实时荧光探针定量PCR在小儿沙眼衣原体肺诊断中的临床应用研究

目的 观察荧光定量PCR在小儿沙眼衣原体肺炎诊断中的应用价值及探讨不同年龄阶段小儿沙眼衣原体(CT)肺炎的临床特点.方法 将我院2004年1～12月份收治60例小儿CT肺炎做研究对象,根据年龄分组,对各组患儿均通过咽拭子或吸痰法获取标本,进行CT-DNA定量分析;回顾性总结分析60例CT肺炎.结果 60例标本中,106 拷贝数5例,105拷贝数6例,104拷贝数10例,103拷贝数39例;小婴儿CT肺炎往往无发热性肺炎,易合并细菌感染;3岁以上儿童则易出现发热、肺部体征大多阴性,而X线多表现为节段性肺炎,其临床经过类似支原体肺炎.结论 1.实时荧光定量PCR检测CT-DNA是诊断小儿沙眼衣原体肺炎最直接、最客观的指标.2.CT肺炎在不同年龄阶段的临床表现存在显著差异.     

实时荧光探针定量PCR在小儿沙眼衣原体肺炎诊断中的临床应用研究

目的观察荧光定量PCR在小儿沙眼衣原体肺炎诊断中的应用价值及探讨不同年龄阶段小儿沙眼衣原体(CT)肺炎的临床特点。方法将我院2004年1~12月份收治60例小儿CT肺炎做研究对象,根据年龄分组,对各组患儿均通过咽拭子或吸痰法获取标本,进行CT—DNA定量分析;回顾性总结分析60例CT肺炎。结果60例标本中,106拷贝数5例,105拷贝数6例,104拷贝数10例,103拷贝数39例;小婴儿CT肺炎往往无发热性肺炎,易合并细菌感染;3岁以上儿童则易出现发热、肺部体征大多阴性,而X线多表现为节段性肺炎,其临床经过类似支原体肺炎。结论1.实时荧光定量PCR检测CT—DNA是诊断小儿沙眼衣原体肺炎最直接、最客观的指标。2.CT肺炎在不同年龄阶段的临床表现存在显著差异。     

荧光定量PCR对不育症患者解脲支原体和沙眼衣原体感染情况的研究

目的用实时荧光定量PCR(FQ-PCR)定量检测不孕不育患者解脲支原体(UU)、沙眼衣原体(CT)基因,观察UU、CT感染情况与不孕不育的关系,探讨实时荧光定量PCR在不孕不育患者诊断和治疗中作用。方法采用FQ-PCR技术检测标本402份。结果 UU阳性率是32.8%,其中男性UU阳性率是21.4%,女性UU阳性率是44.3%。CT阳性率是16.2%,其中男性CT率是11.4%,女性CT率是20.9%。UU感染率在男性、女性患者有极显著性差异(P<0.01),而CT感染率在性别之间有显著性差异(P>0.05)。结论解脲支原体(UU)、沙眼衣原体(CT)是引起男女不育的主要原因之一,实时荧光定量PCR检测UU、CT基因具有操作简单、反应时间短、重复性好、结果客观准确、敏感性和特异性好等优点,其结果对临床诊断、治疗有一定指导意义。     

实时荧光定量PCR检测泌尿生殖道感染患者沙眼衣原体的临床应用

沙眼农原体（chlamydiatrachomatis,CT）是一种独立寄生于细胞内的微生物,含DNA和RNA,革兰染色阴性,只能在细胞内繁殖。CT是近年来最常见的性传播疾病病原体之一,主要引起男女泌尿生殖道感染,园此也越来越受到人们的注意。随着荧光定量PCR技术的发展,该技术已广乏运用于临床。     

荧光定量PCR检测稽留流产患者沙眼衣原体和解脲支原体结果分析

目的探讨沙眼衣原体（CT）和解脲支原体（UU）感染与孕妇稽留流产的相关性,以及荧光定量PCR在稽留流产病因检测中的应用价值。方法采用荧光定量PCR检测稽留流产患者宫颈分泌物标本中的CT-DNA和UU-DNA的含量。结果 138例稽留流产患者中72例感染CT或UU,总检出率53.62%,其中CT阳性率21.74%,UU阳性率42.03%,混合感染10.14%。与正常人群对照差异均有统计学意义（P〈0.05）。结论泌尿生殖系统CT和UU的感染与稽留流产有密切相关性,荧光定量PCR对稽留流产患者CT和UU的检测简单、快速、准确。     

