Ramos免疫组织化学技术的敏感性研究

观察快速微波一步法（Ｒａｍｏｓ法）免疫组织化学技术的敏感性。选用１５种辣根过氧化物酶（ＨＲＰ）标记的特异性抗体（ＥＰＯＳ）,对５９例相应的恶性肿瘤组织进行快速微波免疫组织化学一步法标记。结果显示５８例为阳性（９８．３％）,其中弱阳性４例,中度阳性和强阳性为５４例。结果表明,Ｒａｍｏｓ法具有敏感性高、特异性强、快速而简便等特点,有广阔的应用前景     

EnVision与特异性抗体的一步法免疫组织化学标记

探讨En Vision与特异性抗体复合一步法免疫组织化学标记的可行性及其应用效果。筛选适合于该法的特异性抗体。利用辣根过氧化物酶标记的第二抗体（En Vision）分别与72种单克隆和多克隆特异性抗体混合配制成即用型试剂,将两步标记法变为快速微波一步法,并对2100余例各种良性和亚性肿瘤的免疫组化标记结果进行观察和分析。结果显示绝大发特异性抗体的特异性和敏感性与标准En Vision法相似,其中46种特异性抗体结果稳定,重复性佳;14种特异必抗体稳定性欠佳,但临用前新配制效果仍较理想;12种不理想,不主张用于此法,结果表明En Vision与特异性抗体复合免疫组化一步法是一种有效、快速、简便的免疫组化染色技术。适用于临床病理快速免疫组化诊断,但对所用的特异性抗体应注意选择。     

快速褪黑色素方法在免疫组织化学染色中的应用

目的探讨在恶性黑色素瘤组织中如何快速褪除黑色素,便于进行免疫组织化学检测和观察。方法选取20例恶性黑色素瘤组织石蜡切片,分别应用0.5%高锰酸钾水浴法、0.25%高锰酸钾溶液滴入法、15%过氧化氢浸泡法和传统过氧化氢脱色法进行褪黑色素处理,处理过的切片分别进行免疫组织化学和HE染色,比较染色结果。结果以0.25%高锰酸钾溶液滴入法处理后的效果较为理想:HE染色显示切片完整,色素颗粒完全去除,组织形态和细胞形态清晰;免疫组织化学染色结果清晰易判读,具有较强的敏感性和特异性。结论 0.25%高锰酸钾溶液滴入法可作为恶性黑色素瘤去除色素颗粒的有效方法。     

皮肤组织神经髓鞘几种染色方法的比较

对皮肤组织神经髓鞘应用髓磷脂硷性蛋白myelin basic protein简称MBP）免疫组织化学,劳克坚牢蓝(Luxol fast blue,简称LFB）,砂罗铬花青（solochrome cyanine简称SC）,KOH-HIO4-Schiff四种不同的染色方法进行染色比较,结果显示,MBP法具有特异性强,背景干净,对比清晰等特点。SC、Schiff法髓鞘着色效果亦较好,但背景中其他组织也着色,只要分化适当,LFB法髓鞘着色鲜明,且有一定特异性。     

抗原修复技术在免疫组织化学中的作用

免疫组织化学是一门免疫学与病理形态学相结合的新兴学科 ,是现代生物医学的一项重要方法学 ,具有操作简单、敏感性高、特异性强等特点 ,已经广泛应用于临床病理及其相关学科的研究之中 [1,2 ] 。目前采用甲醛为常规标本固定剂 ,醛基与组织蛋白质中的氨基形成交联使抗原决定簇被遮蔽 ,从而影响免疫组织化学结果 ,而且这种作用随着固定时间增长而加重 [3 ]。抗原修复技术的出现 ,使得原来许多只能在冰冻切片作免疫组织化学得以在石蜡切片上实现 ,抗原修复技术在免疫组织化学中的作用愈加明显 ,并且对免疫组织化学方法的敏感性起着决定性作用[…     

