滴金免疫测定法（DIGFA）检测血吸虫抗体的研究

目的建立一种简便、快速、敏感、特异的检测血吸虫抗体的新方法。方法用滴金免疫测定法（DIGFA）检测血吸虫病人血清抗体,并以ELISA作平行对照。结果DIGFA和ELISA的敏感性分别为97．1％和95．7％;特异性分别为99．1％和100％,两法无显著差异（P＞0．05）。62份其它寄生虫感染血清,除肺吸虫1例,ELISA呈阳性反应外,其余均为阴性。结论实验结果不仅显示了DIGFA的敏感性和特异性与ELISA相似,而且具有简便、快速、不需要特殊设备等优点,是一种值得推广应用的检测血吸虫抗体的新方法。     

应用斑点金免疫渗滤法诊断日本血吸虫病

目的：探索一种便捷的血吸虫病诊断方法。方法：在原已建立的ELISA法的基础上，应用3个针对日本血吸虫抗原的不同表位的单克隆抗体标记胶体金，建立检测日本血吸虫病患者血清中血吸虫抗原的斑点金免疫渗滤法。抗原与抗体通过渗滤在硝酸纤维薄膜上进行反应，数分钟用肉眼观察结果。结果：本法检测SEA的敏感度为16ng／ml，在69例慢性日本血吸虫病患者血清的检测中，阳性率为60．8％，特异性为95．2％；同时用dot－ELISA检测137例慢性日本血吸虫病患者，阳性率为54．7％、特异性为94．6％；夹心ELISA检测118例 慢性日本血吸虫病患者，阳性率为61．9％、特异性为95．7％。结论：经数理统计G检验证实斑点金免疫渗滤法的敏感性和特异性同ELISA 相近, 且操作更简便且快速。     

McAbDot-ELISA检测卫氏并殖吸虫循环免疫复合物

应用Ｄｏｔ－ＥＬＩＳＡ法，以抗卫氏并殖吸虫囊蚴单克隆抗体ＩＦ８Ａ３检测了卫氏并殖吸虫感染犬血清循环免疫复合物动态。感染后１～２ｗｋ可以检测到免疫复合物，３～４ｗｋ阳性滴度逐渐上升，５～６ｗｋ处于较高水平，第８ｗｋ达高峰，然后较快地下降。研究结果表明，卫氏并殖吸虫囊蚴抗原特异的循环免疫复合物具有明显的期特异性；单克隆抗体Ｄｏｔ－ＥＬＩＳＡ检测特异的循环免疫复合物，可用于卫氏并殖吸虫病的早期诊断和活动性感染的诊断。     

金标免疫渗滤法诊断试剂盒诊断血吸虫病的价值研究

目的　评价金标免疫渗滤法诊断试剂盒（DIGFA-kit）用于血吸虫病血清流行病学调查的价值。方法　采用DIGFA与ELISA单盲平行检测各期血吸虫病病人血清、非流行区人群血清、疫区人群血清和血吸虫病已控制地区人群血清共2304份。结果　DIGFA检测血吸虫抗体的敏感性为96.8%，对非流行区健康人群的特异性为100%；该法检测疫区居民血清508份，抗体阳性检出率为35.6%，在Kato法粪检阳性的81人中，其中抗体阳性79人，阳性符合率为97.5%；检测血吸虫病已控制地区人群血清，抗体阳性率为4.6%；根据DIGFA的敏感性和对非流行区人群的特异性，计算其Youdens指数为0.926。上述各项结果与ELISA比较，差异无显著性（P>0.05）。结论　DIGFA与ELISA有相似的敏感性和特异性，且方法简便、快速，胶体金标记物制备容易，试剂稳定，运送方便，在现场和临床诊断更具有优越性。     

滴金免疫测定法检测抗体在血吸虫病传播阻断地区的应用

目的探讨滴金免疫测定法检测抗体在血吸虫病传播阻断地区的应用价值。方法在已建立的滴金免疫测定法的基础上,应用该法检测原血吸虫病重度流行区、中度流行区和轻度流行区而现已阻断传播地区的人群以及现血吸虫病中度流行区人群的抗血吸虫抗体。结果原血吸虫病重、中和轻度流行区人群的阳性检出率分别为６．１８％、３．３１％、１．４７％,三者间有非常显著差异（Ｐ＜０．０１）;现血吸虫病中度流行区人群的阳性检出率为 ３９．３３％,与传播阻断地区人群的检出率间也有非常显著差异（Ｐ＜０．０１）。结论摘金免疫测定法检测抗体可作为考核该类地区血吸虫病防治效果的一种方法。     

Tuberculosis control in Kawasaki City--promoting the DOT program

Kawasaki City initiated the DOT (Directly Observed Therapy) program in 2000 to improve tuberculosis (TB) control in the homeless persons and individuals living in temporary subsidized housing. The program was implemented as an additional program to the city's control policy of mass TB screenings for these groups. The DOT program was implemented through participation of numerous agencies sharing common goals. Through coordination and cooperation, a strategy suitable for the regional level was developed. Sectors involved were not only the TB control staff within the public health centers but also the entire public health centers, the welfare administrative division, the city's TB task force and homeless task force, and regional medical institutions. With the implementation of the DOT program, proper medical treatment, improved patient care, and various other measures were performed, and this led to the strengthening of TB control not only in designated DOT-implemented areas but also in the entire city. As a result, the treatment outcomes improved, and incidence rate of TB began to decrease.     

