Rapid genotyping of the Chlamydia trachomatis major outer membrane protein by the polymerase chain reaction

Abstract

Twenty one Chlamydia trachomatis reference strains and 40 clinical isolates belonging to the lymphogranuloma venerum (LGV) and trachoma biovars were genotyped by differential restriction mapping of the major-outer-membrane-protein gene (MOMP) obtained by the polymerase-chain reaction (PCR). AluI digestion of the PCR product distinguishes eight MOMP-genotypes corresponding to 8 serovars. Six additional enzymes (NlaIII, CfoI, EcoRI, HinfI, DdeI and FokI) further permit the discrimination of 10 MOMP-genotypes corresponding to the 10 remaining serovars of the species. AluI alone allows direct typing of 78% of the clinical isolates. AluI digestion patterns of mouse C. trachomatis biovar, a C. pneumoniae and two C. psittaci strains, studied for comparison, were clearly distinguishable from one another and from the C. trachomatis LGV and trachoma strains. These results indicate that MOMP genotyping by PCR is a valuable molecular tool for studying C. trachomatis epidemiology.

Combination of PCR Targeting the VD2 of omp1 and Reverse Line Blot Analysis for Typing of Urogenital Chlamydia trachomatis Serovars in Cervical Scrape Specimens

“…In addition, because the RLB method uses a nonprecipitating ECL substrate, it has the advantage that nylon filters can be stripped and reused at least 10 times (16). Based on the fact that the VD2 primers are located within the omp1 gene, our RLB system has a further advantage in that it can also be used for typing of the PCR-amplified omp1 gene used for RFLP (4,8,12,13).…”

“…These techniques include standard MOMP serotyping, restriction fragment length polymorphism (RFLP) analysis of the PCRamplified omp1 gene, and nucleotide sequencing of the omp1 gene (3,4,12,13,19). Compared to immunotyping, the genotyping methods are more sensitive and specific for C. trachomatis serovar identification.…”

Predominance of Chlamydia trachomatis Serovars Associated with Urogenital Infections in Females in New Delhi, India

“…Epidemiological studies of C. trachomatis infections in sexual contacts have been few to date, which hampers the study of chlamydial transmission, its route of spread in a population, its virulence factors, and the associated risk factors of C. trachomatis infections (12,19). Compared with immunotyping, the genotyping methods, particularly omp1 are more sensitive and precise in revealing C. trachomatis variants within serovars as well as in potential recombinants among serovars (13,15,18). The present study was undertaken to understand the occurrence of C. trachomatis serovars in the genital tracts of infected women, which will help in devising an effective screening program for routine diagnosis of chlamydial infections, understanding the immunopathogenesis, and developing an effective chlamydial control program.…”

Restriction endonuclease patterns of the omp1 gene of reference Chlamydia trachomatis strains and characterization of isolates from Cameroonian students

“…Serovars A-C are most often associated with trachoma and serovars L1-L3 are associated with lymphogranuloma venereum, while serovars D-K are primarily associated with urogenital and neonatarum infections (Morré et al, 1998a;Ossewaarde et al, 1994;Sturm-Ramirez et al, 2000). Various studies have shown the feasibility of typing clinical isolates by PCR-based RFLP analysis of the amplified omp1 gene encoding MOMP (Frost et al, 1991;Rodriguez et al, 1991;Sayada et al, 1991), avoiding the costly production of monoclonal antibodies. This method was originally performed on cell cultures of cervical and urethral specimens.…”