Fluorescent Antibody Technique, Direct

荧光抗体技术,直接
  • 文章类型: Case Reports
    背景:慢性光敏性皮炎(CPD)(也称为光化性网状蛋白)是一种罕见的疾病,通常在老年男性中提及。受影响的患者严重瘙痒,多年来暴露在阳光下的干性皮肤变厚。
    方法:一名高加索女性患者,她的大部分时间都在外面工作,脖子上有“慢性皮炎”,她在阳光明媚的日子开始在她的花园里种植菊花。稍后,她出现了水肿,红斑,丘疹,脖子上有几个囊泡严重瘙痒.
    方法:皮肤活检显示诊断为CPD,随着紫外线B(UVB)的积极测试,UVB(MEDB)UVA(MEDA)和PhotoPath的最小红斑剂量(MED)。
    结果:使用抗人纤维蛋白原抗体的直接免疫荧光(DIF)染色,白蛋白,IgG,IgM,lambda,kappa,真皮表皮连接处基膜区C3c和C1q呈阳性,在乳头状真皮中,以及所有真皮和细胞外基质中的神经血管束,尤其是那些在水泡下的。
    结论:在这种情况下,我们建议不要忘记在重新激活的CPD病例中使用DIF以及照片补丁测试的重要性.
    Chronic photosensitivity dermatitis (CPD) (also named actinic reticuloid) is an unusual disease classically referred often in elderly men. Affected patients have severely itchy, thickened dry skin in areas exposed to the sun throughout the years.
    A Caucasian female patient who worked most of her life outside who had \"chronic dermatitis\" in her neck started planting chrysanthemum in her garden on a sunny day. Later, she presented edema, erythema, papules, and a few vesicles in her neck with severe pruritus.
    A skin biopsy revealed the diagnosis of CPD, along with positive testing for ultraviolet B (UVB), minimal erythema doses (MED) for UVB (MEDB) UVA (MEDA) and PhotoPath.
    Direct immunofluorescence (DIF) stains using anti-human antibodies against fibrinogen, albumin, IgG, IgM, lambda, kappa, and C3c and C1q were positive at the base membrane area of the dermal epidermal junction, in the papillary dermis, as well as the neurovascular bundles in all the dermis and the extracellular matrix, especially those under the blisters.
    With this case, we suggest not forgetting the importance of using DIF in reactivated CPD cases in addition to the photo patch testing.
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  • 文章类型: Letter
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  • 文章类型: Journal Article
    十二指肠贾第鞭毛虫(syn。G.intelinalis或G.lamblia)是一种寄生原生动物,可感染多种宿主的上肠道,包括人类和家畜。因此,它引起了人们对伴侣动物造成的公共卫生风险的担忧。最近,随着生活水平的提高和宠物与人类接触的增加,贾第虫的人畜共患传播急剧增加。从基因的角度来看,十二指肠G.应该被视为一个复杂的物种,包括八个不同的物种特异性遗传组合。实验室诊断主要基于通过联合检查在粪便样本中发现微小囊肿。其他方法包括检测抗原,免疫测定或PCR方案,可以识别贾第虫组合。该研究旨在比较直接荧光抗体测试(DFA)的性能,硫酸锌浮选技术(ZnSO4),快速诊断测试(RDT),终点PCR扩增(PCR)用于检测贾第鞭毛虫并鉴定来自意大利中部的犬种群中的相关组合。直接荧光抗体测试是实验室诊断犬粪便样本中贾第虫的参考标准,尽管浮选后的显微镜检查仍然是许多兽医诊断中心最有用的方法。本发现证明了DFA和ZnSO4在检测贾第虫中的高性能,而RDT可用作DFA和ZnSO4的替代或补充方法。PCR性能较低,但它允许在25%的PCR阳性标本中确定贾第虫B人畜共患病组合(60个中的15个),而其余的PCR阳性分离株属于狗特异性组合C。DFA在自己的狗中检测到26%的十二指肠G。以及潜在的人畜共患组合的鉴定,突显了公共健康的潜在风险,并表明寄生虫在人和狗之间频繁的跨物种传播。
    Giardia duodenalis (syn. G. intestinalis or G. lamblia) is a parasitic protozoan that infects the upper intestinal tract of a broad range of hosts, including humans and domestic animals. Thus, it has raised concerns about the public health risk due to companion animals. Recently, with the improvement of living standards and increasing contacts between pets and humans, the zoonotic transmission of Giardia has dramatically increased. From a genetic point of view, G. duodenalis should be viewed as a complex species that includes eight different species-specific genetic assemblages. The laboratory diagnosis is mainly based on the finding of microscopic cysts in stool samples by coprological examination. Other methods include the detection of antigens, immunoassays or PCR protocols, which allow the identification of Giardia assemblages. The study aimed to compare the performance of Direct Fluorescence Antibody test (DFA), zinc sulfate flotation technique (ZnSO4), rapid diagnostic test (RDT), end-point PCR amplification (PCR) for the detection of Giardia and to identify the concerning assemblages in a canine population from Central Italy. Direct fluorescence antibody test is the reference standard for laboratory diagnosis of Giardia in fecal samples from dogs, despite the microscopic examination after flotation remains the most useful method in many veterinary diagnostic centers. The present findings demonstrate the high performance of DFA and ZnSO4 in detecting Giardia, while RDT may be useful as alternative or complementary method to the DFA and ZnSO4. PCR performance was low, but it allowed determining Giardia B zoonotic assemblage in 25% of the PCR-positive specimens (15 out of 60), while the remaining PCR-positive isolates belonged to the dog-specific assemblage C. The 26% prevalence of G. duodenalis detected by DFA in owned dogs and the identification of potentially zoonotic assemblages underline the potential risk for public health and indicate frequent cross-species transmission of the parasite between humans and dogs.