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。     

荧光定量聚合酶链反应检测精液中沙眼衣原体

用荧光定量聚合酶链法对126名男性不育患者和60名正常生育男性的精液进行了沙眼衣原体检测,男性不育组阳性率为19.8%,生育组阳性率为5.0%,两组比较有显著差异。     

新生儿肺炎患儿沙眼衣原体PCR检测研究

应用聚合酶链反应（ＰＣＲ）检测沙眼衣原体（ＣＴ）ＤＮＡ，８４例新生儿肺炎患儿的咽拭子中共检出２１例，阳性率２５．Ｏ％。其中剖宫产儿阳性率１６．６％（２／１２）；经产道产儿阳性率２６．４％（１９／７２）；早产、低体重儿阳性率５０％（５／１０）；而对照组２０例健康新生儿无１例检出。提示新生儿是ＣＴ感染的高危群体；其感染ＣＴ以通过孕妇产道获得为主，但也有宫内感染；早产、低体重儿最易被ＣＴ感染，ＰＣＲ检测ＣＴ感染具有快速、敏感、特异的优点，有广阔的应用前景。     

沙眼衣原体套式（Nested）PCR检测研究

本文报告沙眼衣原体（ＣＴ）的套式（Ｎｅｓｔｅｄ）ＰＣＲ检测方法，本方法ＣＴ隐匿性质粒为靶基因，外套引物采用国外学者所报告灵敏度和特异性较高的序列，内套引物自行设计，经方法学考核表明本法灵敏度和特异性极高。１４６例临检标本套式ＣＴ，ＰＣＲ阳性检出率为３６．３％而市售ＰＣＲ试剂盒（其引物序列与本文外套引物相同）阳性检出率仅为４．１％，前者明显高于后者（Ｐ〈０．０１）。     

PCR检测沙眼衣原体与习惯性流产和女性不孕相关性研究

本文应用聚合酶链反应技术（ＰＣＲ），对１２０例不孕和１００例查体妇女的宫颈内膜标本进行沙眼衣原体检测，以了解女性沙眼衣原体感染与自发流产的关系，并与一般细胞培养法进行了比较。结果：不孕妇女中沙眼衣原体感染率为１８．３％，显著高于对照组（８％Ｐ＜０．０５），并且自发流产３次、４次的妇女沙眼衣原体阳性率明显高于对照组（Ｐ＜０．０１）．敏感性也较一般培养法高（Ｐ＜０．０１）。本研究提示：沙眼衣原体感染与自发性习惯性流产关系十分密切，而且ＰＣＲ法在检测沙眼衣原体感染方面较一般培养法更敏感、快速，是早期诊断生殖道沙眼衣原体感染的一项有价值的方法。     

Quality assessment of conjunctival specimens for detection of Chlamydia trachomatis by PCR in children with active trachoma

One component of control programmes to eliminate trachoma is the treatment of Chlamydia trachomatis infection. A diagnosis of trachoma is based on clinical grounds, but the signs of active trachoma do not always correlate with the presence of C. trachomatis. During a therapeutic trial, the level of C. trachomatis infection in children with active trachoma in Guinea and Pakistan was assessed using a qualitative commercially available PCR that targeted the C. trachomatis plasmid. The influence of the quality of specimens on the efficiency of the PCR was investigated using two quantitative real-time PCRs targeting the specific omp1 gene of C. trachomatis and human chromosomal DNA, respectively. C. trachomatis was detected in c. 23% of children (aged 1-10 years) who presented with clinically active trachoma. Controls showed that PCR-related problems did not influence this detection rate. For 14% of the positive samples, C. trachomatis was detected in only one eye, with a significantly lower mean load of bacteria. These results suggest that epidemiological and therapeutic surveys should be conducted by sampling and testing both eyes. Moreover, the high variability of the cell load observed in the conjunctival swabs suggests that the effectiveness of swabbing may be questionable.     

