Distinguishing Chlamydia species by restriction analysis of the major outer membrane protein gene

Clinical isolates of Chlamydia pneumoniae from diverse geographic locations and strains of other Chlamydia species were typed by polymerase chain reaction (PCR) amplification of the major outer membrane protein (MOMP) gene followed by restriction fragment length polymorphism analysis of the product. Use of synthetic primers corresponding to highly conserved regions of the MOMP gene resulted in amplification of a 1070 bp product in laboratory strains and clinical isolates of C. pneumoniae, C. trachomatis and C. psittaci. PCR products were digested with restriction enzymes Alu I and Mbo I and separated by polyacrylamide gel electrophoresis. Restriction fragment patterns varied in length from 8-12 bands of 30-400 bp in size in Alu I digests, and 6-7 bands of 50-400 bp in size in Mbo I digests. Strains representing different chlamydia species were easily distinguishable by this method, as were different serovars of C. trachomatis. Strains of C. pneumoniae tested include laboratory strain TW-183 and recent clinical isolates from Atlanta, Brooklyn, Wisconsin and Norway. One combination of primers reacted with C. psittaci strains and C. pneumoniae strain TW-183, but not with other strains of C. pneumoniae tested regardless of the concentration of DNA in the sample. With use of a pan-reactive primer combination, however, restriction patterns were similar in all strains of C. pneumoniae tested. This gene typing technique can be valuable for distinguishing the three chlamydial species and potentially strains of C. pneumoniae in clinical and epidemiologic studies.