Genotyping of Portuguese Chlamydia trachomatis urogenital isolates.

OBJECTIVE: To determine the prevalence of the different Chlamydia trachomatis genotypes in Portuguese patients. METHODS: Urogenital isolates (n = 240) derived from attenders of various clinics in the Lisbon area were differentiated into genovars by genotyping with restriction fragment length polymorphism (RFLP) analysis of the PCR amplified omp1 gene. RESULTS: Genotype E was the most common for both men (47.9%) and women (43.8%). Genotypes D and F were the second most prevalent for men (11.3%) and genotype H was the second most prevalent for women (19.5%). Genotypes F, G, D, in women and H, G, I, in men, were found in a lower percentage of cases. Genotypes B, Ba, J, K, L1 and L2 were very rarely detected. CONCLUSIONS: With one exception, the overall distribution of Chlamydia trachomatis genotypes in our study is similar to what has been observed in other western countries. The only exception is the unusual prevalence of genotype H among women. The clinical manifestations associated with this and other genotypes were similar.     

沙眼衣原体的基因分型及其临床意义

目的　探讨对沙眼衣原体更为简便的分型方法及基因分型的临床意义。方法　利用聚合酶链反应 (PCR)扩增沙眼衣原体外膜蛋白基因 (omp1 )片段 ,以核酸内切酶作酶切 ,1 0 %聚丙烯酰胺凝胶电泳 ,银染色后观察带型分布。结果　扩增 1 5种血清型omp1基因 ,产物为 871bp的DNA片段。用AluⅠ酶切此片段 ,观察到 1 5种血清型呈现具有特征性的带型分布 ,但C组带型较难区别。再用HpaⅡ、HinfⅠ及EcoRⅠ三重酶切 ,则可将C组各型完全区分开来。对 74株沙眼衣原体临床株进行分型 ,发现E ,F ,G ,D在本观察人群中占优势 ,偶尔 ,也见到B ,H ,J等型别。还观察到 1例F/D混合型。沙眼衣原体的型别与临床表现之间未见有明显的关联 ,但发现D型感染者常为抗沙眼衣原体抗体高滴度者。结论　omp1基因分型技术可以作为流行病学调查中的有用的研究工具     

Genotyping of Chlamydia trachomatis from endocervical specimens in Brazil

BACKGROUND: There is no data concerning genotyping of Chlamydia trachomatis from Brazilian samples. GOAL: To characterize the genotype of C. trachomatis detected in women assisted at a STD public clinic and establish the prevalence of this infection in that population. STUDY DESIGN: Endocervical samples of a group of 100 women were tested for chlamydial infection with PCR directed to C. trachomatis cryptic plasmid. Genotyping of positive samples were done after omp1 amplification and sequencing. RESULTS: The overall prevalence of C. trachomatis infection was 19%, with the highest prevalence in women between 15 and 25 years old (68.4%). Four genotypes were found associated with endocervical infections: D, E, F, and K. Sequence analysis revealed a coinfection of genotypes D and E in 1 woman. CONCLUSIONS: To our knowledge this is the first study to characterize Brazilian C. trachomatis endocervical samples and Brazilian C. trachomatis genotype coinfection. Our results also emphasize the importance of routine diagnosis of C. trachomatis for the control of this STD.     

多重聚合酶链反应用于D－K型沙眼衣原体基因分型的研究

目的摸索一套泌尿生殖道沙眼衣原体基因分型的多重聚合酶链反应法。方法选择某一型与其它各型均不同的可变区部位作为下游引物,以恒定区 1区的高度保守区作为各型的公用上游引物,优化反应条件,进行标准株和临床野生株的多重聚合酶链反应扩增,并与聚合酶链反应－限制性内切酶片段多态性方法比较。结果用摸索出的反应系统和条件,对 8个标准株进行扩增,扩增出与理论值一致的片段,并可直接在普通水平琼脂糖凝胶进行电泳。特异性检测证明：①衣原体各型之间没有非特异性交叉扩增,②对泌尿生殖道病原和寄生微生物进行扩增,未扩出任何 DNA带。对临床野生株分型结果证明：复合聚合酶链反应阳性率为 100％,而聚合酶链反应－限制性内切酶片段多态性为 94.44％。结论本套衣原体基因分型的多重聚合酶链反应法特异、敏感、快速、实用,最终能用于临床分型。     

Chlamydia trachomatis

<正>0001 Chlamydia trachomatis utilizes the mammalian CLA1 lipid transporter to acquire host phosphatidylcholine essential for growth沙眼衣原体利用哺乳动物CLA1脂质转运获取生长必须的宿主磷脂酰胆碱     

