Rodriguez P, Vekris A, de Barbeyrac B, Dutilh B, Bonnet J, Bebear C
Laboratoire de Bactériologie, Université de Bordeaux II, France.
J Clin Microbiol. 1991 Jun;29(6):1132-6. doi: 10.1128/jcm.29.6.1132-1136.1991.
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 L1 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 L1. Computer analysis of these sequences indicated that two restriction digestions performed in parallel, one with AluI and the other with IIpaII, followed by HinfI and EcoRI, would allow the theoretical differentiation of 13 serovars. Serovars Ba and L1 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.
开发了一种通过对主要外膜蛋白(MOMP)的扩增基因进行限制性内切酶分析来鉴定沙眼衣原体菌株的方法。对15个血清型(A至K以及L1至L3)的参考菌株和临床分离株进行了检测。利用每个血清型已知的四个可变区序列和血清型L1的恒定区序列,任意构建了15个血清型MOMP基因的核苷酸序列。对这些序列的计算机分析表明,并行进行两次限制性消化,一次用AluI,另一次用IIpaII,然后用HinfI和EcoRI,将能够从理论上区分13个血清型。血清型Ba和L1呈现相同的理论限制性图谱。我们的分型方法包括对MOMP基因约1200 bp的片段进行聚合酶链反应扩增,然后用上述酶进行限制性内切酶消化。从15个血清型中,我们获得了14种不同的图谱;13种图谱是血清型特异性的,而血清型B和Ba呈现相同的图谱。将这种分型方法应用于从临床材料中分离的沙眼衣原体菌株,对于17株中的14株,其结果与免疫分型方法相同。此外,限制性内切酶分析检测到了一个血清型内的差异。这种方法对于流行病学研究似乎很有前景。