The Clinical Characteristic Of Chlamydia Trachomais Genotypes And Relationship With Cervical Intraepithelial Neoplasia

Chlamydia trachomais(CT) is a kind of specific microorganisms between virus and bacteria. It is an obligate intracellular bacterial pathogen because of the lacking or truncation of many biosynthetic pathways due to the small genome size (genome sizes of about 1 Mbp).Chlamydia trachomais is the leading cause of bacterial sexually transmitted infection worldwide, about 40% STD caused by CT, which makes it serious public health concern. CT normally infects the single cell columnar layer of epithelium of eye and urogenital tract. Due to the high percentage of asymptomatic patient, delayed or inappropriate treatment, persistent infections with long-term sequelae can develop mucopurulent cervicitis, pelvic inflammatory disease, infertility, ectopic pregnancy resulting from progressing and unrecoverable scarring. Recently, it is prompted that CT infection play role in cervix disease either favor the development of human papillomavirus (HPV)-induced neoplasia or direct impact.CT is traditionally classified into 15 main serovars based on the differential serospecificity of the major outer membrane protein (MOMP) or polymorphism of ompl (that encodes MOMP). CT present different tissue tropism that serovars A-C and Ba usually infect the ocular mucosa causing trachoma, the world’s leading cause of preventable infectious blindness, while serovars D-K are normally associated with noninvasive ano-urogenital infections, such as urethritis, epididymitis, prostatitis in males, cervicitis, urethritis, pelvic inflammation disease in females, and that serovars L1-3 are associated with invasive lymphogranuloma. Some animal experiment also showed that different serovars induced the variation in serological responses. But the associations between CT and risk of cervical intraepithelial neoplasia/cervical cancer(CIN/CC), CT genotype and clinical symptom remain controversial. With these considerations, the current study is proposed with following contests.At first, we performed a clinical observation for four years to investigate the association between CT infection and CIN with single infection or co-infection with HR-HPV. Then, we searched for studies in MEDLINE and VIP database to evaluate the risk of CT infection with CIN/CC used RevMan 5.2 software for Meta analysis.167 CT-positive genital swabs were analyzed by ompl gene PCR-RFLP and sequencing to study the distribution and clinical characteristics of CT genotypes.Part 1 Clinical observation of Chlamydia Trachomais infection and risk of the cervical intraepithelial neoplasiaObjective To reveal the association with CT infection and cervical intraepithelial neoplasia, or co-factor of human-papillomavirus(HR-PCR)Methods We perform a hospital-base cross-sectional study for 4 years. 12644 outpatients were involved in this study, Vaginal secretion samples of the patients were obtained to detect CT, HR-PCR and TCT,260 CIN patients were identified.Results The prevalence of CIN in patient group HR-HPV (+),CT (+), HR-HPV (+),CT (-), HR-HPV (-),CT (+), HR-HPV (-),CT (-) was 18.00%,10.26%,1.97% and 0.48%.Women with HR-HPV and/or CT infection had a significantly increased risk for CIN, especially at group of HR-HPV (+),CT (+).HR-HPV, CT infection, numbers of pregnancies, age were risk factors of CIN in the multiple Logistic regression model. Whether HR-HPV was positive or not, CT infection had positive association with CIN.Conclusion It is implied possible positive connection between CT infection and CIN. HR-HPV (+),CT (-), HR-HPV (-),CT (+), HR-HPV (-),CT (-) was 18.00%,10.26%,1.97% and 0.48%.Women with HR-HPV and/or CT infection had a significantly increased risk for CIN, especially at group of HR-HPV (+),CT (+).HR-HPV, CT infection, numbers of pregnancies, age were risk factors of CIN in the multiple Logistic regression model. Whether HR-HPV was positive or not, CT infection had positive association with CIN.Charpter 2 Chlamydia Trachomais infection and risk of Cervical intraepithelial neoplaisia/Cervical cancer: A meta analysisObjective To evaluate the risk of cervical intraepithelial neoplaisia/cervical cancer associated with Chlamydia Trachomais infection via document review and Meta analysis.Methods A systematic review and meta-analysis were conducted to summarize published literature on the