非洲猪瘟抗体胶体金免疫层析检测方法的建立及评价

为建立一种快速、高效、便捷的非洲猪瘟病毒抗体检测方法，本试验采用生物信息学方法筛选得到了性能优异的非洲猪瘟病毒抗原表位多肽P54t，利用P54t多肽作为检测抗体的特异性抗原，建立了一种特异性、敏感性好的胶体金免疫层析抗体检测方法，用于检测待测样品中是否含有非洲猪瘟病毒抗体。结果显示，该方法的敏感性为100%，特异性为100%，最低检测限度可达1:12800，与商品化试剂盒比较，符合率为94.3%。结果表明，该方法敏感性好、特异性强，检测结果准确，可用于非洲猪瘟病毒特异性抗体检测。     

免疫组织化学双标技术中应注意的几个问题

免疫组织化学技术是利用抗原抗体反应进行的检测方法,它具有高度特异性和敏感性。因此：免疫组织化学技术作为一种研究手段在科研和教学工作中得到了较为广泛的应用,而随着各项研究工作的需要,免疫组织化学的双标技术也随之产生且应用越来越广泛。根据我们的实践,免疫组织化学双标技术中应注意的事项如下：（1）固定的时间不宜过长,以免抗原的过度丢失。     

Immuno-PCR法诊断早期梅毒的方法学建立及其应用

目的建立Immuno-PCR法诊断早期梅毒的方法学,评价其灵敏度、特异性、重复性及其临床应用。方法利用基因重组TpN47抗原免疫新西兰兔,制备抗体并用Weston blotting检测;利用抗TpN47抗体作为捕获抗体与血清中TpN47抗原结合,通过链霉亲和素、生物素化抗体、生物素化DNA和PCR扩增等建立Immuno-PCR法检测梅毒螺旋体抗原TpN47体系;评价该方法的灵敏度、特异性和重复性;收集200例临床标本通过Immuno-PCR法、ELISA、TPPA和TURST法进行临床应用比较。结果 Weston blotting结果显示TpN47抗体阳性;Immuno-PCR比ELISA法敏感性强103倍,比TPPA、TURST强105倍;特异性高,重复性好。临床标本中Immuno-PCR法敏感性和特异性分别为86.00%（P〈0.05）和100.00%,ELISA法为71.00%和98.00%,TPPA法为65.00%和100.00%,TRUST法为68.00%和95.00%。结论 Immuno-PCR法检测梅毒螺旋体TpN47抗原敏感性高,特异性强,重复性好,可作为梅毒螺旋体感染的早期诊断方法 。     

催化信号放大系统在原位杂交检测中的应用研究

CSA ( Catalyzed Signal Amplification;催化信号放大 )法首次由 Bobrow等 [1]报道应用免疫酶联和蛋白电泳转移膜的检测 ,1 992年 Adams等[2 ] 将该方法应用于免疫组织化学中 ,Kerstens等 [3 ]又将该方法应用于原位杂交 ( ISH)。应用于免疫组织化学检测较传统的 PAP、ABC、LSAB和 En Vision法相比具有较高的敏感性 ,比 LSAB法要敏感 50 0～1 0 0 0倍 ,较 En Viion法敏感 50～ 1 0 0倍 ,而且稳定性高 ,背景低等特点。本实验室已引进该方法 ,应用于免疫组化和原位分子杂交组化 ,其原理 :杂交后 ,经洗 ,Dig标记探针接生物素化抗 D…     

Reproducibility of morphometric measurements of amyloid after various staining methods

Four histologic staining methods used for detecting amyloid (Congo red, viewed in both normal and polarized light, Sirius red, Crystal violet and Thioflavine T) were applied to heart muscle autopsy samples from 19 patients who suffered from amyloidosis. The amount of amyloid present was evaluated with morphometry (point counting) by five pathologists, and the interobserver reproducibility and variation of point counting in these staining methods were analyzed. The Sirius red method showed the least variation and was the most suitable stain for demonstrating amyloid with respect to reproducibility. Thioflavine T showed the greatest variation and was the least suitable stain with respect to reproducibility. The range of variation was considerable in all staining methods. The results show that stains differ in their specificity and sensitivity in staining amyloid, observers differ in their interpretation of staining results and certain stains result in more uniform interpretations than do others.     

