深圳地区泌尿生殖道解脲支原体和沙眼衣原体感染对妊娠的影响

采用聚合酶链反应（ＰＣＲ）技术，对不孕妇女７９人进行生殖道解脲支原体（ＵＵ）及沙眼衣原体（ＣＴ）检查，结果ＵＵ－ＤＮＡ阳性５６人占（７０．９％），ＣＴ－ＤＮＡ阳性２６人（占３３％），其中１８人ＵＵ－ＤＮＡ和ＣＴ－ＤＮＡ均为阳性。这些患者在治疗不孕症过程中首先治愈ＵＵ或ＣＴ感染，其治疗效果与５１例同类病例进行比较，治愈率有明显提高，差异有极显著意义（Ｐ＜０．０１），结果提示，泌尿生殖道解脲支原体及沙眼衣原体感染与女性不孕症有密切的关系，应引起临床医师的重视。     

PCR检测支原体和沙眼衣原体在产科中的应用

采用常规聚合酶链反应（ＰＣＲ） 和套式ＰＣＲ（ＮＰＣＲ） 技术检测妊娠２１ ～３１ 周１９０ 份孕妇子宫颈分泌物中人型支原体（ＭＨ） 、解脲支原体（ＵＵ）和沙眼衣原体（ＣＴ）ＤＮＡ， 结果表明各感染原ＤＮＡ总阳性率为３７－９％ （７２／１９０） ； 其中ＭＨ、ＵＵ与ＣＴ ＤＮＡ检出率各为６－３％ （１２／１９０）， ２５－３ ％ （４８／１９０ ）与６－３％ （１２／１９０） ； 在ＤＮＡ阳性组中， 支原体与衣原体阳性率分别为８３－３％ （６０／７２） 与１６－７％ （１２／７２ ）。结果提示， 对上述感染者给予及时足量与足程的抗支原体与抗衣原体治疗， 将有利于胎儿的健康生长与发育。     

拟诊为淋病患者中泌尿生殖道沙眼衣原体基因分型及序列分析

目的 了解深圳市拟诊为淋病患者中泌尿生殖道沙眼衣原体的合并感染情况及其基因型分布和序列变异特点.方法 采集401例拟诊为淋病患者的泌尿生殖道分泌物样本,应用Roche Amplicor全自动核酸检测系统对样本进行淋球菌和沙眼衣原体双检,提取DNA,应用巢式聚合酶链反应(nested-PGR)扩增沙眼衣原体主要外膜蛋白基因(omp1)中的VS1～VS2片段,并对其进行序列测定,所获得的序列利用Mega4.0软件与标准参考株进行比对,分析确定其基因型及序列变异情况.结果 401例拟诊为淋病患者中淋球菌的感染率为82.3%(330/401),沙眼衣原体的感染率为24.2%(97/401),淋球菌和沙眼衣原体的合并感染率为21.7%(87/401).97份沙眼衣原体阳性样本中获得73份沙眼衣原体基因片段序列,共检出8个基因型,分别为E型(27.4%)、G/Ga型(23.3%)、D/Da型(16.4%)、F型(13.7%)、J型(11.0%)、H型(5.5%)、B和K型(各1.4%).序列分析发现3例(4.1%)菌株发生错义突变,分别为D/Da型、E型、G/Ga型;F型、H型、J型和K型序列虽多见碱基突变,但均为同义突变.结论淋病患者合并感染沙眼衣原体的比例较高,且泌尿生殖道沙眼衣原体的基因型以E、G/Ga、D/Da和F型为主.序列分析可以为泌尿生殖道沙眼衣原体的分子流行病学研究提供依据.     

聚合酶链反应检测胎盘组织中解脲支原体与沙眼衣原体感染

本文应用聚合酶链反应技术对３８６例产妇的胎盘进行了解脲支原体，沙眼衣原体检测，检出ＵＵ阳性者４０例，阳性率１０．４％，ＣＴ阳性者３２例。阳性率８．３％，提示胎盘组织中ＵＵ，ＣＴ感染与先天性宫内感染有关，值得重视。结果表明ＰＣＲ法具有高度特异性，敏感性，且快速，简便，可有效地检出胎盘组织中ＵＵ、ＣＴ感染。     

