47例慢性非细菌性前列腺炎患者前列腺液细菌学检测结果分析

慢性前列腺炎（CP）易反复发作,病原体检测困难。以往临床常采用前列腺液细菌培养检测病原菌,但受条件限制,其检出率不高。2004年～2006年5月。我们采用分子生物学技术对51例慢性非细菌性CP患者行前列腺液标本细菌学检测,现分析结果。     

性传播疾病后慢性前列腺炎68例报告

近年随着性传播疾病（STD）的发病率升高,性病后前列腺炎的发病率亦不断增加。特对2002年9月～2004年11月诊治68例STD后前列腺炎患者的前列腺液病原学检测结果进行分析,并讨论发病因素与治疗,报告如下。     

聚合酶链反应检测幽门螺杆菌70例报告

聚合酶链反应检测幽门螺杆菌７０例报告李延年，刘先贤（武汉市东西湖区人民医院消化内科武汉４３００７１）１９９０年Ｈｏｓｈｉｎａ等首先应用聚合酶链反应（ＰＣＲ）成功检出胃粘膜活检标本内的ＨＰ。我院从今年１９９年１月份开始，在短短三个月中对７０例胃镜受检者...     

直肠粘膜下注入法治疗慢性前列腺炎（附300例报告）

1988-03／2002-10我们采用直肠粘膜下穿刺注射法，将药物直接注入前列腺及其周围组织，治疗300例慢性前列腺炎病人，效果满意，报告如下。     

老年前列腺炎的特点和处理(附35例报告)

老年前列腺炎,临床并不少见,于2001年2月至2002年12月,诊治35例,报告如下.     

多虑平治疗ⅢB型难治性前列腺炎疗效观察（附35例报告）

目的观察多虑平治疗ⅢB型慢性难治性前列腺炎的疗效。方法将65例ⅢB型慢性难治性前列腺炎患者随机分为观察组（35例）和对照组（30例）,分别采用多虑平和传统方法治疗。观察两组疗效和慢性前列腺炎症状积分指数（NIH—CPSI）评分。结果治疗后两组NIH—CPSI评分均低于治疗前,观察组低于对照组（P均〈0．01）。观察组和对照组治疗有效率分别为82．86％和36．67％（P〈0．01）。两组均无严重不良反应发生。结论采用多虑平治疗ⅢB型慢性难治性前列腺炎疗效好、安全。     

前列腺被膜十字切开术治疗慢性前列腺炎：附35例报告

对３５例慢性前列腺炎行前列腺被膜十字切开术２为慢性前列腺炎的治疗提供了一个新手术方法。     

应用双重式聚合酶链反应技术检测鼠疫菌的探讨

聚合酶链反应(PCR)是20世纪80年代发展起来的分子生物学技术。随着Taq酶的应用和自动化扩增仪的发展，PCR技术迅速渗透到生命科学的各个领域。张景林报告Hinnebush等(1993)用Pla基因序列，以特异性片段为引物，建立了检测印鼠客蚤体内鼠疫菌的PCR法，Hiroko氏等(1996)用多引物PCR对鼠疫菌检测进行研究；吴明寿等     

实时定量多聚酶链反应技术在诊断曲霉菌感染中的应用

近年曲霉菌感染的发病率急剧上升,曲霉病的病死率亦高居不下。早期诊断曲霉菌感染是改善预后的关键。曲霉病临床表现无特异性,早期诊断困难。最新的实时定量聚合酶链反应技术集快速、灵敏、特异、定量等优点于一体,在曲霉菌感染的诊断中有着广阔的应用前景。     

聚合酶链反应诊断军团菌感染的实验研究

目的早期诊断军团菌感染。方法采用聚合酶链反应（ＰＣＲ）检测嗜肺军团菌（Ｌｐ）ｍｉｐ基因的特异性片段，并应用于豚鼠米克戴德军团菌（Ｌｍ）感染后的组织标本检测。结果可以扩增出ＬｐＩ型和Ｌｍ６３０ｂｐ的特异性片段，其他临床分离株不被测得，检测灵敏度为１５０ｆｇ军团菌基因组ＤＮＡ（相当于１５～３０个军团菌）。对２２份Ｌｍ感染后不同时间获取的豚鼠组织标本，应用该法与培养法检测结果比较显示，在感染后３．５天培养法阳性率为８８．９％，ＰＣＲ法１００％，在感染后７．５天培养法阳性率为０，而ＰＣＲ法仍有２２．２％的阳性率。结论本方法敏感、特异，既能检出嗜肺军团菌，又能测得Ｌｍ，大大提高了军团病的诊断率     