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  • 文章类型: Journal Article
    我们对Tifton兽医诊断和研究实验室(Tifton,GA,美国)2010年7月至2021年6月。提交了来自佐治亚州89个县的23种动物的792个样本,和佛罗里达州的4个邻近县,1在南卡罗来纳州,1在阿拉巴马在13例(1.6%)中,DFAT结果尚无定论;779例(98.4%)病例的检测结果为结论性(阳性或阴性).在这779个案例中,79(10.1%)在10个物种中测试为阳性。其余700例(89.9%)为阴性。提交RABV测试的主要原因是在414例(52.3%)病例中人类暴露于潜在的狂犬病动物。在79例阳性病例中,74例(93.7%)涉及野生动物;浣熊(51例;68.9%)是确诊为RABV感染的主要宿主,其次是臭鼬和狐狸(各8例;10.8%),山猫(5例;6.8%),和蝙蝠(2例;2.7%)。在我们的研究期间,只有5只家畜(阳性病例的6.3%)检测呈阳性;每个牛,犬,caprine,马,和猫科动物。因此,在我们的研究区域中,循环循环在循环RABV感染中起主要作用。
    We performed a retrospective study of all case submissions for the rabies virus (RABV) direct fluorescent antibody test (DFAT) requested of the Tifton Veterinary Diagnostic and Investigational Laboratory (Tifton, GA, USA) between July 2010 and June 2021. Submitted were 792 samples from 23 animal species from 89 counties in Georgia, and 4 neighboring counties in Florida, 1 in South Carolina, and 1 in Alabama. In 13 (1.6%) cases, the DFAT result was inconclusive; 779 (98.4%) cases had a conclusive (positive or negative) test result. Of these 779 cases, 79 (10.1%) tested positive across 10 species. The remaining 700 (89.9%) cases were negative. The main reason for submission for RABV testing was human exposure to a potentially rabid animal in 414 (52.3%) cases. Among the 79 positive cases, 74 (93.7%) involved wildlife; raccoons (51 cases; 68.9%) were the primary host confirmed with RABV infection, followed by skunk and fox (8 cases each; 10.8%), bobcat (5 cases; 6.8%), and bats (2 cases; 2.7%). Only 5 domestic animals (6.3% of the positive cases) tested positive during our study period; one from each of the bovine, canine, caprine, equine, and feline species. Hence, the sylvatic cycle plays the predominant role in circulating RABV infection in our study area.
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  • 文章类型: Journal Article
    目的:本研究的目的是报道年轻患者眼瘢痕性类天疱疮(OCP)的发生。还回顾了相关文献。
    方法:回顾了2021年8月至2023年5月在三级护理眼科研究所接受OCP治疗的30岁或以下患者的医疗记录。瘢痕性结膜炎最常见的鉴别诊断,比如史蒂文斯-约翰逊综合征后遗症,化学损伤,慢性局部/全身用药,自身免疫性结缔组织疾病,或者过敏性眼病,根据临床病史排除,考试,和Shanbhag等人描述的愈合性结膜炎评分。大多数患者的口腔粘膜和/或结膜活检的直接免疫荧光阳性证实了OCP的诊断。
    结果:7例患者符合OCP诊断标准。演示时的平均年龄为21.86±5.25岁(13-28岁)。一些患者表现为OCP的相对非典型特征,例如角膜免疫环浸润和双侧角膜穿孔。6例患者出现全身粘膜病变,直接免疫荧光产率为85.71%。所有患者都需要积极的免疫抑制治疗。
    结论:OCP通常被描述为一种罕见的疾病,发生在老年。此病例系列强调了在年轻时诊断OCP的更高怀疑指数的重要性。早期施用免疫抑制剂可以潜在地控制严重的眼表炎症及其后遗症。
    OBJECTIVE: The purpose of this study is to report ocular cicatricial pemphigoid (OCP) occurring in young patients. Relevant literature is also reviewed.
