酶联免疫吸附试验两步法在HIV抗体检测中的应用

目的探讨酶联免疫吸咐试验（ELISA）双抗原夹心一步法和二步法检测人类免疫缺陷病毒（HIV）抗体的不同效果。方法采集2005年10月-2008年10月本市中心血站无偿献血者血液标本48463例,用双抗原夹心ELISA一步法和二步法试剂检测,初筛阳性标本,采用免疫印迹法进行确证。结果检测48463份标本,一步法初筛阳性87例,确证阳性4例,假阳性率0．17％;二步法初筛阳性24例,确讧阳性4侧,假阳性率0．05％,两法的假阳性率比较有显著性差异（P〈0．01）。结论酶联免疫吸咐试验二步法检测人类免疫缺陷病毒抗体的效果优于一步法,更适合于人类免疫缺陷病毒检测的初筛试验。     

孕妇感染性指标检测分析

孕产妇一些传染性疾病不断增加，我们检测了我院近两年来产妇门诊乙型肝炎病毒表面抗原、e抗原、抗丙型肝炎病毒抗体、淋球菌培养、梅毒初筛、梅毒特异性试验、史滋病病毒抗体等六项指标，并进行了回顾性统计分析。产妇2845例，孕周15～26w周。结果2845例产妇中HBsAg阳性241例，阳性率8．47％．HBeAg阳性15例，阳性率0．53％．抗HCV阳性16例，阳性率0．56％，梅毒TRUST初筛试验阳性21例，假阳性5例，阳性率0．56％．梅毒特异1PPA试验阳性24例，阳性率0．84％，淋球菌培养阳性4例，阳性率0．14％，抗HIV均为阴性。     

PCR分析与阴道分泌物直接涂片染色结果比较

用PCR法检测阴道分泌物中病原菌结果与阴道分泌物直接涂片染色结果进行比较，主要表现在了3个方面：1．直接涂片找到革兰氏阴性短小杆菌的标本中，淋球菌的PCR阳性率为48％；2．PCP检测淋球菌的阳性率是40．8％，而直接涂片的阳性率是12．1％，且假阳性占2．6％，P＜0．01；3．对有一定临床症状的患者，阴道清洁度为Ⅳ°时，用PCR法和直接涂片染色法对病原菌的检出率为100％，Ⅲ°时为65．3％，Ⅱ°时为42．3％，结果表明：两方法配合使用更有利于病原学诊断。     

乙型肝炎特殊血清学表现模式的探讨

目的 探讨乙型肝炎病毒（HBV）特殊血清学表现模式。方法 A试剂检测到的乙肝特殊模式标本用B、C两种试剂重检；对HBeAg阳性、HBsAg阴性标本用倍比稀释和二步法重做HBsAg；以聚合酶链反应（PCR）检测乙肝特殊模式的HBV DNA。结果 A试剂检测到的乙肝特殊模式10种，145例；分为“12”同时阳性和“3”阳性、“1”阴性两个模式组；用B、C两种试剂复检的结果同A试剂相比存在较大差异；用倍比稀释和二步法检测“35”模式的HBsAg，阳性率分别提高到75．0％和80．0％，与其HBV DNA的阳性率（74．0％）相一致。结论 不同厂家试剂对乙肝特殊血清学模式的检测结果存在差异。“35”模式标本一步法试剂检测HBsAg漏检率高，用倍比稀释或二步法重检可提高HBsAg的阳性率，应结合HBV DNA的检测结果作出综合判断。     

端粒酶在子宫内膜癌组织及其外周血中的表达

目的：探讨端粒酶（telomerase)在子宫内膜组织、外周血中表达与子宫内膜癌发病的关系及其相关因素。方法：采用聚合酶链反应－银染色法（PCR-A)检测端粒酶在32例子宫内膜癌患者和16例正常对照子宫内膜组织及外周血标本中表达情况。结果：32例子宫内膜癌标本,总阳性率94％。外周血表达阳性率62％。癌组织中端粒酶表达明显高于外周血（P＜0．05）。16例正常子宫内膜组织标本中,表达总阳性率为50％（8／16）。其外周血表达总阳性率为25％（6／16）。正常对照两种标本端粒酶表达未显著水平（P＞0．05）。癌患者两种标本总阳性率均明显高于对照组（P＜0．05）。结论：端粒酶表达与子宫内膜癌的发生有密切的关系,同时也受子宫内膜周期性变化的影响,有可能成为肿瘤的标记物。     

