青海藏区B型沙眼衣原体的分离培养与鉴定

发布时间：2018-06-08 19:02

本文选题：沙眼衣原体 + 沙眼　；参考：《微生物学通报》2017年06期

【摘要】：【目的】对青海藏区沙眼患者标本进行沙眼衣原体分离培养与鉴定。【方法】分别采集患者的单眼结膜和结膜囊拭子标本至1 mL样本保护培养基中。取50μL样本采用离心法感染BGM细胞,37°C培养72 h,连续传代3次,相差显微镜观察衣原体包涵体。对临床样本和分离株分别进行主要外膜蛋白基因ompA序列分析。【结果】共采集了45例活动性沙眼患者的115份临床样本,其中54份样本为ompA PCR阳性,15份样本为沙眼衣原体培养阳性。ompA分析发现,青海藏区沙眼衣原体有3个不同的同源ompA变异株,均为基因B型,都包含有一个泌尿生殖道型沙眼衣原体特有密码子。分离株QH111L和QH111R分别来自编号111患者的左眼和右眼样本,它们ompA基因的可变区有一个非同义碱基差异。该碱基变异仅存在于111号患者的左眼样本中,说明QH111L可能是新出现的ompA突变体。【结论】青海藏区的眼型沙眼衣原体流行株为基因B型,至少存在3个不同的ompA变异株。从青海藏区分离培养了15株眼型沙眼衣原体,发现同一患者的左右眼样本中的沙眼衣原体有不同ompA。本研究为研制沙眼疫苗和诊断试剂奠定了基础,也将有助于理解沙眼的进化和传播。
[Abstract]:[objective] to isolate and identify Chlamydia trachomatis from Tibetan trachoma patients in Qinghai Province. [methods] Monocular conjunctiva and conjunctival sac swabs were collected into 1 mL culture medium. BGM cells were infected with 50 渭 L samples by centrifugation for 72 h and then cultured for 3 times. The inclusion bodies of chlamydia were observed by phase contrast microscope. The main outer membrane protein gene ompA sequences were analyzed in clinical samples and isolates. [results] A total of 115 clinical samples were collected from 45 active trachoma patients. Among them, 54 samples were ompA PCR positive and 15 samples were chlamydia trachomatis culture positive. OompA analysis showed that there were three homologous ompA variants of Chlamydia trachomatis in Tibetan area of Qinghai Province, all of which belonged to gene B type. Both contain a genitourinary Chlamydia trachomatis specific codon. The isolates QH111L and QH111R were collected from the left eye and right eye of patients with 111.The variable region of their ompA gene had a nonsynonymous base difference. QH111L may be a new ompA mutants. [conclusion] Chlamydia trachomatis in Qinghai Tibetan region is a gene B type, and there are at least three different ompA variants. Fifteen strains of chlamydia trachomatis were isolated from Qinghai Tibetan area. This study lays a foundation for the development of trachoma vaccine and diagnostic reagent, and will be helpful to understand the evolution and transmission of trachoma.
【作者单位】：首都医科大学附属北京同仁医院;病原微生物生物安全国家重点实验室军事医学科学院微生物流行病研究所;北京市眼科研究所;
【基金】：国家自然科学基金项目(No.31570177) 病原微生物生物安全国家重点实验室自主课题(No.SKLPBS1409)~~
【分类号】：R777.32

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