福州市性病门诊人群生殖道沙眼衣原体的分子流行病学研究

目的 探讨福州市性病门诊人群泌尿生殖道沙眼衣原体的基因型分布状况。方法 收集2013-2014年临床疑似生殖道沙眼衣原体感染者标本2 019份,用实时荧光定量PCR检测沙眼衣原体,阳性标本用套式PCR扩增Omp1区并进行测序分析。测序结果上传至BLAST网站查找序列的相似性、构建系统树确定基因型。结果 检测沙眼衣原体阳性标本86份,成功分型的有83份,1株为混合感染。测得8个基因型,分别为:F型(36.59%)、E型(24.39%)、D型(12.20%)、G型(9.76%)、J型(9.76%)、H型(3.66%)、K型(2.44%)、B型(1.22%)。计算不同基因型间Omp1基因的核苷酸的同义突变率(d_S)和非同义突变率(d_N),发现仅G型的d_N/d_S值大于1,其他型均小于1。结论 F型和E型是福州市性病门诊人群泌尿生殖道沙眼衣原体的主要基因型。同义替代是Omp1基因在进化过程中的主要变异。     

湘西地区女性泌尿生殖道沙眼衣原体感染状况及基因分型初步研究

目的对湘西地区女性泌尿生殖道沙眼衣原体的感染状况及基因分型进行初步研究。方法利用单克隆抗体直接免疫荧光法（DFA）对100例宫颈病变患者及77位健康女性进行筛查,对沙眼衣原体感染者的宫颈管内拭子标本直接进行套式PCR,扩增主要外膜蛋白基因（Omp1）第4可变区（VS4）,对将扩增的Omp1VS4基因片段进行测序、分型,并进行同源性分析。结果DFA检测177名妇女的宫颈拭子标本,36份沙眼衣原体阳性,29份阳性标本扩增出277bp目的DNA。经基因测序,共检出8个基因型,其中E型9株（31.0%）、F型7株（24.1%）、G型5株（17.2%）、D型3株（10.3%）、H型2株（6.9%）、J型1株（3.5%）、Ba型1株（3.5%）、K型1株（3.5%）,4例存在Omp1基因变异。结论湘西地区女性存在生殖道沙眼衣原体感染,利用对基因的多态性分析可以对沙眼衣原体感染流行情况作出分析。     

性传播疾病门诊2268例就诊者病原检测分析

目的了解性传播疾病（性病）门诊就诊者泌尿生殖系统病原体感染情况。方法对2005年6月-2007年5月在淮安市第四人民医院性病门诊就诊的2268例患者泌尿生殖道病原体进行检测。结果2268例患者中，有366例检出病原体，阳性率16．14％（366／2268）。单一病原体感染率为69．13％（253／366），其中检出淋球菌68份（42．5％），沙眼衣原体80份（44．44％），解脲支原体153份（68．91％），梅毒23份（27．7％），生殖器单纯疱疹9份（52．94％），尖锐湿疣27份（24．55％）：混合感染为30．87％（113／366），其中检出沙眼衣原体＋解脲支原体＋淋球菌42份（12．88％），沙眼衣原体＋解脲支原体＋梅毒螺旋体12份（6．25％），HPVF＋HIV1份（1．67％），HPV＋梅毒螺旋体2份（3.77％）。结论门诊就诊者中，中青年患者居多，应重视病原学检查，强化性病防治力度。     

下呼吸道感染患儿沙眼衣原体检测与初步基因分型

研究下呼吸道感染患儿沙眼衣原体（Ct）的检测情况、Ct分离株的初步基因分型及方法学评价。采用McCoy细胞培养、聚合酶链反应（PCR）对下呼吸道感染患儿248例鼻咽拭子（其中110例加下睑结刮取标本）进行检测；8份患儿Ct标本经双链DNA直接测序法以及裂解酶片段长度多态性（CFLP）基因分型，下呼吸道感染患儿鼻咽标本Ct检出率为25％（62／248），其中110例新生儿鼻咽和结膜标本Ct总检出率为40.9%（45／110），基因分型：E型4株，F、D和H型分别为2、1和1株，6株成人性病株中E型3株，F、D和H型各1株。取E型3份，F型2份作CFLP，呈两类CFLP谱，提示同时取患儿鼻咽和结膜标本可提高Ct检出率，下呼吸道感染婴儿的Ct型别与成人性病Ct型别关系密切，双链DNA直接测序法及CFLP均可用于Ct基因分型。     