TB treatment: as simple as DOT?

Treatment completion continues to be an unmet challenge in tuberculosis (TB) control worldwide. Since the introduction of DOTS by the World Health Organization in 1993, directly observed therapy (DOT) has been promoted as the centerpiece of the strategy. The results of three randomized controlled studies designed to evaluate the effects of DOT on treatment completion, and carried out in South Africa, Thailand and Pakistan, showed conflicting results. It is doubtful that any of the studies provided valid measures of the effectiveness of DOT. Each study operationalized DOT in ways that did not match the study definition and failed to account for additional study interventions in their analyses. Given the evidence, there is no one factor that is 'the' answer to the challenge of TB treatment completion.     

酶免疫斑点试验作钩体病血清学诊断的研究

本文用优选的最适条件对619份钩体病人血清和520份对照血清进行IgM-EIDT和MAT检测比较,发现早期病人血清IgM-EIDT阳性率显著高于MAT(P<0.001),恢复期血清则无差异。IgM-EIDT的诊断敏感性为95.53％,特异性为96.73％,两种方法对194份钩体病人双向血清的滴度检测呈显著正相关(r=0.7670)。对部分钩体病人血清用IgG-EIDT检测,其阳性率与MAT相似。因此,IgM-EIDT和IgG-EIDT结合使用,既可作钩体病的早期诊断,也可作追溯诊断和血清流行病学调查。     

Practice of directly observed treatment (DOT) for tuberculosis in southern Thailand: comparison between different types of DOT observers.

SETTING: A government health system in southern Thailand where the directly observed treatment, short-course (DOTS) strategy has been implemented. OBJECTIVE: To compare the practice of actual directly observed treatment (DOT) and the observer sustainability for different types of observer. METHODS: During 1999-2000, 411 patients with new smear-positive pulmonary tuberculosis were followed up. The patients and/or their observers were interviewed about the presence of any person with the patient during drug intake and the practice of watching the patient swallowing the medicine (actual DOT). Data were recorded monthly and analysed by Cox and logistic regression models. RESULTS: For health personnel (HP), community member (CM), and family member (FM) observers, the proportions who did not practise actual DOT were respectively 11%, 23%, and 35%, and the proportions who changed to no observer or self administration were respectively 11%, 1%, and 2%, during the first 9 months of treatment. Health personnel had the lowest risk of not practising actual DOT (odds ratio HP/FM 0.1, 95%CI 0.1-0.2; CM/FM 0.9, 95%CI 0.5-1.6) but the highest risk for change to self administration. CONCLUSION: To increase the coverage of actual DOT, strategies are needed to maintain health personnel as the DOT observers and to promote actual DOT among family member observers.     

Doubts about DOT: antiretroviral therapy for resource-poor countries

INTRODUCTION: Directly observed therapy programs developed for tuberculosis (TB) have been suggested as a model for the provision of HIV medications in resource-poor countries in order to ensure adherence and prevent drug resistance. METHODS: Opinions were formed based on a review of scientific literature regarding the effectiveness of witnessed dosing in directly observed TB therapy programs, adherence to HIV antiretroviral therapy in resource-rich and resource-poor settings, relationship between adherence and HIV antiretroviral drug resistance, HIV viral load and risk of HIV transmission, and stigmatization concerns related to HIV and TB in resource-poor settings. RESULTS/CONCLUSIONS: We suggest that the enthusiasm for HIV directly observed therapy programs is premature based on: equivocal evidence that witnessed dosing is superior to self administered therapy; mistaken assumptions that resource-poor countries are a 'special case' with respect to adherence; possible paradoxical impact of good adherence on HIV drug resistance; unproven efficacy of antiretroviral therapy in preventing HIV transmission; and potential stigmatization of daily antiretroviral dosing.     