应用多重聚合酶链式反应鉴别布鲁杆菌

目的 探讨应用多重聚合酶链式反应(Muhiple-PCR)进行布鲁杆菌株种型的鉴别.方法 以6株布鲁杆菌标准菌株(牛种、羊种、猪种、犬种、绵羊附睾种、沙林鼠种布鲁杆菌标准菌株)作为阳性对照,以大肠埃希菌O∶157和小肠结肠炎耶尔森菌O∶9作为阴性对照,待测布鲁杆菌株29株.先用布鲁杆菌属特异性聚合酶链式反应(BCSP31-PCR)扩增上述待测布鲁杆菌株,扩增结果为阳性的菌株再用Muhiple-PCR方法进行布鲁杆菌种型鉴别.结果 29株待测布鲁杆菌株BCSP31-PCR方法扩增均为阳性,进一步Multiple-PCR方法扩增均为阳性,其中有20株鉴定为羊种菌,5株鉴定为猪种菌、3株鉴定为牛种菌、1株鉴定为犬种菌.结论 Multiple-PCR方法是一种快速、特异、简便、低风险的布鲁杆菌种型鉴定方法.     

巢式聚合酶链反应检测慢性粒细胞白血病bcr／abl融合基因

目的 探讨Ph染色体和bcr/abl融合基因在慢性粒细胞白血病（CGL）的发病机制、诊断、治疗、预后判断的价值。方法 对46例CGL患者作巢式逆转录聚合酶链反应（RT-PCR）检测bcr/abl融合基因,同时对其中28例作细胞遗传学检查。结果 46例CGL患者中,44例bcr/abl融合基因阳性,阳性率为95.7%;28例CGL患者作细胞遗传学检查。26例Ph染色体阳性,阳性率为92.9%;2例Ph染色体阴性的CGL患者,用RT-PCR检测出ber/abl融合基因。结论 巢式RT-PCR是一种快速、敏感而准确的检测方法,可以为部分Ph染色体阴性的CGL患者提供分子生物学的诊断依据。     

Real-time polymerase chain reaction for microbiological diagnosis of parapneumonic effusions in Canadian children

BACKGROUND:

Community-acquired pneumonia (CAP) complicated by parapneumonic effusion/empyema is an infectious syndrome commonly encountered by physicians caring for children in Canada.

OBJECTIVE:

To investigate the incremental benefit of novel molecular testing for the microbiological diagnosis of pediatric CAP complicated by parapneumonic effusion/empyema in Canada.

METHODS:

A convenience sample of pleural fluid from 56 children who had been admitted to hospital in Ontario with CAP complicated by parapneumonic effusion between 2009 and 2011 was examined. Multiple uniplex real-time polymerase chain reaction (PCR) testing was performed on these pleural fluids and compared with traditional culture-based testing of blood and pleural fluid samples.

RESULTS:

Molecular methods detected a pathogen in 82% of cases, whereas traditional cultures of blood and pleural fluids detected a pathogen in only 25%. The majority of parapneumonic effusions were associated with pneumococcal infection; Streptococcus pneumoniae was detected in 62% of the samples using molecular methods but in only 14% of samples using culture-based methods. Streptococcus pyogenes, detected in 16% of samples using PCR, was the second most common pathogen found. No patients were found to have empyema caused by Staphylococcus aureus.

DISCUSSION:

The results showed that multiple uniplex real-time PCR performed substantially better than traditional culture methods for microbiological diagnosis of CAP complicated by effusion/ empyema. S pneumoniae and S pyogenes were found to be responsible for the majority of infections. The approach detected pathogens in a similar proportion of pleural fluid samples as previously reported nested PCR assays; furthermore, the real-time closed-well approach also minimized the risk of nonspecificity due to cross-contamination relative to nested PCR.

CONCLUSIONS:

Real-time PCR for the detection of bacterial DNA in pleural fluids has the potential to better define the microbiological cause of pediatric CAP. This approach could help clinicians provide targeted antimicrobial therapy.     

