplified major outer membrane protein gene

Abstract

A procedure was developed for characterization of Chlamydia trachomatis strains by using restriction endonuclease analysis of amplified genes of the major outer membrane protein (MOMP). Reference strains of the 15 serovars (A through K and Li through L3) and clinical isolates were tested. The nucleotide sequences of the MOMP genes of each of the 15 serovars were arbitrarily constructed by using the sequences of the four variable domains known for each serovar and the constant domains of serovar Li. Computer analysis of these sequences indicated that two restriction digestions performed in parallel, one with AluI and the other with HpaII, followed by Hinfl and EcoRI, would allow the theoretical differentiation of 13 serovars. Serovars Ba and Li presented the same theoretical restriction profile. Our typing method consisted of polymerase chain reaction amplification of a fragment of about 1,200 bp of the MOMP gene, followed by restriction endonuclease digestion with the aforementioned enzymes. From the 15 serovars, we obtained 14 different patterns; 13 profiles were serovar specific, while serovars B and Ba presented the same pattern. Application of this typing method to C. trachomatis strains isolated from clinical material gave the same results as the immunotyping method for 14 of 17 strains. Furthermore, restriction endonuclease analysis detected differences within a serovar. This method seems to be promising for epidemiological studies.

Analysis of Chlamydia trachomatis serovars in endocervical specimens derived from pregnant Japanese women

“…The VSs of MOMP are highly immunogenic and are thought to have a role in protective immunity [25,26]. Within the omp 1 VS regions, nucleotide substitution almost invariably results in a di¡erence in amino acid se- Rodriguez et al [5] chose to digest the ampli¢ed 1.2-kbp DNA fragments separately with AluI and HpaII-EcoRI-HinfI in two steps and distinguished 13 of 15 serovars of C. trachomatis. In all of these studies, serovars were typed by comparing RFLP patterns of small fragments obtained by polyacrylamide gel electrophoresis.…”

TypingChlamydia trachomatis: from egg yolk to nanotechnology

“…The patterns were found to be compatible with the results obtained by serotyping. Rodriguez et al (1991) amplified 1200 bases of the omp1 gene and the product was digested by restriction nucleases, and resolved on polyacrylamide gels. Thirteen of the 15 serovars could be differentiated by this technique.…”

Comparison of reverse hybridization and ompA sequencing methods applied on Chlamydia trachomatis strains from Tunisia

“…This technique requires no specialized instruments and hence it was widely applied in epidemiological studies of C. trachomatis genotyping (Foschi et al, 2016;Petrovay, Németh, Balázs, & Balla, 2015;Rodriguez et al, 1991;Taheri Beni et al, 2012).…”