性病门诊男性尿道炎患者沙眼衣原体基因分型

目的 了解性病门诊就诊的尿道炎男性患者中,沙眼衣原体血清型分布情况。 方法 采集2013年1 - 12月中国医学科学院皮肤病医院性病门诊有尿道炎症状的男性患者的尿液,荧光定量PCR检测沙眼衣原体,对沙眼衣原体阳性患者的尿液提取DNA,用巢式 PCR法扩增沙眼衣原体主要外膜蛋白基因ompA的VS1-VS2片段,然后对此片段测序,测序结果用DNAStar5.0软件与每种血清型的参考菌株做比对,分析其血清型。 结果 对2013年432例男性尿道炎患者进行了沙眼衣原体筛查,阳性标本143例,阳性率33.1%。143例沙眼衣原体阳性标本,127例扩增出ompA的VS1-VS2片段,16例未扩增出。127例阳性标本经测序分析获得9种血清型。血清型分布情况如下：E型29（22.83%）、F型28株（22.05%）、D型19（14.96%）、G型16株（12.60%）、J型16株（12.60%）、K型8株（6.30%）、H株5株（3.94%）、I型3株（2.36%）、B型3株（2.36%）,E、F、D、J、G型占85.02%。与标准菌种比对,发现127例菌株中有14株存在碱基突变,为同义突变。 结论 性病门诊男性尿道炎沙眼衣原体血清型主要是E型、F型、D型和G型,与20年前相比,E型菌株比例有所下降,J型菌株比例增高。     

南京地区性病患者沙眼衣原体感染的基因分型

目的了解南京地区性病患者泌尿生殖道沙眼衣原体感染分离株的基因型（血清型）分布情况。方法应用聚合酶链反应和限制性片段长度多态性分析,对来自南京地区性病门诊病人和性乱妇女的９４份沙眼衣原体阳性标本进行了基因分型研究。结果Ｅ型为南京地区性病高危人群最流行的沙眼衣原体基因型,占３６．２％,其次为Ｆ、Ｇ和Ｄ型,分别占１９．２％、１７．０％和１２．８％,而Ｋ、Ｊ、Ｉ、Ｈ型比较少见。发现了一个Ｂ基因型,一个Ｆ／Ｇ混合型和４个非典型基因型。结论我国南京泌尿生殖道沙眼衣原体感染流行株的基因型分布情况与国外基本相同,且存在某些基因型的变异。     

Chlamydia trachomatis diagnostics

Nucleic acid amplification (NAA) assays for the diagnosis of Chlamydia trachomatis infections started to appear in the peer reviewed literature about 12 years ago and during that period we have seen an incredible effort put into the development and evaluation of commercially developed NAA kits to diagnose and treat infections.     

沙眼衣原体omp1基因VS1-VS2序列分析与基因分型

目的 探讨沙眼衣原体omp1基因VS1-VS2序列基因分型和VS1-VS2基因突变率。方法 巢式PCR扩增omp1基因的VS1-VS2序列,自动测序仪测序,运用DNAstar软件进行临床菌株与标准菌株序列的比对。结果 99株临床标本分离株扩增出约453 bp大小的VS1-VS2片段。与标准株比对,除3株测序出现双峰无法确定基因型外,96株菌株分为9个型,以F型29株(30.2%)、E型22株(22.9%)、J型19株(19.8%)、D型12侏(12.5%)和H型8株(8.3%)为主。序列分析发现7株存在VS1区碱基的突变或插入,多为有义突变,突变率为7.3%,多见于D、E、F和H型。结论 沙眼衣原体omp1基因序列分析在分子流行病学上具有一定意义。     

Chlamydia trachomatis genital infections

The prevalence and clinical features of Chlamydia trachomatis infection were studied in men with nongonococcal urethritis (NGU), in their female sexual partners, and in other women. Of 92 patients with NGU, 36 (39%) were chlamydia-positive; although all had symptoms of urethritis, fewer than half had evident discharge. Clinical features of chlamydia-positive and chlamydia-negative NGU were indistinguishable. Most female contacts of chlamydia-positive patients with NGU were infected with C. trachomatis; contacts of chlamydia-negative patients with NGU seldom harboured chlamydia. Inflammatory and erosive changes were equally common in the uterine cervix of chlamydia-positive and chlamydia-negative women, although almost half of infected cases had no clinical signs of infection and no specific symptoms. In such women the present unavailability of sensitive screening techniques is a major obstacle to the effective control of chlamydial infection.     

中国南方部分城市泌尿生殖道沙眼衣原体基因分型研究

目的 了解中国南方4个城市性病高危人群中沙眼衣原体(Ct)基因型的分布特征。方法 从衡阳、上海、广州、江门四个城市的性病门诊收集1180份泌尿生殖道标本,经质粒PCR筛选后,阳性者用巢式PCR扩增Ctomp1基因片段,酶切omp1扩增产物,分析Ct各临床株omp1基因的限制性片段长度多态性(RFLP),将临床株分型。结果 Ct质粒PCR阳性者301份,用于分型者286份。通过RFLP分型,共检出10个基因型,其中常见的是E(30.8%)、J(23.8%)、D(17.5%)、F(10.5%)、G(6.2%)型,并且不同城市间基因型的分布差异无统计学意义(χ²=0.981,P>0.05)。结论 4个城市存在Ct多种基因型,并且基因型的分布大致相同。