Immunocytochemical detection of estrogen receptors by staining with monoclonal antibodies on cytologic specimens of human breast cancer

A study was undertaken to test the possibility of determining the estrogen receptor (ER) content in human breast cancers by staining with commercial specific monoclonal antibodies (MAbs) on cytologic specimens (touch imprints and fine needle aspirates). The aspirates were suspended in a cell culture medium and cytocentrifuged onto slides to preserve their morphologic characteristics and to allow a proper immunocytochemical staining for ERs. MAb staining for ER was also performed on the respective surgical samples. The staining of cytologic samples for ER showed 100% specificity and 95% sensitivity in comparison to the staining of the histologic samples. Moreover, comparison of the percentage of stained cells in the cytologic specimens to the ER content in the respective surgical specimens, as assayed by the dextran-coated charcoal method, showed the MAb staining of cytologic samples to have 94% specificity and 100% sensitivity. These results support the reliability of MAb staining for ERs in cytologic samples and suggest that it could be the assay of choice in particular clinical settings in the evaluation of primary and recurrent breast cancers.     

Applications of immunocolloids in light microscopy. II. Demonstration of lectin-binding sites in paraffin sections by the use of lectin-gold or glycoprotein-gold complexes

A new procedure is presented for the light microscopic demonstration of specific sugar sequences of oligosaccharides in glycoconjugates by lectins combined with the colloidal gold marker system. Tissue sections from aldehyde-fixed and paraffin embedded rat kidney were stained either in a one-step method with lectin directly bound to particles of colloidal gold or in a two-step method using non-labeled lectin and glycoprotein labeled with colloidal gold. In both methods the presence of lectin-binding sites in the tissue sections is revealed by the appearance of a red coloration that is due to the accumulation of gold particles. The high specificity of the technique is combined with a good sensitivity and resolution as demonstrated by a differential plasma membrane staining in renal epithelial cells. The lectin-gold or glycoprotein-gold complexes remain stable for months and produce a permanent nonbleaching staining.     

Comparison of Histological Techniques to Visualize Iron in Paraffin-embedded Brain Tissue of Patients with Alzheimer’s Disease

Better knowledge of the distribution of iron in the brains of Alzheimer’s disease (AD) patients may facilitate the development of an in vivo magnetic resonance (MR) marker for AD and may cast light on the role of this potentially toxic molecule in the pathogenesis of AD. Several histological iron staining techniques have been used in the past but they have not been systematically tested for sensitivity and specificity. This article compares three histochemical techniques and ferritin immunohistochemistry to visualize iron in paraffin-embedded human AD brain tissue. The specificity of the histochemical techniques was tested by staining sections after iron extraction. Iron was demonstrated in the white matter, in layers IV/V of the frontal neocortex, in iron containing plaques, and in microglia. In our hands, these structures were best visualized using the Meguro iron stain, a method that has not been described for iron staining in human brain or AD in particular. Ferritin immunohistochemistry stained microglia and iron containing plaques similar to the Meguro method but was less intense in myelin-associated iron. The Meguro method is most suitable for identifying iron-positive structures in paraffin-embedded human AD brain tissue.     

Comparison of a nested polymerase chain reaction--restriction fragment length polymorphism method, the PATH antigen detection method, and microscopy for the detection and identification of malaria parasites.

A nested polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method, the PATH antigen detection method, and light microscopy were compared for their capacity to detect and identify Plasmodium species. One hundred and thirty-six blood specimens obtained from patients suspected of having malaria were examined by each of the three methods. Forty-four specimens were positive for malaria using microscopy as the "gold standard". The sensitivity for nested PCR was 100%, and the specificity was 98%. For the detection of Plasmodium falciparum, the antigen detection method had a sensitivity of 100% and a specificity of 97%. Species identification obtained using PCR-RFLP was identical or superior to light microscopy in 42 cases (96%). Although the nested PCR-RFLP method was more sensitive and specific, the rapid turnaround time and high sensitivity of the antigen detection method makes it a useful adjunct to standard microscopy.     