多重PCR检测方法在鼠疫监测中的应用

目的验证在鼠疫疫源地调查中应用多重PCR方法快速检测鼠疫菌的实用性。方法在14个省的17个鼠疫监测点采集标本2524份。选用针对鼠疫菌特异序列的引物,并加入内部对照模板,直接从鼠肝、脾等脏器标本中进行鼠疫核酸检测,与细菌分离培养结果相比较并进行统计学分析。结果两种检测方法的符合率为92.67%。PCR方法阳性检出率为10.38%,较细菌培养阳性检出率(4.48%)高(x~2=682.25,P<0.01)。结论多重PCR方法可应用于鼠疫监测中,比传统方法迅速、灵敏,但还有待于优化。     

Validity of the polymerase chain reaction in the diagnosis of clinically suspected cases of American visceral leishmaniasis

To test the validity of the polymerase chain reaction for diagnosing American visceral leishmaniasis, 88 suspected cases were studied. Diagnosis was confirmed in 47 (53.5%) and ruled out in 41 (46.5%) patients. Samples of bone marrow and peripheral blood were processed by polymerase chain reaction to evaluate the sensitivity and specificity of the test and its agreement beyond chance with microscopy examination. The polymerase chain reaction was positive in bone marrow of 100% of the patients with amastigotes seen with microscopy examination, and in 59.5% in those where no parasite were seen. Agreement beyond chance between visualization of the parasite in bone marrow aspirates and polymerase chain reaction was considered weak (Kappa = 0.41). Concordance between polymerase chain reaction of bone marrow aspirates and of peripheral blood was considered excellent (Kappa = 0.88). The test turned out positive in all bone marrow aspirates of those with the disease and whereas the positivity rate was 58.5% among those without the disease, with specificity rate of 41.5%.     

Rapid detection of chimeric bcr/abl mRNAs in acute lymphoblastic and chronic myeloid leukemia by the polymerase chain reaction

Summary We have developed a rapid method for the detection of bcr/abl mRNAs, the products of the BCR/ABL fusion genes. The method is based on the polymerasechain-reaction (PCR). Through the use of additional internal primers it is possible to detect directly a single Ph¹-positive cell among 10⁵ unaffected cells thus omitting time-consuming blotting procedures. The whole analytical procedure starting from RNA isolation to agarose gel electrophoresis including two rounds of PCR can be performed in less than six hours.Supported by the Wilhelm Sander-Stiftung, Neustadt/Donau and the Deutsche Krebsgesellschaft, Landesverband Berlin     

实时定量聚合酶链反应技术在鉴别结节病与增殖性结核病中的应用

目的利用实时定量聚合酶链反应（PCR）方法检测结节病和结核病病理组织中结核分枝杆菌DNA，以探讨结核分枝杆菌在结节病发病中的作用及本方法在结节病与增殖性结核病鉴别中的应用价值。方法以上海市肺科医院1998年1月至2003年12月住院患者76例为研究对象，其中结核病组30例、结节病组31例和正常对照组（其他疾病）15例，利用定量PCR检测用石蜡包埋的淋巴结或肺组织中结核分枝杆菌DNA，并以胎鼠肺组织作为阴性对照。结果结核病组结核分枝杆菌DNA检出的阳性率（30／30）明显高于结节病组（6／31）和正常对照组（2／15），结节病组与正常对照组结核分枝杆菌DNA检出的阳性率无明显差别。结核病组结核分枝杆菌DNA检出的绝对和相对拷贝数明显高于结节病组和正常对照组，结节病组与正常对照组无明显差别，而胎鼠肺组织中未检出结核分枝杆菌DNA。结论本研究结果不支持结核分枝杆菌感染与结节病的相关性。实时定量PCR法检测石蜡包埋病理组织中结核分枝杆菌DNA可作为鉴别增殖性结核和结节病的方法之一。     

应用多重聚合酶链反应技术快速鉴定牛分支杆菌卡介苗株

目的 建立快速鉴定牛分支杆菌卡介苗(BCG)菌株的方法。方法 依据最近研究发现的牛分支杆菌BCG菌株不存在命名为缺失区1(deleted region 1，RD1)的基因区，而其他牛分支杆菌菌株和其他结核分支杆菌复合群(MTC)菌种(结核分支杆菌、非洲分支杆菌和田鼠分支杆菌)存在RD1基因区的遗传学信息，应用多重聚合酶链反应(PCR)检测RD1基因区存在与否，以鉴别BCG菌株与其他牛分支杆菌菌株及其他MTC菌种。结果 所试5株BCG疫苗生产用标准菌株和近期国内发生的1例小儿BCG接种后全身播散性感染致死病例的2株BCG分离菌株，均缺失RD1基因区；其他牛分支杆菌菌株，包括3株牛分支杆菌标准菌株、5株分别从结核病牛或鹿分离的牛分支杆菌菌株，1株从结核病患痰标本分离的牛分支杆菌菌株，以及其他MTC菌种，包括结核分支杆菌H37Rv和H37Ra标准菌株、48株从结核病患痰标本分离的结核分支杆菌菌株、3株非洲分支杆菌标准菌株，均存在RD1区。结论子多重PCR技术用于牛分支杆菌BCG菌株的鉴定简便、快速、特异，适于在临床实验室应用。