套式PCR鉴定精液解脲脲原体培养结果报告

应用套式（Ｎｅｓｔｅｄ）ＰＣＲ鉴定７０例精液解脲脲原体培养结果。显示两种方法的阳性符合率为７８．０４％，其中包括：１．培养结果２４－４８小时阳性，而后转阴性１３例，经套式ＰＣＲ检测６例阳性，阳性率为４６．１％，占阳性经率的１６．２％（６／３７）。２．培养７２小时后才显示阳性结果６例，经套式ＰＣＲ检测均为阳性，占阳性比率的１６．２％（６／３７），说明培养法只能用于初筛。     

液体培养法与套式PCR法检测解脲脲原体比较研究

目的：评价解脲脲原体（Ｕｒｅａｐｌａｓｍａ ｕｒｅａｌｙｔｉｃｕｍ,Ｕｕ）液体培养技术的准确性。方法：应用被认为是目前检测临床标本中支原体最为灵敏、特异的套式ＰＣＲ技术,并辅以ＤＮＡ全自动测序技术,对１５４例男性ＳＴＤ者的尿道拭子和８３例女性ＳＴＤ者的阴道拭子配对进行Ｕｕ液体培养法和套式ＰＣＲ法检测。并以套式ＰＣＲ检测为参照系进行统计分析。结果：一、男性ＳＴＤ者Ｕｕ液体培养准确率为６３．０％,假阴性和假阳性率分别为２８．５７％和８．４４％;二、女性ＳＴＤ者Ｕｕ液体培养准确率仅为３８．５５％,假阴性和假阳性率分别４９．４０％和１２．０５％。三、男、女性ＳＴＤ者合并观察Ｕｕ液体培养准确率为５４．４３％,假阴性和假阳性率分别为３５．８６％和９．７％。四、Ｕｕ套式ＰＣＲ产物经ＤＮＡ全自动测序与Ｕｕ标准株（９６０＾Ｔ）完     

新生儿疟眼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）和解脲解原体（ＵＵ）与新生儿疾病的关系，我们采用套式聚合酶链反应（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例，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，     

解脲脲原体感染与早产儿关系研究

本文随机对我院新生儿病房１９９７年６月１９９７年１２月收住的１２３例新生儿患者用聚合酶链反应技术（ＰＣＲ）在入院３天内对其咽拭子标本进行解脲脲原体（ＵＵ）之ＤＮＡ检测。结果：（１）早产儿组ＵＵ阳性率５６５２％显著高于足月儿组ＵＵ阳性率３４００％（ｘ２＝４．０１,Ｐ＜０．０５）。（２）ＵＵ阴性组平均体重３０８２２±３３５１ｇ显著高于ＵＵ阳性组平均体重２７５４３±３１６２ｇ（ｔ＝２．０５１,Ｐ＜０．０５）。结论：宫内感染ＵＵ是引起早产儿、低出生体重儿的重要因素之一。预防关键在对生殖道ＵＵ阳性妇女怀孕前先予治疗,以阻断母婴垂直传播。     

IST支原体培养与PCR解脲支原体检测方法的比较

目的：比较分析ＰＣＲ解脲支原体检测法与ＩＳＴ支原体培养法的结果及两种方法的优缺点。方法：同时采集８８例临床上诊断为非淋菌性尿道炎（ＮＧＵ）、阴道炎患者分泌物标本两份,分别用ＩＳＴ支原体培养法和ＰＣＲ解脲支原体（Ｕｕ）检测法进行对照实验和分析。结果：ＩＳＴ支原体培养法Ｕｕ阳性２５例（２８．９４％）,人型支原体（Ｍｈ）１例（１．１４％）,Ｕｕ＋Ｍｈ５例（５．６８％）。ＰＣＲ检测Ｕｕ阳性３０例（３４．０４％）,其中男性５例ＰＣＲ检测Ｕｕ为阳性,而ＩＳＴ支原体培养法为阴性;女性３例ＰＣＲ检测Ｕｕ阴性,培养法为阳性。两种方法的Ｕｕ阳性率经统计学ｘ２检验（Ｐ＞０．０５）无显著性差别。结论：ＩＳＴ支原体培养法与ＰＣＲ检测Ｕｕ法均能适用于临床Ｕｕ的检测。ＩＳＴ支原体培养法操作简便,不需要特殊仪器;能进行支原体的种类鉴别;有参考病理阈值并可同时做药敏实验,特别适合基层医院的支原体检测工作。     

PCR技术检测非淋菌性尿道炎患者精液中沙眼衣原体及解脲脲原体

沙眼衣原体（ＣＴ）及解脲脲原体（ＵＵ）是非淋菌性尿道炎中的两种主要病原体，其对生殖道的危害及引起的不孕不育已被国内外学者广泛关注。我们对３４例非淋菌性尿道炎患者分别以精液及尿道拭子为标本，以ＰＣＲ技术检测，结果精液标本中ＣＴ及ＵＵ检出的总阳性率为６７．６％（２３／３４），同时检出滴虫５例，霉菌２例。而尿道拭子标本总阳性率为３５．２９％（１２／３４）。未检出滴虫及霉菌，我们认为以精液为标本，取材方便，阳性检出率高。     