革兰染色对患者阴道分泌物的检验价值

目的探讨革兰染色对患者阴道分泌物的检验价值。方法选择2015年4月至2017年4月于诸暨市中心医院进行阴道分泌物检验的阴道炎患者为研究对象,共1 600例。采用生理盐水镜检法和革兰染色法对分泌物进行检验,比较两种检验方法对分泌物病变的检出情况,同时比较两种检验方法的特异性和敏感性。结果革兰染色法对分泌物病变的检出率为32.50%,生理盐水镜检法的检出率为28.50%,二者差异有统计学意义(P0.05)。革兰染色法对阴道分泌物检验的准确性、敏感性及特异性分别为95.61%、96.34%、97.87%,均高于生理盐水镜检法,差异有统计学意义(P0.05)。结论阴道分泌物检验是诊断阴道炎最便捷的方法。与常规的生理盐水镜检法相比,革兰染色法具有更高的准确性、特异性和敏感性,对阴道炎的早发现、早治疗具有较高的应用价值。     

Comparison of culture, enzyme immunoassay and nucleic acid sandwich hybridization in detecting Chlamydia trachomatis in genital tract infections

Abstract Culture, enzyme immunoassay (Chlamydiazyme™) and nucleic acid sandwich hybridization were compared in detecting Chlamydia trachomatis in uncomplicated genital tract infections. Urethral and cervical specimens were collected from 100 males and 100 females attending a sexually transmitted disease clinic. Chlamydial culture was performed under optimal conditions (duplicate inoculation within the day specimen was collected, culture in vials, monoclonal antibody staining of inclusions, blind passage for negative samples). Here the sensitivity of culture exceeded that of the rapid methods. The sensitivity of a chlamydial antigen detection method (Chlamydiazyme™) was 68% in male and 86% in female specimens, when compared with culture, and the specificity was 100% and 97%, respectively. Acinetobacter calcoace9icus present in clinical specimens did not interfere with Chlamydiazyme™. The sensitivity of the nucleic acid sandwich hybridization was 53% of that of the culture, and specificity 100%. By comparing the three methods it was apparent that the rapid methods did not reveal chlamydial infections not detectable by culture. Thus, if performed carefully, culture is the most sensitive diagnostic method in acute genital infections due to C. trachomatis .     

Criteria of reliability for light microscopic immunocytochemical staining

Summary The following criteria of reliability are defined and discussed for immunocytochemical staining at the light microscopical level: efficiency, accuracy, precision, sensitivity, and specificity. Whenever practical, tests are suggested for obtaining information on the extent to which these criteria are fulfilled in a given system, and procedures are outlined for improving immunocytochemical staining in terms of these criteria. It is suggested that consideration of reliability criteria will help investigators in their choice of methodology, design of experimental strategy, and valid interpretation of the results.     

聚合酶链式反应检测结核杆菌的研究

以人型结核杆菌基因组DNA为模板,合成二段引物各20个碱基进行聚合酶链式反应(PCR)。经琼脂糖凝胶电泳证实,获得一条245bp扩增带。PCR检测的敏感性染色体基因组DNA为1pg,菌悬液为13个活菌／ml。在特异性试验中,人型结核杆趋,牛型结核杆菌、BCG可见此扩增带。被试的其它14种扰酸菌以及变铅青链霉菌、大肠杆菌质粒Puc19、星状诺卡氏菌、红球菌均未见该扩增带。54例肺结核痰标本3种方法检查的阳性率分别为：萋尼氏抗酸染色16．7％,培养法14．8％,PCR 37．0％。前2种检查方法分别与PCR比较,经统计学处理均有显著性差异(P<0．01)。12例非结核性肺部疾患痰标本抗酸染色和PCR均为阴性。结果表明,PCR技术是快速、敏感、特异诊断结核病的方法。     

An evaluation of some complexing methods for the histochemistry of calcium.

A selection of reagents capable of complexing with calcium in various ways was compared by applying them to tissue sections known to contain calcium in a variety of physiochemical states. Their histochemical potential was evaluated according to their sensitivity, specificity, accuracy of localisation and intensity of staining. Murexide, for light microscopy, and 8-hydroxyquinoline, for fluorescence microscopy proved to be the best overall reagents. They failed to demonstrate calcium oxalate, which was well shown by naphthalhydroxamic acid.