association between CT and CIN/ICC. An extensive search of electronic databases MEDLINE(Pubmed), Cochrane Library, Embase, Web of Science and CBM was perform. The key words "Chlamydia trachomatis", "Chlamydia trachomatis infection" was used in combination with "low-grade squamous intraepithelial lesion", "high-grade squamous intraepithelial lesion", "cervical intraepithelial neoplasia" or "cervical cancer". Publications were included if they either reported odds ratios(OR) and corresponding 95% confidence intervals(CI) representing the magnitude of association between these two conditions, or presented data that allowed calculation of the OR.Results Out of 219 articles,8 cases-controls cohort studied and 4 cross-sectional studied were selected. After testing of heterogeneity and publication bias, meta-analysis was performed, using a random effects model. Although heterogeneity among studies was existed(Chi2=17.57, P<0.00001, I2=72%), a positive association between CT and CIN/ICC was found in cohort studied(OR 2.10,95%CI 1.68-2.63), in cross-sectional studied(OR 2.89,95%CI 2.15-3.90), respectively.Conclusion CT infection might be a co-factor of HR-HPV or an independent risk factor for CIN.Charpter 3 Chlamydia Trachomais genotyping methods with ompl PCR-RFLP and sequencingObject To build up the PCR reaetion system and form reference RFLP restriction maps by combinating CT ompl PCR-RFLP and ompl sequencing.Methods The cervical swabs of positive CT-NAAT were collected for serovar genotyping in Liuzhou people’s hospital from Jan,2010 to May,2014. We employed ompl gene PCR-RFLP combinate ompl gene sequencing, including DNA abstraction, selection of primers for PCR, establislishment of PCR reaetion system, ompl gene sequencing and BLAST, to form reference restriction enzyme maps undered our experiment condition.Results CT has been classified to 15 basic serotypes based on immunogenic epitope analysis of the MOMP which coded by ompl gene through monoclonal or polyclonal antibodies. Now PCR-RFLP based on the 1.1kbp ompl gene in lengty were used for clinical research, but different primer and experiment condition may led to different result. We established gene ompl PCR-RFLP restriction endonuclease maps to facilitate the genetyping of clinical samples. Meanwhile, we discover several variations in genome sequences through BLASTA and multiple sequence alignment.Conclusions We successfully amplified CT ompl gene, genotyping of cervical samples via PCR-RFLP and sequencing and found reference restriction enzyme maps of CT genotypes under our experiment condition. experiment condition may led to different result. We established gene ompl PCR-RFLP restriction endonuclease maps to facilitate the genetyping of clinical samples. Meanwhile, we discover several variations in genome sequences through BLASTA and multiple sequence alignment..Charpter 4 The clinical characteristics and distribution of CT genotype infection and risk of cervical intraepithelial neoplasiaObjective To investigate the distribution of the Chlamydia trachomatis(CT) genovar in cervical disease.Methods 128 women were included,41 with cervical intraepithelial neoplasia(CIN) and 87 were normal, DNA extracted from positve swabs was amplified by omp1 PCR-RFLP and ompl sequencing. Then describe the genotype distribution in terms of the age, clinical symptoms.Results 128 CT positive specimens were successfully genotyped by omp1 gene sequencing out of 167 samples (including CIN with CT positive in the clinical observation mentioned in the charpter 1). The most prevalent chlamydial genotype was D (n=38,29.69%), followed by E (n=28,21.88%), G (n=21,16.41%) and F(n=16,12.50%), genotype J、H、K remains comparatively rare. We did not find genotype I. The higher incidence of CT infection was detected in young subjects. But the distribution of CT genotype was not find association with age and clinical feature; Genotype G and F were more frequently concurrent infection of other bacteria infection. Genotype G was associated with mucopurulent cervicitis and CIN, while E was found more frequently concurrent infection of HR-HPV. However genotype D was most prevalent in our study, but it is a relatively low-risk type.Conclusion These results provide information on distribution of genital CT genetypes among clinical samples. Such data could have implications for the control and vaccine development of C. Trachomatis.