新生儿疾病沙眼衣原体和解脲脲原体感染的PCR检测研究

为探讨沙眼衣原体（CT）和解脲脲原体（UU）与新生儿疾病的关系，我们采用套式聚合酶链式反应（PCR）技术，从100例住院新生儿的结膜拭子中共检出阳性者58例，阳性率58％（其中CT36倒、UU22例）：剖宫产7例（CT4例，UU3例）；经产道分娩51例（CT32例、UU19例）。结果表明：新生儿是CT、UU易感者．可引起新生儿肺炎、宫内感染、早产、低体重、结膜炎、肠炎。采用先进的套式PCR技术检测具有灵敏、特异、简便的优点。     

Unclassified serovars of Chlamydia trachomatis isolated from Japanese infants

Objective: To analyze antigenic and genetic variations of Chlamydia trachomatis among the serovars obtained from Japanese infants.
Methods: The polymerase chain reaction (PCR) was used to amplify a large part of the major outer-membrane protein gene, and restriction fragment length polymorphism (RFLP) was used to identify the serovars of C. trachomatis from nasopharyngeal and conjunctival swabs from Japanese infants and neonates.
Results: The typing of 10 nasopharyngeal isolates gave the following results: seven E, one H, and two unclassified serovars. The typing of seven conjunctival isolates gave the following results: five D, one F, and one unclassified serovar. Reactive patterns of these unclassified strains, determined by PCR-RFLP, to monoclonal antibodies were different from those of 15 reference serovars.
Conclusions: Characterization of unclassified variants will allow more detailed epidemiologic studies of perinatal C. trachomatis infections in Japan.     

Value of papanicolaou smear in detection of Chlamydia trachomatis infection

This study was undertaken to assess the value of Papanicolaou smear for the diagnosis of Chlamydia trachomatis infection. The study was both retrospective (groups I and II) and prospective (group III). Group I consisted of 41 smears with cytomorphological changes proposed by Gupta, Kiviat, or Shiina. Group II was a control group, consisting of 30 cytologically normal smears. All these smears were subjected to specific immunofluorescent (IF) staining under identical conditions to confirm the diagnosis. In group III, 40 consecutive duplicate cervical smears were collected from patients attending the Sexually Transmitted Disease Clinic. One smear was routinely examined, and the specific IF staining was done on the other smear. The results in all the three groups were analysed. It was concluded that Papanicolaou smear is not useful in the detection of Chlamydia trachomatis infection.     

沙眼衣原体诊断技术研究进展

沙眼衣原体（Ct）是一种特殊的病原体,具有与革兰氏阴性细菌相似的细胞壁,含有DNA和RNA两种类型的核酸,严格寄生于宿主细胞内,沙眼衣原体是一种能够通过滤器,以二分裂方式繁殖的原核细胞型微生物.它具有两种形态：在细胞外具有高度传染性的为原体 在细胞内进行复制、无传染性的为始体.它可以引起非淋球菌尿道炎等许多泌尿生殖道相关疾病,近年来其感染率和危害性已超过淋病奈瑟菌而居性传播疾病之首,眼部衣原体侵入人体眼结膜和角膜引起沙眼和包涵体结膜炎,是世界范围致盲的首要病因.约80%的被感染女性无临床症状,感染反复迁移,造成病理改变,可导致复杂的并发症.因此,早期、简便、快速、特异地发现Ct,对临床的诊断,疾病的早期治疗和预防其流行等具有重要的意义.目前,对沙眼衣原体的诊断方法主要有培养法,免疫学法和分子生物学法.     

Association between Chlamydia trachomatis and abnormal uterine bleeding

PROBLEM: The purpose was to identify distinct inflammatory markers in endometrial tissues of women with abnormal uterine bleeding (AUB) and Chlamydia trachomatis infection. METHOD OF STUDY: Archived endometrial specimens from 92 randomly selected premenopausal women with AUB were examined for C. trachomatis using the species-specific monoclonal antibody against major outer membrane protein (MOMP) and for histopathology associated with inflammation. Statistical analyses included single and multiple logistic regression. Diagnostic accuracy was summarized using receiver operating characteristic (ROC) curves. RESULTS: Chlamydia trachomatis was detected in 44 (48%) of 92 AUB specimens. There were statistically significant correlations of positive MOMP with higher counts of plasma cells (P < 0.01), macrophages (P < 0.0001), and lymphocytic foci (P = 0.01). The ROC curve for macrophages was the strongest predictor (area under the curve = 0.82) for C. trachomatis. CONCLUSION: The prevalence of C. trachomatis in women with AUB is under-estimated. Macrophages appear to be a strong marker for the presence of C. trachomatis in the endometrium.