聚合酶链反应检测丝虫病患者泌尿道沙眼衣原体及解脲支原体的研究

本文采用聚合酶链反应方法,检测了５０ 例丝虫病患者尿中沙眼衣原体（ＣＴ） 及解脲支原体（ＵＵ）ＤＮＡ,同时选择３０ 例正常人作为对照。结果显示,丝虫病患者尿中ＣＴＤＮＡ、ＵＵ ＤＮＡ 和两者混合感染检出率分别为３０ ％ 、２５ ％ 和２０ ％ ,高于正常对３６３％ （Ｐ＜０－０１）,ＣＴ及ＵＵ 感染与病人的病情、病程有一定的关联。提示乳糜尿的发生、发展与泌尿道ＣＴ 和ＵＵ感染有关。     

深圳地区泌尿生殖道解脲支原体和沙眼衣原体感染对妊娠的 …

采用聚合酶链反应（ＰＣＲ）技术，对不孕妇女７９人进行生殖道解脲支原体（ＵＵ）及沙眼衣原体（ＣＴ）检查。结果ＵＵ－ＤＮＡ阳性５６人占（７０．９％），ＣＴ－ＤＮＡ２６人（占３３％），其中１８人ＵＵ－ＤＮＡ和ＣＴ－ＤＮＡ均为阳性。     

聚合酶链反应检测丝虫病患者泌尿道沙眼衣原体及解脲法原体的研究

本文采用聚合酶链反应方法,检测了５０例患者尿中沙眼衣原体（ＣＴ）及解脲支原体（ＵＵ）Ｄ届时选择３０例正常作作为对照,结果显示,丝虫病患者尿中ＣＴ ＤＮＡ、ＵＵ ＤＮＡ和两者混合感染检出率分别为３０％、２５％和２０％,高于正常对３６３％（Ｐ〈０．０１）,ＣＴ及ＵＵ感染与病人的病情、病程有一定的关联。提示乳糜尿的发生、发展与泌尿道ＣＴ和ＵＵ感染有关。     

孕妇解脲支原体、沙眼衣原体感染的诊断与药物干预

目的探讨妊娠期孕妇解脲支原体（UU）、沙眼衣原体（CT）感染的发病率及其对感染者的治疗效果。方法对108例妊娠妇女采用核酸杂交的方法检测宫颈分泌物,对标本UU、CT-DNA进行DNA扩增、核酸杂交检测。结果 108例妊娠早中期孕妇宫颈分泌物UU、CT的阳性率分别是33.33%,7.41%,口服阿奇霉素治疗后检测UU、CT阳性率分别是17.59%,1.85%。两组间有显著性差异（P=0.001或P=0.002）。结论解脲支原体、沙眼衣原体是妊娠期妇女感染的常见病原体,临床治疗效果显著值得推广应用。     

南充市门诊男性泌尿生殖道标本解脲支原体和沙眼衣原体的感染情况研究

目的分析南充市门诊就诊男性泌尿生殖道标本UU 和CT 感染情况以及年龄分布特点,加强本区域男性CT 和UU 筛查遥方法收集2014 年7 月~2017年7 月共657 例门诊男性尿道分泌物标本,采用荧光PCR 法进行UU尧CT测定,并将所有 男性受检者分为5 个年龄组,对所得检测结果进行统计学分析遥结果657 例受检者共有阳性267 例,总感染率40.64%,单一感 染率36.07%,单一CT 感染14.00%,UU感染22.07%,CT合并UU感染率4.57%遥不同年龄段中21~50 岁三个年龄组感染率最 高（分别44.28%尧41.18%尧43.90%）,21~30尧31~40尧41~50 岁三个年龄组分别与逸51 岁比较,差异有统计学意义（约0.05）遥此外, CT合并UU感染也主要分布在21~50 岁,所有年龄组UU感染阳性率高于CT 感染遥结论南充地区男性CT 和UU的感染 率均较高,以单一感染为主,尤其好发于21~50 岁遥加强CT 和UU 筛查,对性传播疾病防治尧泌尿生殖系感染的诊疗和优生优 育有重要意义遥     

泌尿生殖道支原体、衣原体感染的研究

支原体、衣原体虽可存在于正常人群中 ,但在泌尿生殖道及附件的致病作用愈来愈受到人们的重视。自 195 4年shepard首先从非淋病性尿道炎分泌物中分离解脲脲原体以来 ,相继得到许多学者的证实。衣原体引起泌尿生殖系统感染的发病率已超出淋病感染 ,成为性传播性疾病中最多见的一种病原体我们收集泌尿生殖道炎症、不孕症等方面标本 10 2 0份 ,进行支原体、衣原体分离 ,现就此类微生物的检查情况以及在各种疾病之间的分布报道如下 :1　材料与方法1.1　检测对象　 10 2 0例门诊患者 ,其中非淋病性尿道炎2 12例 ,不孕症 2 2 2例 ,前列腺炎 112例 …     