男性生殖系感染者精液解脲支原体感染的检测研究

目的：探讨男性生殖系感染者精液解脲支原体（ＩＵＩ）感染的检测研究。方法：取精液标本检测，以源于解脲支原体尿素酶基因序列为引物，应用聚合酶链反应（ＰＣＲ）技术对解脲支原体ＤＮＡ进行扩增。结果：所试几种非解脲支原体培养物均阴性。解脲支原体阳性出现１条４１８ｂｐ的ＤＮＡ扩增带。３８２４例男性生殖道感染者，ＵＵ阳性检出率２９．１％。其中急性淋菌性生死道炎感染率为４８．１％，淋菌后生殖道炎感染率为３０．２％     

输卵管妊娠患者宫颈拭子解脲脲原体套式(Nested)PCR检测

近年来，有关解脲脲原体（ＵＵ）感染与盆腔炎症性疾病（ＰＩＤ）的关系报道较多。但生殖道ＵＵ感染与输卵管妊娠的关系研究报道甚少。为此，我们应用更为灵敏、特异的套式（Ｎｅｓｔｅｄ）ＰＣＲ技术检测了１７例输卵管妊娠患者的宫颈拭子ＵＵ－ＤＮＡ，并以同期５３６例非输卵管妊娠妇科住院病人宫颈拭子为对照组。检测结果为，前者阳性率为７６．５％，而后者仅为２１．４％，有显著性差异（Ｐ＜０．０５）。作者认为ＵＵ感染所致的输卵管急慢性炎症是输卵管妊娠的重要原因。作者建议：婚龄妇女应定期作ＵＵ生殖道感染检查，阳性者及时治疗，以防止ＰＩＤ和输卵管妊娠的发生。     

PCR技术检测非淋菌性尿道炎患者精液中沙眼衣原体及解脲腺 …

沙眼衣原体（ＣＴ）及解脲脲原体（ＵＵ）是非淋菌性尿道炎中的两种主要病原体，其对生殖道的危害及引起的不孕不育已被国内外学者广泛关注。我们对３４例非淋菌性尿道炎患者分别以精液及尿道拭子为标本，以ＰＣＲ技术检测，结果精液标本中ＣＴ及ＵＵ检出的总阳性率为６７．６％（２３／２４），同时检出滴虫５例，霉菌２例。而尿道拭子标本总阳性率为３５．２９％（１２／３４），未检出滴虫及霉菌，我们认为以精液为标本，取材方便     

解脲支原体的PCR快速检测技术在临床上的应用

采用聚合酶链反应（ＰＣＲ）技术扩增解脲支原体４２９ｂｐＤＮＡ片段靶基因，检测２０９６例临床疑为非淋菌感染，阴道炎病人宫颈分泌物，检出阳性７８４例，阳性率为３７．４０％。     

Comparison of in-house and commercial sample preparation and PCR amplification systems for detection of human immunodeficiency virus type 1 DNA in blood samples from Tanzanian adults.

This study compared the performance of several in-house nested PCR systems and the Amplicor human immunodeficiency virus type 1 (HIV-1) PCR kit in the detection of HIV-1 DNA in Tanzanian samples prepared by two different methods. All six of the in-house primer sets evaluated had a higher sensitivity for HIV DNA detection in samples prepared by the Amplicor PCR sample preparation method than in those prepared by the Ficoll-Isopaque (FIP) density gradient centrifugation method. A sensitivity of 100% was achieved by combining two in-house primer sets. The sensitivity of the standard Amplicor HIV-1 PCR kit was only 59%, whereas a modified Amplicor HIV-1 PCR test had a sensitivity of 98%. Our data show that Tanzanian samples prepared by the Amplicor preparation method are more suitable for HIV-1 PCR testing than samples prepared by the FIP method. The modified, but not the standard, Amplicor HIV-1 PCR kit provides an alternative to the nested in-house PCR technique for the diagnosis of HIV infection.     

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）。     

淋病奈瑟菌感染的套式PCR法基因诊断研究

本文根据已发表的淋病奈瑟菌之隐匿性质粒ｃｐｐＢ基因序列，建立了淋病奈瑟菌感染的套式ＰＣＲ基因诊断法。本法灵敏度较普通ＰＣＲ高２－３个数量级，并且特异性更强。灵敏、特异、简便的套式ＰＣＲ法检测，在含菌量较少的男性慢性淋病患者和无症状性的女性淋病患者的诊断中可能有着较高的应用价值。     

A study of tubercular lymphadenitis: a comparison of various laboratory diagnostic modalities with a special reference to tubercular polymerase chain reaction