11254例生殖道沙眼衣原体感染荧光PCR检测结果分析

目的对宝安区人民医院门诊病人生殖道沙眼衣原体感染,经荧光聚合酶链式反应（PCR）检测结果进行分析,探讨其感染现状及临床检测意义。方法采用荧光PCR方法检测11254例患者的沙眼衣原体（CT）的脱氧核糖核酸（DNA）．结果11254例患者标本中共检出832例阳性,总阳性率为7．39％;其中男性检测3612例,阳性数426例,女性7640例,阳性数406例,阳性率分别为11．79％,5．31％;男性女性发病比率差异有统计学意义（x^2=150．167,P〈0．001）。沙眼衣原体阳性患者高发年龄为20～39岁,占84．86％;支原体的合并感染率为14．18％。结论临床医生应注意临床病人多重感染的情况,应对全社会进行积极的健康的性教育与宣传,普及检测手段。     

1996～2003年上海市某医院泌尿生殖道分泌物标本性病检测结果分析

目的了解上海市近几年性病门诊就诊者中淋球菌(NG)、沙眼衣原体(CT)和解脲脲原体(UU)的流行趋势和特征，探讨可能的影响因素，为制定和开展有效的防治措施提供参考。方法收集1996～2003年到上海市皮肤病性病医院性病门诊就诊，并进行泌尿生殖道分泌物标本性病检测者的人口学特征和检测结果等资料，进行统计学描述和分析。结果在此期间共有66658人接受检查，其中NG阳性18348例(35．8％)，CT阳性7048例(18．1％)，NG和CT阳性人数和阳性率在1997年和1998年达高峰，之后下降，近年又开始上升。UU阳性6673例(37．4％)，其阳性人数和阳性率则基本上呈逐年上升趋势。混合阳性率最高的是CT和UU(14．2％)，最低的是NG和UU(8。7％)。女性NG阳性者中的CT阳性率(35．7％)和CT阳性者中的NG阳性率(27．2％)，分别高于所有受检者对应的阳性率(20．4％和19．6％)，男性情况相反。结论上海地区性病流行谱正在发生改变，女性UU感染人数明显增多；近年来NG、CT和UU的混合感染率处于较高水平，尤其是低龄女性；NG和CT的混合感染情况应该引起重视。     

郑州地区936例泌尿生殖道感染者支原体培养结果分析

对本站性病门诊泌尿生殖道患者９３６例标本进行支原体培养及沙眼衣原体检测,结果支原体培养阳性２１５例（２３．０％）,ＵＵ感染为１７６例（１８．８％）显著高于ＭＨ感染１０例（１．１％）和ＵＵ＋ＭＨ混合感染２９例（３．１％）（Ｐ＜０．０１）。在２１５例支原体阳性患者中合并Ｃｔ感染者４２例（１９．５％）,另对２１５例患者不同性别、年龄的支原体感染、合并感染进行了分析。     

云南省某市女性性工作者HIV感染及其危险因素调查

目的 了解云南省某市女性性工作者（FSWs）艾滋病病毒（HIV）和性传播疾病（盯D）韵感染情况及HIV感染相关危险因素。方法 招募FSWs调查其一般人口学及行为学资料，采集静脉血进行HIV、单纯疱疹病毒2型（HSV-2）和梅毒血清学检测，采集尿液进行尿吗啡检测，采集宫颈分泌物进行淋球菌和沙眼衣原体检测，采集阴道分泌物进行阴道毛滴虫检测。结果 共收集血液标本737份，其中76份经确认为HIV抗体阳性，感染率为10．3％，梅毒感染率为7．5％，HSV-2感染率为68．1％。共收集阴道和宫颈分泌物标本各737份，淋病感染率为8．3％，沙眼衣原体感染率为25．9％，阴道毛滴虫感染率为10．6％。共收集尿标本739份，吗啡阳性率为15．6％。多因素logistic回归模型分析显示，与HIV感染有关的危险因素是：低档性服务场所服务，静脉注射吸毒，过去一年出现下腹部疼痛，生殖器溃疡，外阴增生物，从事性服务时间≥5年，HSV-2阳性。结论 当地FSWs的HIV和STD感染率较高，需针对该人群采取有效的干预措施，及时治疗STD，控制HIV的传播流行。     