PCR引物浓度对寡核苷酸液相杂交的影响

目的考察PCR引物浓度配比对液相微珠杂交效率的影响,寻求具有较强杂交信号和较好稳定性的PCR引物浓度配比。方法建立HLA-DRB1等位基因的相关数据库,选择在HLA-DRB1位点的第二外显子上设计探针,并且选择其保守序列作为阳性对照探针(DPC2),DPC2探针中间位点T突变成A作为阴性对照探针(DNC)。分别针对标本C2-008、C2-024、C2-025的等位基因序列设计出6条约21bp的寡核苷酸探针,各探针5’端用氨基(NH2)修饰。通过引物浓度梯度配比(1:100、1:50、1:20、1:8、1:4、1:2、1:1),对型别已知的细胞株DNA进行PCR扩增并得到目的片段(1:100配比除外),在相同条件下将PCR产物与寡核苷酸探针进行液相杂交检测。结果浓度配比为1:1的对称式扩增产物杂交结果不理想,而浓度配比分别为1:20、1:8、1:4、1:2的不对称扩增均得到了待检单链、双链DNA混合物,其中1:4浓度配比具有最好的扩增效率和稳定性。根据阳性信号与阳性标本是否相符表明:引物浓度配比为1:100的不对称PCR和1:1的对称PCR检测效果差,易出现假阴性;1:2、1:4、1:8、1:20配比检测效果较好,比较稳定。结论PCR引物浓度配比影响液相微珠杂交效率,不对称PCR产物有利于提高杂交效率,为快速成功配制PCR试剂奠定了基础,有利于寡核苷酸液相芯片的应用。     

荧光定量PCR检测沙眼衣原体及临床应用

目的：用荧光定量聚合酶链反应（FQ－PCR）检测沙眼衣原体的含量,探讨FQ－PCR在衣原体性尿道炎诊断中的价值。方法：应用荧光探针标记引物的荧光定量聚合酶链反应对172例疑为沙眼衣原体感染患者标本进行检测,并与常规PCR（电泳－EB染色）进行比较。结果：FQ－PCR阳性率为19．8％,常规PCR法为20．9％,两法符合率为63．9％。女性宫颈分泌物标本FQ－PCR阳性为29．2％,常规PCR法为16．7％;男性分泌物FQ－PCR阳性率为16．9％,常规PCR法为13．8％。FQ－PCR特异性较常规PCR法高。结论;FQ－PCR在扩增中实时在线检测,并选择理想的标准曲线做出较精确的临值分析,具有较高的特异性和敏感性,对病原体的诊断有一定的临床意义。     

Multiplex PCR for detection of Chlamydia trachomatis and Neisseria gonorrhoeae in Genitourinary specimens.

We developed a multiplex PCR (M-PCR) assay for the simultaneous detection of Chlamydia trachomatis and Neisseria gonorrhoeae. M-PCR employed C. trachomatis-specific primers KL1-KL2 and N. gonorrhoeae-specific primers HO1-HO3 and produced products of 241 and 390 bp, respectively. PCR products were easily detected by agarose gel electrophoresis and confirmed by Southern hybridization using labelled oligonucleotide probes. M-PCR had a sensitivity of 10 fg of C. trachomatis and N. gonorrhoeae DNA (equivalent to 1 to 2 genome copies). M-PCR detected the presence of C. trachomatis and N. gonorrhoeae DNA in 15 male urethral and 12 female endocervical specimens, 3 of which were positive for C. trachomatis, 18 of which were positive for N. gonorrhoeae and 6 of which were positive for both organisms. M-PCR was evaluated further by testing 200 male first void urine (FVU) specimens, of which 18 were positive by C. trachomatis PCR and Chlamydiazyme and 4 were positive by C. trachomatis PCR but negative by Chlamydiazyme. All 22 FVU specimens were positive by a confirmatory PCR using a second plasmid target and were positive by M-PCR. Ten of 11 men with cultures that were positive for N. gonorrhoeae had FVU specimens that were positive by both N. gonorrhoeae PCR and M-PCR. Two other men with negative N. gonorrhoeae urethral cultures had FVU specimens that were positive by N. gonorrhoeae PCR, by two confirmatory N. gonorrhoeae PCR assays using 165 rRNA and cytosine methyltransferase primers, and by M-PCR. The sensitivity of M-PCR for detecting C. trachomatis was 100% (22 of 22 specimens), compared with 81.8% (18 of 22 specimens) for enzyme immunoassay. Sensitivity of M-PCR for N. gonorrhoeae was 92.3% (12 of 13 specimens) compared with 84.6% (11 of 13 specimens) for urethral culture. The specificity of M-PCR was 100% for both C. trachomatis (178 of 13 specimens) and N. gonorrhoeae (187 of 187 specimens). M-PCR testing of FVU specimens provided a sensitive and noninvasive method for detecting C. trachomatis and N. gonorrhoeae infection in men.     