Objective: The purpose of our study was to compare various laboratory diagnostic methods, namely histopathological examination, Ziehl-Neelsen (ZN) stain, AFB culture by conventional Lowenstein–Jensen (LJ) method and fluorescence-based mycobacterial growth indicator tube (MGIT) technique and polymerase chain reaction (PCR) in clinically suspected cases of tubercular lymphadenitis. Materials and Methods: A total of 65 lymph nodes biopsied from patients clinically suspected of having tubercular lymph nodes were included. Specimens were processed for AFB culture after NaOH-NALC concentration and inoculation on LJ medium and using the MGIT system. PCR was performed on all specimens using a commercial nested PCR kit targeting IS6110 insertion element of Mycobacterium tuberculosis complex. All lymph node specimens were subjected to histopathological examination. Results: Of the 65 lymph nodes, 37 (56.9%) were positive on MGIT culture and 45 (69.2%) were positive by PCR. Histopathology showed maximum sensitivity (96%) but with compromised specificity (78.5%). PCR showed 90.1% sensitivity and 100% specificity. The mean turnaround time for mycobacterial growth in smear negative specimens was 30 days determined by LJ and 20 days by MGIT techniques. Conclusion: PCR is a rapid and useful method for diagnosis of TB lymphadenitis and definitely increases the positive predictive value of a positive histopathology report. MGIT is better than LJ culture as regards time to positivity and higher yield.     

Performance of Bordetella pertussis IS481 real-time PCR in a vaccine trial setting

A real-time PCR method targeting the Bordetella pertussis IS481 gene fragment was evaluated in a vaccine trial setting in which real-time PCR results could be validated against culture and serology results. Two commonly used DNA extraction methods, Amplicor Respiratory Preparation kit and the QIAamp DNA Mini Kit, were compared. An approximately 50-fold higher sensitivity was achieved using the Amplicor kit. 89 of 276 aspirates analysed with the IS481 real-time PCR were positive. Interestingly, six of these were culture negative and came from serology-negative patients. Defining true positive cases either as culture-positive or as PCR-positive cases that had been confirmed with a serology-positive result or verified with a newly constructed recA PCR, the sensitivity and specificity of the IS481 real-time PCR were 89% and 98%, respectively. This study confirms the specificity and high diagnostic sensitivity of IS481-based PCR methods for diagnosis of B. pertussis.     

套式（Nested）PCR检测肺炎支原体与肺炎衣原体研究

本文报告根据肺炎支原体（Mp）和肺炎衣原体（Cpn）的16SrRNA基因序列，建立了套式（Nested）PCR检测方法。灵敏度试验表明，Mp套式PCR和Cpn套式PCR灵敏度分别高出Mp普通PCR和Cpn普通PCR二个数量极。     

Importance of sample preparation for molecular diagnosis of lyme borreliosis from urine

Urine PCR has been used for the diagnosis of Borrelia burgdorferi infection in recent years but has been abandoned because of its low sensitivity and the irreproducibility of the results. Our study aimed to analyze technical details related to sample preparation and detection methods. Crucial for a successful urine PCR were (i) avoidance of the first morning urine sample; (ii) centrifugation at 36,000 x g; and (iii) the extraction method, with only DNAzol of the seven different extraction methods used yielding positive results with patient urine specimens. Furthermore, storage of frozen urine samples at -80 degrees C reduced the sensitivity of a positive urine PCR result obtained with samples from 72 untreated erythema migrans (EM) patients from 85% in the first 3 months to <30% after more than 3 months. Bands were detected at 276 bp on ethidium bromide-stained agarose gels after amplification by a nested PCR. The specificity of bands for 32 of 33 samples was proven by hybridization with a GEN-ETI-K-DEIA kit and for a 10 further positive amplicons by sequencing. By using all of these steps to optimize the urine PCR technique, B. burgdorferi infection could be diagnosed by using urine samples from EM patients with a sensitivity (85%) substantially better than that of serological methods (50%). This improved method could be of future importance as an additional laboratory technique for the diagnosis of unclear, unrecognized borrelia infections and diseases possibly related to Lyme borreliosis.     

Evaluation of nested and real-time PCR assays in the diagnosis of candidaemia

Polymerase chain reaction (PCR) assays have a very low theoretical detection threshold and are therefore advocated for the diagnosis of fungaemia. However, their effectiveness in this respect remains to be assessed. This study compared real-time PCR ( Can -G) and nested PCR assays with blood culture for the diagnosis of Candida spp. bloodstream infections. A total of 200 clinical blood samples obtained from 110 patients at risk for developing a systemic fungal infection, hospitalized in the University Hospital of Sfax (Tunisia), were submitted to testing by culture, nested PCR and real-time PCR. Blood culture was positive in 36 patients. When compared with culture, the Can -G assay (81% sensitivity, 96% specificity) performed better than the nested PCR assay (86% sensitivity, 54% specificity). The real-time PCR assay, which avoids both the contamination hazard with amplicons that may cause false-positive results and the use of time-consuming post-PCR steps, appears more suitable than the nested PCR assay for the laboratory diagnosis of Candida spp. bloodstream infections. In this study, real-time PCR did not enhance the diagnostic sensitivity for Candida spp. bloodstream infections compared with conventional blood culture; however, it may lead to earlier implementation of an adequately targeted antifungal treatment.