泌尿生殖道标本中沙眼衣原体的直接检测及基因分型

应用聚合酶链反应（PCR）和限制性片段长度多态性（RFLP）分析，建立了沙眼衣原体（Ct）直接检测及基因分型的方法。与细胞培养法比较，评价其检测的敏感性。400份性病门诊患者的泌尿生殖道标本同时用培养和质粒PCR检测Ct，结果29份标本培养阳性，其中28份标本质粒PCR阳性，另有5份培养阴性的标本质粒PCR阳性。对培养和质粒PCR阳性的标本进一步做主要外膜蛋白基因（ompl）PCR或ompl巢式PCR，结果32份标本omplPCR或ompl巢式PCR阳性。将阳性标本的ompl产物用AluI和MspI等限制性内切酶酶切，产生的酶切带谱与标准株带谱比较，以鉴定阳性标本的基因型。结果E型为13份标本，F：6、G：4、D：3、J：2、K：2、H：1、非典型：1。研究表明：PCR－RFLP可直接检测及基因分型泌尿生殖道标本中的Ct，为一种敏感的血清分型替代方法。     

福州市生殖道沙眼衣原体的MLST分子流行病学研究

目的 研究多位点序列分型法(MLST)在福州地区性病门诊人群生殖道沙眼衣原体基因分型中的分辨力,并与来自阿姆斯特丹性病门诊女性患者的样本进行比较。方法 采用MLST分型法研究福州性病门诊生殖道沙眼衣原体感染者的基因型,eBURST进行ST型地理分布特征分析。结果 从86份阳性标本中可获得完整MLST数据的样本为76份,共分30个ST型,其中14个为新的型别。MLST分型法的Simpson分辨力指数为0.9。eBURST分型结果显示30个ST型分为3个克隆复合体和5个独特型。仅有4个ST型在福州和阿姆斯特丹两地同时存在。结论 MLST为生殖道沙眼衣原体分子流行病学调查提供高分辨率的分型手段。eBURST 分析可揭示不同地理来源的生殖道沙眼衣原体之间的进化关系。     

Genotype distribution of genital Chlamydia trachomatis in Chiang Mai,Thailand

Urogenital isolates (N=84) of Chlamydia trachomatis collected from high-risk STD subjects in Chiang Mai and the surrounding areas were investigated for genotype distribution. C. trachomatis genotypes were determined by the PCR-based RFLP technique and confirmed by nucleotide sequencing. By this method, the VD4 DNA of the MOMP gene was amplified and digested separately with 4 restriction endonucleases, AluI, HindIII, DdeI, and EcoRII. The nucleotide sequence was determined by dye terminator cycle sequencing. Eight different C. trachomatis genotypes were identified: genotype D (34.5%), F (21.4%), K (13.1%), H/Ia (8.3%), E (7.1%), B/Ba (7.1%), G (6.0%), and J (2.4%). Genotype D and F were the commonest, accounting for 56% of the C. trachomatis infections. When nucleotides of the VD4-MOMP gene were anlyzed, 43 samples (51.2%) had nucleotide sequences that differed from the prototypes, while 41 (48.8%) were identical. Nucleotide substitution mutation was the major mechanism in these variants; changes in nucleotide sequences usually resulted in amino acid substitution, which could lead to a modification of antigenic determinants and the consequent evasion of immune responses.     

Evaluation of diagnostic efficacy of PCR methods for Chlamydia trachomatis infection in genital and urine specimens of symptomatic men and women in India

In India, given the scarce availability of sensitive and specific methods, Chlamydia trachomatis genital infections may lead to severe clinical complications when left undiagnosed or underdiagnosed. The present study was conducted to evaluate the diagnostic efficiency and feasibility of polymerase chain reaction (PCR) assays using genital and urine specimens from men and women in India. Genital swabs and urine specimens collected from 143 patients attending the sexually transmitted disease (STD) clinic, Government General Hospital, Chennai, were tested by culture and a plasmid based PCR. Culture was positive in 27 (18.9%) patients. PCR gave positive results for 46 (32.2%) cases using genital specimens, and the positivity rate in urine was 25.2%. Once the discordant results between culture and PCR had been resolved by using a major outer membrane protein PCR, the overall sensitivity, specificity, and positive and negative predictive values for the plasmid PCR in genital specimens were 100%, 98%, 95.7%, and 100%, respectively. Corresponding values for urine PCR were 81.8%, 100%, 100%, and 92.5%, respectively. The prevalence of confirmed C. trachomatis infection was 30.8% in this STD population. The results confirmed the need to use sensitive and specific molecular assays like PCR to prevent underdiagnosis of genital chlamydial infections and to facilitate better clinical management of this infection in India.     

