三种粪便隐血试验的比较研究

粪便隐血试验是检测消化道微量出血的重要手段。本文用三种隐血试验:血红蛋白定量试验(HQT)、反向间接血球凝集试验(RPHA)和联苯胺试验(BT)对不同消化道疾病患者的粪便进行检测。结果表明隐血的检出用HQT和RPHA较BT更为特异和敏感。     

反向被动血凝试验检测人胃液潜血的探讨

目的：研究一种检测微量胃液潜血的新方法。方法：建立用反向被动血凝试验（ＲＰＨＡ）检测人胃液潜血的方法,检测正常人和胃癌及其癌前疾病等患者共１８５份胃液标本。结果：ＲＰＨＡ法检测血红蛋白（Ｈｂ）的最低浓度为０．６μｇ／ｍｌ,最低检出量为１５ｎｇ。以胃癌和不典型增生阳性率最高（１００％）,其次是萎缩性胃炎（８４．６２％）。正常空腹胃液（ｐＨ１．５）对Ｈｂ的抗原性仅有轻度影响。结论：本ＲＰＨＡ法具有特异性强、敏感性高、方法简便、抗体用量少、无需特殊仪器、结果易观察等优点。适于在胃癌及其癌前疾病的人群筛检中推广应用     

原发性肾小球疾病发病机理探讨——IgE在发病机理中的意义

本文用反向被动血球凝集试验方法检测了23名慢性肾炎肾病型病人及21名正常人的血清IgE水平。21名对照者血清IgE均值为153.815IU/ml,肾炎病人为374.973IU/ml,与对照组相此,差异极显著。治疗后与治疗前相比,11例中有6例血清IgE水平下降,3例IgE上升,2例无改变。结果表明,IgE可能在某些类型的肾小球疾病的发病机理中起一定作用。     

RIA与RPHA两法检测血清HBsAg结果比较研究

以RIA与RPHA两法同时检测了2202例血清标本的HBsAg。RPHA法测得的HBsAg流行率仅为7.99%,而RIA法测得这个率为25.16%。RPHA(对于RIA)的敏感度仅为29.60%,特异度为99.27%。如果采用RPHA结果,我们的肝癌病因学定群研究就显得样本量不足,不能进行分层统计处理。而采用RIA结果,该项定群研究获得了很明确的结论。     

Comparison of the sensitivity of NAT using pooled donor samples for HBV and that of a serologic HBsAg assay

BACKGROUND: Studies were conducted using samples from early and late-stage HBV-infected persons to determine the pool size at which PCR had better sensitivity than a sensitive HBsAg chemoluminescence immunoassay (CLIA-HBsAg). STUDY DESIGN AND METHODS: HBV seroconversion panels were tested for HBsAg by CLIA and for HBV DNA by nested PCR (95% hit rate: 100 copies/mL); PCR was carried out at various dilutions. HBV serologically positive samples that were detected from the simultaneous screening of 540,161 routine whole-blood donations using CLIA-HBsAg and agglutination assays were also characterized for additional markers of HBV infection. RESULTS: In 9 of 10 HBV seroconversion panels, PCR had better sensitivity than CLIA-HBsAg at dilutions of 1-in-25 or lower. Of 65 CLIA-only confirmed-positive donor samples (agglutination assay-negative), 8 represented early infection, 2 of which were PCR positive at a 1-in-50 dilution but negative at a 1-in-100 dilution. Only 2 of 47 samples from probable late-stage HBV infection that were positive on CLIA only were PCR positive with 0.1-mL sample volume and the S-region primer; the remaining 45 samples required a 1.0-mL sample input and C-region primer for increased PCR positivity. The remaining 10 CLIA-only confirmed-positive donor samples were from HBV vaccine recipients. None of the 12 CLIA- and HBsAg-negative donor samples that were strongly anti-HBc reactive could be detected by PCR at any dilution; all 12 were PCR positive when undiluted, but 4 required a 1.0-mL input volume for PCR positivity. CONCLUSION: For the detection of samples representing early-stage HBV infection, PCR at dilutions of 1-in-25 or lower (equivalent to a pool of < or =25 members) had greater sensitivity than CLIA-HBsAg. In contrast, samples from late-stage HBV infection were detected by PCR only with undiluted samples (0.1-mL or 1.0-mL input volumes), regardless of CLIA-HBsAg reactivity. Therefore, although NAT using minipools of 25 donations or less may be effective for the detection of early-stage HBV infection, it may not be effective for the detection of persistent HBV infection.     