绍兴地区淋球菌PCR检测的临床观察

本文应用聚合酶链反应（ＰＣＲ）技术，对１３００５例临床疑为淋球菌感染患者进行检测，检出阳性６０２例，阳性率为４．６３％。结果表明：ＰＣＲ具有高度的特异性、敏感性，且快速、简便，是临床处理大批标本的好方法，值得临床推广     

育龄妇女生殖道感染病原学研究

目的：了解江苏苏南地区育龄妇女生殖道感染的病原学状况。方法：收集279例育龄妇女生殖道感染者的阴道拭子标本进行支原体（Un,Mh,Mpn,Mg4种）、淋球菌、念珠菌、滴虫和沙眼衣原体检测。结果：患者支原体、淋球菌、念珠菌、滴虫、沙眼衣原体阳性率依闪为75．6％,58．8％、44．1％、32．3％、10．0％,并且合并感染状况严重。结论：支原体、淋球菌、念珠菌、滴虫、沙眼衣原体是江苏苏南地区育龄妇女生殖道感染的常见病原体群,宜采用联合检查,联合治疗的方法加以控制。     

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。     

应用荧光定量PCR法检测病理组织中幽门螺旋杆菌感染

目的探讨荧光定量PCR方法检测病理组织中幽门螺旋杆菌（Hp）的效果,为临床Hp感染诊断提供快速有效的检测方法。方法应用荧光定量PCR法及苯胺蓝染色法分别对106例临床送检胃窦黏膜组织标本进行幽门螺旋杆菌检测,对检测结果进行对比分析。结果在106例胃窦标本中,荧光定量PCR法检测阳性例数68例,阳性率为64．2％,苯胺蓝染色法检测阳性例数46例,阳性率为43．4％。两种方法检测阳性率比较有显著性差异（P〈0．05）。结论荧光定量PCR技术用于检测病理组织中幽门螺旋杆菌,具有灵敏度高、特异性强的特点,对确诊胃镜活检标本中幽门螺旋杆菌感染具有重要应用价值。     

比较两种方法在新生儿苯丙酮尿症筛查中的应用

目的比较串联质谱（tandemmassspectrometry,TMS）技术与茚三酮荧光法在苯丙酮尿症（phenylketonuria,PKU）筛查中的准确性。方法采用茚三酮荧光法对166247例PKU标本进行初筛,初查阳性标本均采用茚三酮荧光法和串联质谱技术复查,复查阳性者召回采血后统一再用上述两种方法进行检测,通过卡方检验统计比较两种方法的复查阳性率,探讨这两种方法在苯丙酮尿症筛查中的应用。根据血中Phe浓度进行鉴别诊断。结果（1）166247例新生儿初查阳性1152例,初查阳性率为6．93‰;（2）茚三酮荧光法和TMs技术的复查阳性率分别为14．58％、3．91％,两种方法比较具有统计学意义（P〈0．05）;（3）复查阳性者召回采血后统一再用上述两种方法进行检测,均确诊HPA3例,发病率为1：55415,其中经典型PKU2例,高苯丙氨酸血症1例。结论TMS技术能更快速、更准确地监测血苯丙氨酸水平,更适合用于PKU筛查,可有效地开展筛查的阳性召回工作及缓解家庭的精神压力。     

Comparison of MB-Check, BACTEC, and egg-based media for recovery of mycobacteria.

The rate of recovery and time to the detection of mycobacteria from clinical specimens were measured for biphasic (MB-Check; Nippon Roche Co., Ltd., Tokyo, Japan) and radiometric (BACTEC; Nippon Becton Dickinson Co., Ltd., Tokyo, Japan) liquid-based culture systems and egg-based media (3% Ogawa and Ogawa K). From the 245 sputum specimens processed, a total of 86 (35.1%) mycobacterial isolates were detected. Of these, 81 (94.2%) and 80 (93.0%) isolates were detected with the MB-Check and BACTEC systems, respectively, and 65 (75.6%) isolates were detected with the 3% Ogawa egg method. The difference in the percentages of positive cultures between the two systems based on liquid media and the 3% Ogawa egg method was significant (P less than 0.01). This difference was even greater among smear-negative specimens. The detection time was shorter with the liquid-based systems. The mean times to the detection of the Mycobacterium tuberculosis complex were 19.1 days with the MB-Check system, 13.4 days with the BACTEC system, and 21.7 days with the 3% Ogawa egg method. These results indicate that both the MB-Check and the BACTEC systems, based on liquid media, are efficient for the recovery of mycobacteria.     