PCR结合寡核苷酸探针杂交检测临床常见真菌的实验研究

目的 建立PCR结合生物标记的寡核甘酸探针斑点杂交技术,鉴定临床常见的真菌。方法 首先用真菌通用引物扩增白念球菌、热带念球菌、假热带念球菌、近平滑念球菌、光滑念球菌、解脂念球菌、克鲁斯念球菌、季也蒙念球菌、黄曲 霉、烟曲霉的核糖体大亚单位基因的保守区序列,然后用生物素标记的种特异性寡核苷酸探针与扩增产物杂交,并将此方法用于临床标本和临床分离菌株的检测。结果 通用引物可以扩增上述11种临床常见真菌的DNA,扩增片段长度在260bp左右。9种特异性探针分别与11种真菌标准菌株的PCR扩增产物杂交,结果表明每种探针都具有高度特异性。斑点杂交法和Southerm杂交法检测敏感性相同,为100fg;琼脂糖凝胶电泳法检测敏感性为1pg。通过69例临床标本和31例临床分析菌株的检测,PCR－杂交法的结果和真菌培养法的结果基本一致。结论 PCR结合生物素标记的寡核苷酸探针杂交技术可将9种临床常见真菌鉴定到种,方法快速、敏感、特异。     

Duplex PCR system for simultaneous detection of Neisseria gonorrhoeae and Chlamydia trachomatis in clinical specimens.

AIMS--To evaluate the use of a duplex polymerase chain reaction (PCR) assay for the simultaneous detection of Neisseria gonorrhoeae and Chlamydia trachomatis in clinical samples. METHODS--Genital swab specimens were obtained from both China (203 swabs) and Hong Kong (202 swabs). N gonorrhoeae and C trachomatis were detected in each specimen with a number of tests including enzyme immunoassays (IDEIA) and PCR assays using both single and double primer pairs. The primer pair for N gonorrhoeae was derived from the cppB gene on its cryptic plasmid and the PCR product was 390 base pairs long. For C trachomatis, the PCR product was 473 base pairs long, resulting from amplification of a sequence in the common 7.4 kilobase plasmid present in all serovars. For N gonorrhoeae, PCR results were also compared with those obtained by culture and Gram's smear of the discharges. RESULTS--For the 203 specimens collected in China, similar numbers of positive results (177) were obtained by both Gonozyme and duplex PCR for the detection of N gonorrhoeae. No discrepant results were found among the cultured specimens when Gonozyme and duplex PCR were compared. C trachomatis was detected in 47 specimens by duplex PCR, but was detected in only 28 by IDEIA. Of the 202 Hong Kong specimens, 46 were positive for N gonorrhoeae, detected by both Gonozyme and duplex PCR; 34 were positive for C trachomatis, 25 of which were detected by IDEIA and the remainder by duplex PCR. CONCLUSIONS--The duplex PCR assay is a satisfactory diagnostic tool for the simultaneous detection of N gonorrhoeae and C trachomatis in clinical swab samples. Further evaluation is suggested.     

Use of PCR and reverse line blot hybridization assay for rapid simultaneous detection and serovar identification of Chlamydia trachomatis