Coinfection of Chlamydia trachomatis, Ureaplasma urealyticum and human papillomavirus among patients attending STD clinics in Estonia

Women visiting Estonian STD clinics were subjected to PCR assay for human papillomavirus (HPV), Chlamydia trachomatis and Ureaplasma urealyticum biovar 2. The overall prevalence of coinfection was 8%. The chlamydial infection was found to be associated with HPV, especially with high-risk HPV (OR=2.5, p<0.005) and most significantly in women over 41 y of age. C. trachomatis infection also occurred more frequently in U. urealyticum-infected than in U. urealyticum-free patients (OR=2.6, p=0.02). U. urealyticum infection did not associate with HPV status. The clinical significance of the association between C. trachomatis and U. urelyticum infection remains to be elucidated.     

OmpA-VS2为靶基因的非标记实时荧光PCR检测沙眼衣原体感染

目的建立一种以OmpA-VS2为靶基因的非标记实时荧光聚合酶链反应(PCR)方法,以检测沙眼衣原体。方法设计15种型别沙眼衣原体通用引物,对OmpA-VS2进行套式PCR扩增和实时荧光PCR检测,并对其敏感性、特异性进行评价,并用临床标本进行验证。结果15种型别沙眼衣原体均扩增出168bp的目的片段;敏感性为每个反应1个拷贝。特异性分析可见,与10种泌尿生殖道病原体和共生菌均没有交叉反应。临床标本验证的结果显示,该方法与以质粒为靶基因的核酸扩增检测的Amplicor CT/NG方法的检测结果一致。结论该方法具有敏感性高、特异性强、扩增后无需PCR后处理等特点,可望用于临床与防治项目中沙眼衣原体的检测。     

Distribution of Chlamydia trachomatis serovars among female prostitutes and non-prostitutes in Thailand, and non-prostitutes in Japan during the mid-90s

The distribution of Chlamydia trachomatis serovars in Thailand and Japan during the same period of the mid-90s was determined. Seventy-one C . trachomatis specimens isolated from female patients who visited the Venereal Diseases Center at Bangkok, Thailand in 1994 were used in this study. Of these, 56 patients were prostitutes. Forty-seven specimens obtained from female non-prostitutes who attended the Department of Obstetrics and Gynecology, Saitama Medical School, Japan during the period from 1993 to 1995 were also used in this study. DNA was extracted from these specimens and typing of C. trachomatis serovars was performed by the polymerase chain reaction-restriction fragment length polymorphism method. The identified serovars among prostitutes of Thailand (n = 56)/non-prostitutes of Thailand (n = 15)/non-prostitutes of Japan (n = 47) were as follows: Ba 1/0/2, D 8/1/15, E 11/2/8, F 16/9/8, G 4/0/7, H 3/2/3, I 1/0/1, J 3/0/0, and K 10/1/4. Serovar F was most prevalent (35.2%) in both prostitutes and non-prostitutes from Thailand, followed by serovar E (18.3%). On the other hand, serovar D was the most frequent serovar in non-prostitutes in Japan (31.9%) followed by serovars F (17.0%) and E (17.0%). A difference in the distribution of C. trachomatis serovars of Thailand and Japan was noted.     