小鼠1-细胞胚胎体外培养条件的研究

目的　检验培养基（Ｗｈｉｔｔｅｎ、ＣＺＢ、ＳＯＭ 和ＫＳＯＭ）、培养器皿（进口、国产新旧平皿）与配制培养基的水质对小鼠胚胎体外发育的影响。　方法　采用微滴培养法培养昆明种小白鼠１－细胞胚胎,以胚胎发育到囊胚的比例为判断培养效果的标准。　结果　Ｗｈｉｔｔｅｎ、ＣＺＢ、ＳＯＭ 和ＫＳＯＭ 培养基中２－细胞发育率分别为１００％ 、１００％ 、１００％ 和９８．６％ ,而囊胚发育率却分别为０％ 、７８．３％ 、４．８％ 和９．５％ ;用三蒸水、五蒸水和去离子水配制的ＣＺＢ培养基,培养囊胚发育率分别为４０．８％ 、５４．１％ 和５２．４％ ;国产和进口新平皿的囊胚发育率分别为６８．０％ 和７２．２％ ;国产和进口旧平皿的培养囊胚发育率为２７．２％ 和４６．１％ 。　结论　昆明小白鼠早期胚胎存在体外发育阻断现象;ＣＺＢ培养基培养昆明小白鼠早期胚胎效果最好;用三蒸水、五蒸水和去离子水配制的ＣＺＢ培养基培养结果无显著差别;进口和国产新平皿的培养效果相近,但再次使用时培养效果显著下降。     

Interference by Neisseria gonorrhoeae growth by other bacterial species.

Growth of Neisseria gonorrhoeae from clinical specimens has been enhanced by the use of selective media that inhibit the simultaneous growth of other microorganisms. One explanation for this enhancement could be that certain other bacteria inhibit gonococcal growth. This hypothesis was examined by testing 167 bacterial isolates for in vitro gonococcal inhibition; 34.1% of the isolates failed to inhibit the gonococcus, but 12.0% produced weak inhibition and 53.9% strongly inhibited N. gonorrhoeae. The pattern of in vitro gonococcal inhibition was consistently the same for all the individual isolates within some species, but individual isolates within other bacterial species varied in their ability to inhibit the gonococcus. Consistently strong in vitro N. gonorrhoeae inhibitors were Citrobacter diversus, Enterobacter cloacae, Serratia marcescens, and Pseudomonas. The in vivo significance of gonococcal interference was demonstrated in the subcutaneous chamber model of N. gonorrhoeae infection.     

Siderophore Production by Pathogenic Neisseria spp

Previous studies have established the importance of iron acquisition to the growth and virulence of Neisseria meningitidis and Neisseria gonorrhoeae. Although preliminary evidence that the Neisseria spp. produce siderophores has been presented, the exact mechanism of iron acquisition has remained obscure. Siderophore production by N. gonorrhoeae and N. meningitidis was induced in two different low-iron media. The iron-reactive siderophores, “gonobactin” and “meningobactin,” were partially purified by ion exchange chromatography followed by extraction with phenol-chloroform-ether or by gel filtration. The compounds were of low molecular weight, their synthesis was repressed by iron in the medium, and they appeared to be hydroxamic acids since they were stimulatory for Arthrobacter flavescens JG-9 (a hydroxamate auxotroph) and gave a positive Csáky reaction for bound hydroxylamine. In the iron form, the compounds had an absorption maximum of approximately 420 nm. Although meningobactin stimulated growth of the gonococcus in low-iron media and vice versa, the homologous activity was more marked, indicating that the compounds, though similar, were probably not identical. As determined by A. flavescens assay the meningococcus produced three to five times more siderophore than did the gonococcus; however, the amount of siderophore present in the culture fluids of even the meningococcus was 100- to 1,000-fold lower than the concentration of hydroxamate siderophores reported to be produced by Bacillus megaterium or Aerobacter aerogenes. Virulent, colony type 1 N. gonorrhoeae produced significantly more gonobactin than did the avirulent colony type 3 gonococci.     

佛山地区841例不孕妇女生殖道感染病原体流行病学调查

目的了解不孕妇女生殖道感染（RTI）的流行情况及其病原体分布。方法对841例不孕妇女的阴道分泌物、宫颈分泌物及血清进行RTI相关病原体的实验室检查。结果841例不孕妇女RTI发生率为59．2％。其中单种病原体感染40．1％，两种及两种以上病原体感染者19．1％。病原体检出率依次为解脲支原体（40．8％）、真菌（24．9％）、沙眼衣原体（15．1％）、人型支原体（7h3％）、滴虫（0．4％）、淋球菌（0．2％）、梅毒（0．2％）、HIV（0％）。结论在不孕妇女中普及RTI相关病原体的筛查，有利于RTI防治及优生优育。     

Comparison of three commercial ELISA kits for the detection of turkey rhinotracheitis virus antibodies.