问号赖型钩端螺旋体重组质粒pDL121的初步研究

目的　探讨问号赖型钩端螺旋体抗原基因的特点。方法　应用６ 种常见内切酶（ＢａｍＨＩ,Ｂｇｌ Ⅱ,ＥｃｏＲⅠ,Ｈｉｎｄ Ⅲ,ＰｓｔⅠ,Ｐｖｕ Ⅱ）对从问号赖型钩端螺旋体０１７ 株的基因文库中筛选出来的重组质粒ｐＤＬ１２１ 外源基因进行酶谱分析,同时用Ｄｉｇｏｘｉｎ 标记的ｐＤＬ１２１ 外源基因片段作探针对不同种属的致病性钩端螺旋体和非致病性钩端螺旋体基因进行杂交分析。结果　显示问号赖型钩端螺旋体重组质粒ｐＤＬ１２１ 外源基因没有６ 种内切酶的酶切位点,重组探针与致病性钩体（ｓｅｒｏｖａｒｌａｉｓｔｒａｉｎ０１７,ｓｅｒｏｖａｒ ｈｅｂｄｏｍａｄｉｓｓｔｒａｉｎ ５６６１０ ,ｓｅｒｏｖａｒ ｐｏｍｏｎａｓｔｒａｉｎ ５６６０８）有杂交信号,与非致病性钩体（ｓｅｒｏｖａｒ ｐａｔｏｃ ｓｔｒａｉｎ Ｐａｔｏｃ Ⅰ,ｓｅｒｏｖａｒｉｌｌｉｎｉｓｔｒａｉｎ ３０５５）无杂交信号,亦不识别大肠杆菌。结论　重组质粒ｐＤＬ１２１ 外源基因可能是问号赖型钩端螺旋体的一个新基因,进一步测序分析将揭示该基因本质,同时因该重组探针能鉴别致病性钩体和非致病性钩体,提示其可用于钩端螺旋体病的早期诊断及钩端螺旋体的鉴定和分类。     

采用单引物AP-PCR技术对胃癌相关基因片段的分离分析

目的 分离与胃癌发生相关的基因片段并予以鉴定。方法     

贫血患者血清转铁蛋白受体测定的临床意义

采用Ｄｏｔ－ＥＬＩＳＡ法定量测定了６８例不同贫血患者的血清转铁蛋白受体（ｓＴＩＲ）水平,发现缺铁性贫血,溶血性贫血和急性失血性贫血患者ｓＴＩＲ均显高于正常人（Ｐ〈０．００１）,而慢性再生障碍性贫血患者ｓＴｆＲ显著低于正常人（Ｐ〈０．００１）,缺铁性贫血ｓＴｆＲ水平与血清铁蛋白水平呈负相关（Ｐ〈０．００５）,溶血性贫血ｓＴｆＲ与网织红细胞计数呈正相关（Ｐ〈０．００５）,ｓＴｆＲ可以反映机体贮存铁和骨     

斑点金-薄层色谱法检测鼠疫耶尔森氏菌的实验研究

目的建立一种简便、快速的斑点金-薄层色谱法(DIGFA-TLC)用于鼠疫耶尔森氏菌的快速定量诊断。方法用柠檬酸钠还原法制备10 nm的胶体金标记鼠疫耶尔森氏菌多价抗体,制备成鼠疫抗体金探针,通过薄层色谱扫描对样品斑点进行定量测定。结果斑点金-薄层色谱法对鼠疫FI抗原的最低检出限为5 ng;对EV76的最低检出限为1.2×105cfu;且二者呈高度的正相关关系,前者相关直线方程为y=448.467 37+2.511 43x,后者相关直线方程为y=375.760 88+4.112 31×10-4x;整个反应过程在10 min内完成。结论该方法简便、快速、特异、敏感,可以实现鼠疫耶尔森氏菌的定量监测。     

斑点金免疫渗滤法快速诊断布鲁杆菌病的研究

目的建立一种布鲁杆菌病快速免疫诊断方法。方法以硝酸纤维素膜为载体,胶体金标记蛋白为显示剂,通过最佳实验条件的选择及稳定性、重现性、敏感性、特异性和交叉反应试验,建立斑点金免疫渗滤法(DIGFA)快速检测布鲁杆菌病。结果 DIGFA快速检测布鲁杆菌病具有很好的重现性和稳定性;敏感性、特异性分别达到98％和100％;未发现与肺结核患者、乙肝患者及幽门螺杆菌感染者血清有交叉反应。结论 DIGFA快速检测布鲁杆菌病快速、简便,测试过程1-2 min内完成,试剂敏感、特异、稳定,非常适合布鲁杆菌病的诊断及流行病学调查。     

血吸虫病患者血清中循环25kD表膜抗原的检测

采用细胞杂交瘤技术获得一株血吸虫成虫抗原的单克隆抗体SM 38-3。其靶抗原为成虫表膜,免疫球蛋白型为1gG 2a,能识别血吸虫分子量为25k D的表膜蛋白抗原表位。该单克隆抗体纯化后标记过氧化物酶,用于Dot-ELISA测定慢性血吸虫病人血清115份,阳性反应103份,阳性率为89.6％;测定急性病人血清89份,阳性反应85份,阳性率为95.5％;测定141份正常人血清,阳性反应2份,假阳性率为1.4％。与血吸虫肠相关抗原的检测比较,结果阳性率与抗原滴度均无明显差别。