点免疫结合试验和反向间接血凝试验检测粪便中福氏志贺氏菌的研究

采用单克隆抗体作为试剂,建立了两种用于快速检测粪便中福氏痢疾杆菌的方法—DIBA夹心法和RPHA法。长期以来困扰研究者的非特异性干扰问题,可能用煮沸标本上清的方法得到解决。检测114份病人标本的结果证明,用常规培养法检测阳性者,DIBA和RPHA检测均为阳性,两类方法的符合率为86.4％。由此证明两种快速免疫学方法不仅能检出完整的细菌细胞,而且也能检出其裂解产物。这两种方法,尤其是RPHA法具有较高的敏感性、特异性和快速性,且不需复杂的仪器设备,加之实验程序简单,甚适于在基层单位及农村地区应用。     

残余牛血清蛋白ELISA检测方法的建立及其初步应用

建立检测疫苗中残余牛血清蛋白(CSP)含量的ELISA检测法。将混合的小牛血清分别免疫鸡和家兔,然后分别提取、纯化鸡卵黄免疫球蛋白(IgY)和兔抗小牛血清抗体IgG。IgY用作包被抗体,IgG用作酶标抗体,四甲基联苯胺(TMB)显色检测残余CSP含量。结果:包被用IgY的最适浓度为25—30μg/mL,用含0.4%明胶的0.01mol/LPBS(pH7.4)溶液作为包被缓冲液,HRPIgG的最适稀释度为1:20001:3000。该方法的检测灵敏度为2.5ng/mL,变异系数为6.3%—9.4%,回收率为90.4%—112.8%,与猪、猴、豚鼠血清皆无交叉反应。10批不含Al(OH)3的疫苗ELISA法与RPHA法(反相间接血凝法)检测结果无显著性差异(P>0.05);而10批含Al(OH)3的疫苗,两种方法检测结果相差很大(P<0.05)。建立的ELISA检测法不受Al(OH)3的影响,简便、特异和灵敏,更适用于检测生物制品中的残余CSP。     

A组轮状病毒一步法RT—PCR检测技术的建立和应用研究

目的 设计A组轮状病毒（RV）的特异性引物,应用Tth酶,建立一步法RT－PCR扩增方案,检测西安地区腹泻幼儿196份粪便标本,并与本室研制建立的反向间接血凝法（RPHA）,聚丙烯酰胺凝胶电泳,市售ELISA试剂盒平行检测对比分析。方法 设计并合成第九基因序列保守区互补的特异性引物,在经典法RT－PCR试验成功的基础上,建立合理的一步法RT－PCR＞要四种检测结果阳性率分别为33．16％,23．47％,21．94％和25％,X^2检验表明,RT－PCR最为敏感。结论 反转录和PCR由Tth酶一步完成,克服了经典法中AMV、RNasin等试剂昂贵,操作繁锁等缺点,很适合临检需要。     

血凝法对肝癌高危人群检测AFP发现早期肝癌的临床研究

本文在肝癌高危人群中应用血凝法(RPHA)检测甲胎蛋白(AFP),在受检38188例中,发现肝癌559例,检出率为1463/10~5,其中早期肝癌55例,检出率为144/10~5。其中有35例小肝癌获手术切除。使我院的肝癌手术切除率从40.4％提高到60.0％。本法具备灵敏、方便、便宜、检出率高等优点,是目前用以发现早期肝癌理想的筛选方法。