Three commercial ELISAs (Pathasure, Svanovir and Flockscreen) were compared in their ability to detect turkey rhinotracheitis virus (TRTV) vaccine-induced antibodies. Sequential sera taken from specified pathogen-free chickens, vaccinated with two combinations of live and inactivated TRTV vaccines were used. The vaccines were based on TRTV strains from the United Kingdom (Intervet) and from France (Rhone Merieux). One ELISA failed to detect antibodies after live vaccination with both French and UK vaccines, but detected antibodies induced by both inactivated vaccines by 8 days after vaccination. The two other ELISAs responded to both live vaccinations equally well and both detected a rise in antibody level 8 days after the inactivated vaccination. The specificity of the three ELISAs was 100%. When tested on field samples, all three ELISAs indicated a high prevalence of TRTV antibodies in turkey flocks in the Netherlands.     

Comparative evaluation of AccuProbe culture identification test for Neisseria gonorrhoeae and other rapid methods.

The AccuProbe chemiluminescent culture identification test for Neisseria gonorrhoeae (Gen-Probe Inc., San Diego, Calif.) was assessed in a comparative evaluation with other rapid methods by using 269 isolates of oxidase-positive, gram-negative diplococci. Chemiluminescence was read with a PAL luminometer, and results were expressed as PAL light units (PLUs): the cutoff value for a positive identification was 1,500 PLUs. All 200 isolates of gonococci confirmed by carbohydrate utilization and serotyped with monoclonal antibodies were identified correctly by AccuProbe on initial testing. The API Quadferm system (Bio Merieux, Marcy l'Etoile, France) identified 95% (n = 190) of the gonococci correctly on initial testing and 99.5% (n = 199) on repeat testing, while the Phadebact Monoclonal GC test (Kara Bio Diagnostics AB, Huddinge, Sweden) identified 95.5% (n = 191) of the gonococci on both initial and repeat testing; 8 of the Phadebact-negative isolates were all of the same rare serovar (serovar 1B-17). The mean PLU for the gonococcal isolates was 9,014 (range 2,264 to 10,845) compared with a mean of 51 (range, 8 to 109) for the 69 nongonococcal isolates. We conclude that the AccuProbe culture confirmation test provides a rapid and accurate objective means of identifying cultured N. gonorrhoeae isolates.     

Trimethoprim as an additional selective agent in media for the isolation of N. gonorrhoeae

The incorporation of trimethoprim into existing selective media using either saponin-lysed blood agar or chocolate agar as the base is suggested for the routine isolation of the gonococcus. The principal advantage over other selective media is the inhibition of Proteus mirabilis and other commensal organisms as a consequence of which multiple swabs can be plated out onto a single selective plate irrespective of source, while the recovery rate of the gonococcus from these specimens is increased. The relative merits of both basic media are briefly discussed.     

Changing trends in the antibiograms of Salmonella isolates at a tertiary care hospital in Hyderabad

The present study was undertaken to compare the changing trends of antibiograms of Salmonella enterica serovar Typhi and Salmonella enterica serovar Paratyphi A isolates. A total of 80 isolates of salmonella obtained from blood cultures between 2001-2004 were included in the study. Identification and antibiotic sensitivities of the isolates were performed by using mini API (bio Merieux, France). Sixty isolates were identified as Salmonella enterica serovar Typhi and 20 were identified as Salmonella enterica serovar Paratyphi A. More than 67% of S.typhi and 80% of S.paratyphi A isolates were sensitive to chloramphenicol. Sensitivity of S.typhi isolates to cephalosporins was found to have increased from 2001-2004 while that of S.paratyphi A showed a decline. With increasing resistance to ciprofloxacin and the possibility of re-emergence of sensitivity to chloramphenicol, the policy of empirical treatment of enteric fever needs to be rationalized.     

Study of beta-lactamase in Neisseria gonorrhoeae by a gel adaptation of the iodometric method

A simple protocol for the demonstration of a beta-lactamase in the gonococcus has been adapted from the iodometric technique on agar medium developed by Labia and Barthelemy [13]. This protocol has been applied to 70 strains which produce this enzyme isolated in France and in three African countries.