The aim of this study was to develop and evaluate multiplex and nested PCR-reverse line blot (RLB) hybridization assays for detection and serovar identification of Chlamydia trachomatis. Two sets of primers targeting the VD2 region of the omp1 gene and one set targeting the cryptic plasmid were designed for use in multiplex (both targets) and nested PCR (omp1 only). For the RLB assay, labeled omp1 amplicons were hybridized to a membrane containing probes specific for 15 C. trachomatis serovars. The assays were used to test 429 clinical specimens, which had been previously tested for C. trachomatis using the COBAS AMPLICOR system. Specimens were tested without knowledge of the COBAS AMPLICOR result. Of 205 specimens that were positive by COBAS AMPLICOR, 201 (98%) were positive by multiplex PCR-RLB and 188 (92%) were also positive by omp1 nested PCR-RLB. In addition, three of 224 COBAS AMPLICOR-negative specimens were positive by omp1 nested PCR-RLB. One hundred sixty-six of 191 (87%) specimens in which C. trachomatis serovars were identified contained only one serovar and 25 (13%) contained two or three serovars. Serovars D, E, and F were found in 31 (16%), 83 (43%), and 51 (27%) specimens, respectively. Serovar E (41%) was the most commonly identified single serovar. Serovars J and K were found alone uncommonly (<2% each), but 18 of 25 (72%) specimens with multiple C. trachomatis serovars contained one or both (10 specimens) of these serovars. The nested (ompI) PCR-RLB is a specific and sensitive method for simultaneous detection and serovar identification of C. trachomatis, which can reliably identify mixed C. trachomatis serovars. It is suitable for use in epidemiological studies.     

不孕不育者生殖道淋球菌、沙眼衣原体和解脲支原体的检测结果分析

目的了解在不孕不育患者中生殖道淋球菌（NC）、沙眼衣原体（CT）、解脲支原体（UU）感染的情况。方法应用实时荧光定量PCR方法对465例不孕不育患者生殖道3种病原体基因进行定量测定。结果阳性检出268例，总阳性率为57．64％，其中UU的阳性检出率最高，占43．87％；重叠感染中以UU＋CT的阳性检出率最高，占3．44％。结论不孕不育患者生殖道中3种病原体感染率不尽相同，尤以UU感染率最高，是造成不孕不育的重要原因。     

Direct fluorescent antibody assay and polymerase chain reaction for the detection of Chlamydia trachomatis in patients with vernal keratoconjunctivitis

OBJECTIVES:

To identify Chlamydia trachomatis via polymerase chain reaction and a direct fluorescent antibody assay in patients with vernal keratoconjunctivitis while comparing the efficacies of both tests for detecting Chlamydia trachomatis in these conditions.

METHODS:

Conjunctival scraping samples were obtained from 177 patients who were divided into two groups: a vernal keratoconjunctivitis group (group A) and a control group (group B). The polymerase chain reaction and a direct fluorescent antibody assay were performed. Sensitivity, specificity, receiver operating characteristic curves, and areas under the curve were calculated for both tests in groups A and B. Receiver operating characteristic curves were plotted using a categorical variable with only two possible outcomes (positive and negative).

RESULTS:

Statistical analysis revealed a significant association between vernal keratoconjunctivitis and Chlamydia trachomatis infection detected by a direct fluorescent antibody assay with high sensitivity and specificity. All patients in group A with positive polymerase chain reactions also presented with positive direct fluorescent antibody assays.

CONCLUSION:

The association between vernal keratoconjunctivitis and Chlamydia trachomatis infection was confirmed by positive direct fluorescent antibody assays in 49.4% of vernal keratoconjunctivitis patients and by positive polymerase chain reactions in 20% of these patients. The direct fluorescent antibody assay detected Chlamydia trachomatis in a higher number of patients than did the polymerase chain reaction. Although the diagnosis of trachoma is essentially clinical, the disease may not be detected in vernal keratoconjunctivitis patients. Due to the high frequency of chlamydial infection detected in patients with vernal keratoconjunctivitis, we suggest considering routine laboratory tests to detect Chlamydia trachomatis in patients with severe and refractory allergic disease.     

聚合酶链反应-反向杂交快速检测巨细胞病毒糖蛋白gB、gH基因型变异

目的 建立一种简便、快速、敏感和特异的检测巨细胞病毒（HCMV）糖蛋白gB、gH基因型变异的方法。方法 以HCMV糖蛋白基因gH、gBn、gBclv为靶序列，设计17条具有型特异性的寡核苷酸探针，聚合酶链反应(PCR)扩增目的片段，反向杂交检测23例中国和6例德国移植患者HCMVgH、gBn、gBclv基因型变异，其结果与直接测序进行比较。结果 23例中国患者HCMV糖蛋白为gH1和2型，gB1、2和3型，未见gB4型；而6例德国患者的HCMV糖蛋白涵盖gH1和2型，gB1、2、3及4型。中、德国两患者均可同时感染2种不同基因型的病毒株。反向杂交技术对检测患者同时感染不同基因型病毒株优于直接测序。结论 PCR－反向杂交技术具有简便、快速、敏感和特异的特点，适用于临床实验室检测HCMV糖蛋白基因型变异。