沙眼衣原体参考菌株omp1基因的限制性内切酶图谱及喀麦隆学生分离株的特征分析

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

作者信息

Ngandjio Antoinette, Clerc Maithe, Fonkoua Marie Christine, Thonnon Jocelyn, Lunel Françoise, Bébéar Christiane, Bianchi Anne, de Barbeyrac Bertille

机构信息

Centre Pasteur du Cameroun, Yaounde, Cameroon 2Laboratoire de Bactériologie, Centre National de Référence des infections à Chlamydia, Université de Bordeaux II, Zone Nord, Bât. 2B, 146 Rue Léo Saignat, 33076 Bordeaux, France 3Institut Alfred Fournier, Paris, France 4Laboratoire Départemental, Conseil Général de Seine Saint Denis, France.

出版信息

J Med Microbiol. 2004 Jan;53(Pt 1):47-50. doi: 10.1099/jmm.0.05333-0.

DOI:10.1099/jmm.0.05333-0

PMID:14663104

Abstract

Eighteen reference strains of Chlamydia trachomatis were differentiated by omp1 PCR- and nested PCR-based RFLP analysis, using two restriction digestions, one with AluI and the other with the three enzymes HpaII, EcoRI and HinfI. AluI digestion allowed the differentiation of 12 different profiles after CT1/CT5 PCR and 13 different profiles after the nested PCR. The triple hydrolysis permitted the identification of 15 different patterns. In all, 16/18 reference strains were clearly identified. These reference patterns were successfully used to genotype 34 of 35 (28 strains and 7 clinical specimens) samples from infected students, collected during a screening programme in Yaounde (Cameroon). Genotypes D, Da, E, F, G and J were found. The most prevalent omp1 genotype was E (n = 14; 40 %), followed by F (n = 7; 20 %). As RFLP patterns of reference strains are essential for typing clinical isolates, they will greatly facilitate C. trachomatis characterization in many resource-limited laboratories.

摘要

采用omp1聚合酶链反应（PCR）和基于巢式PCR的限制性片段长度多态性（RFLP）分析，使用两种限制性酶切方法（一种用AluI，另一种用HpaII、EcoRI和HinfI这三种酶）对18株沙眼衣原体参考菌株进行区分。AluI酶切在CT1/CT5 PCR后可区分出12种不同的图谱，在巢式PCR后可区分出13种不同的图谱。三重酶切可鉴定出15种不同的模式。总共16/18株参考菌株被明确鉴定。这些参考模式成功用于对来自喀麦隆雅温得一项筛查项目中感染学生的35份样本（28株菌株和7份临床标本）中的34份进行基因分型。发现了基因型D、Da、E、F、G和J。最常见的omp1基因型是E（n = 14；40%），其次是F（n = 7；20%）。由于参考菌株的RFLP模式对于临床分离株的分型至关重要，它们将极大地促进许多资源有限实验室对沙眼衣原体的鉴定。