Simultaneous detection of Neisseria gonorrhoeae and Chlamydia trachomatis by PCR in genitourinary specimens from men and women attending an STD clinic

Neisseria gonorrhoeae and Chlamydia trachomatis are the two most common bacterial sexually transmitted infections that manifest primarily as urethritis in males and endocervicitis in females, though the infection may be asymptomatic especially in women. Since complications may occur in untreated symptomatic and asymptomatic infected individuals, early diagnosis and treatment of infected individuals is required to prevent severe sequelae and spread of these diseases. Recently molecular amplification assays like Polymerase Chain Reaction (PCR) and Ligase Chain Reaction (LCR) have been found to be highly sensitive and specific methods for detection of N. gonorrhoeae and C. trachonmatis not only in urethral and cervical specimens but also in urine. The objective of this study was to screen male and female Sexually Transmitted Disease (STD) clinic attenders, with and without symptoms suggestive of urethritis and cervicitis for presence of N. gonorrhoeae and C. trachomatis using a multiplex PCR based assay, to compare its performance with culture for N. gonorrhoeae and Direct Fluorescent Antibody (DFA) staining for C. trachomatis and also to compare the efficacy of PCR test performed on urine and genital swab specimens collected from this high risk group. Genital specimens and urine was collected from STD clinic attenders. N. gonorrhoeae and C. trachomatis was detected in genital specimens by culture and DFA respectively. Multiplex PCR was used to detect N. gonorrhoeae and C. trachomatis infection in both genital and urine specimens. Among men with urethritis, N. gonorrhoeae was detected in 70% by culture and 77% by PCR, while C. trachomatis as detected in 7.5% by DFA and 17.5% by PCR. Among females with endocervicitis, N. gonorrhoeae was detected in 7.7% by culture and 30.7% by PCR, while C. trachomatis was detected in 7.7% by DFA and in 15.4% by PCR. None of the asymptomatic males were positive for N. gonorrhoeae and C. trachomatis by conventional methods, while 43.9% were positive for N. gonorrhoeae and 7.5% for C. trachomatis by PCR. Fifty per cent of asymptomatic women were positive for C. trachomatis by PCR alone. We encountered PCR positive but culture/DFA negative results and also PCR negative but culture/DFA positive results. In view of this a single PCR test cannot be used for diagnosis and treatment of N. gonorrhoeae and C. trachomatis infection unless confirmed by a second test.     

热启动多聚酶链反应快速检测泌尿生殖道沙眼衣原体的应用研究

应用热启动多聚酶链反应技术,对泌尿生殖道沙眼衣原体感染进行快速检测。共检测８０６例泌尿生殖道炎症病人,其中ＳＴＤ病人４０４例,阳性１０５例,阳性率２６．０％;普通病人４０２例,阳性１０例,阳性率２．５％。经统计学估计,福州市ＳＴＤ患者沙眼衣原体感染率范围为２１．７～３０．３％。     

Usefulness of real-time PCR in detecting Chlamydia trachomatis and Neisseria gonorrhoeae in endocervical swabs and first-voided urine specimens

We evaluated performance of Abbott RealTime CT/NG assay (real-time PCR, Abbott Japan) for detect Chlamydia trachomatis and Neisseria gonorrhoeae by real-time PCR in 88 female patients with cervicitis symptoms seen at gynecological clinics and 100 male patients with urethritis symptoms seen at urological or dermatology clinics in Kitakyushu, Japan. Endocervical swab and first-voided urine (FVU) specimens were then collected from women and FVU specimens from men. Detection rates of C. trachomatis and N. gonorrhoeae by real-time PCR in the 3 types of specimens were compared to those by ProbeTec ET assay (ProbeTec, BD Diagnostic System). The overall positive concordance between real-time PCR and ProbTec were 97.1% (66/68) for C. trachomatis and 100% (33/33) for N. gonorrhoeae, C. trachomatis detection yielded 3 discordant results in endocervical specimens and 1 discordant result in male FVU by real-time PCR and ProbTec. Three of 4 reexamined using Aptime Combo 2 Assay (Fuji Rebio Inc.) were positive for C. trachomatis. Endocervical swab and FVU specimen results for C. trachomatis were discordant in 3 cases in real-time PCR and 4 in ProbeTec. Subjects with 2 or more positive endocervical awab results in female or male FVU specimens were assumed to be "true positive" for C. trachomatis. The sensitivities of real-time PCR for detecting C. trachomatis was 94.4% in endocervical swabs, 77.8% in female FVU and 97.4% in the male FVU. The sensitivities for real-time PCR for detecting N. gonorrhoeae was 100% in all 3 specimentypes. Abbott RealTime CT/NG assay was useful for detecting C. trachomatis using endocervical swabs or male FVU specimens and for detecting N. gonorrhoeae using endocervical swabs and all FVU specimens.