    METHODS: Medical records of patients aged 30 years or younger who were treated for OCP between August 2021 and May 2023 at a tertiary care eye institute were reviewed. The most common differential diagnoses of cicatrizing conjunctivitis, such as Stevens-Johnson syndrome sequelae, chemical injury, chronic topical/systemic drug use, autoimmune connective tissue disorders, or allergic eye diseases, were ruled out based on clinical history, examination, and the Cicatrizing Conjunctivitis Score described by Shanbhag et al. The diagnosis of OCP was confirmed by positive direct immunofluorescence of oral mucosal and/or conjunctival biopsy in a majority of the patients.
    RESULTS: Seven patients fulfilled the criteria for a diagnosis of OCP. The mean age at presentation was 21.86 ± 5.25 years (13-28 years). Some of the patients presented with relatively atypical features for OCP such as corneal immune ring infiltrate and bilateral corneal perforation. Six patients exhibited systemic mucosal lesions, and the direct immunofluorescence yield was 85.71%. All patients required aggressive immunosuppressive treatment.
    CONCLUSIONS: OCP is classically described as a rare disease that occurs in old age. This case series highlights the importance of a higher index of suspicion for diagnosing OCP at a younger age. Early administration of immunosuppressive agents can potentially control severe ocular surface inflammation and its sequelae.
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  • 文章类型: Journal Article
    为了确定临床,组织病理学和免疫荧光作为表皮内免疫性疾病的诊断方法。
    方法:前瞻性横断面研究在皮肤病研究所进行,卡拉奇,从2020年12月到2022年12月,包括任何性别的成年患者,囊泡,皮肤或粘膜上的脓疱和结皮。通过临床评估获得的每位患者的诊断结果,显微镜和直接免疫荧光进行了比较。数据采用SPSS19进行分析。
    结果:在81名患者中,男性41人(50.6%),女性40人(49.4%)。总体中位年龄为35岁(四分位距:23岁),66(75%)患者年龄19-55岁。涉及的主要身体部位是躯干49(60.5%),其次是粘膜26(32.1%)。临床诊断80例(98.7%),相比之下,76(93.8%)通过显微镜和81(100%)通过直接免疫荧光。
    发现直接免疫荧光是确认性诊断表皮内免疫性疾病的金标准,尤其是当临床和组织病理学结果没有定论时。
    UNASSIGNED: To determine the concordance among clinical, histopathological and immunofluorescence as diagnostic methods for intraepidermal immunobullous disorders.
    METHODS: The prospective cross-sectional study was conducted at the Institute of Skin Diseases, Karachi, from December 2020 to December 2022, and comprised adult patients of either gender presenting with complaints of bullae, vesicles, pustules and crusts on the skin or mucous membrane. Diagnostic findings of each patient as obtained by clinical assessment, microscopy and direct immunofluorescence were compared. Data was analysed using SPSS 19.
    RESULTS: Of the 81 patients, 41(50.6%) were males and 40(49.4%) were females. The overall median age was 35 years (interquartile range: 23 years), with 66(75%) patients aged 19-55 years. The predominant body site involved was the trunk 49(60.5%), followed by mucosa 26(32.1%). Clinical diagnosis detected 80(98.7%) cases, compared to 76(93.8%) by microscopy and 81(100%) by direct immunofluorescence.
    UNASSIGNED: Direct immunofluorescence was found to be the gold standard for a confirmatory diagnosis of intraepidermal immunobullous disorders, especially when clinical and histopathology findings were inconclusive.
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  • 文章类型: Journal Article
    背景:人工智能(AI)正在重塑医疗保健,使用机器和深度学习来加强疾病管理。皮肤科已经改善了诊断,特别是在皮肤癌检测中,通过整合AI。然而,AI在自身免疫性大疱性皮肤病的免疫荧光成像自动化方面的潜力仍未开发.虽然直接免疫荧光(DIF)支持诊断,它的人工解释会妨碍效率。使用深度学习对DIF模式进行自动分类,包括细胞间模式(ICP)和线性模式(LP),有望改善自身免疫性大疱性皮肤病的诊断。
    目的:本研究的目的是开发用于自身免疫性大疱性皮肤病DIF模式自动分类的AI算法,如ICP和LP。这旨在提高诊断准确性,简化疾病管理,并通过深度学习驱动的免疫荧光解释改善患者的预后。
    方法:我们收集了2022年1月至2024年1月期间疑似AIBD患者皮肤活检的免疫荧光图像。通过5mm穿刺活检获得皮肤组织,为直接免疫荧光准备。经验丰富的皮肤科医生将图像分为三类:ICP,LP,和消极。为了评估我们的深度学习方法,我们将图像分为训练集(436)和测试集(93)。我们采用了预训练深度神经网络的迁移学习,并进行了5倍交叉验证来评估模型性能。我们的数据集的类不平衡是使用加权损失和数据增加策略解决的。使用Pytorch对模型进行了50个时期的训练,为CNN和SwinTransformer实现224x224的图像大小。
    结果:我们的研究比较了六个CNN和用于AIBDs图像分类的Swin变换器,Swin变压器实现了98.5%的最高平均验证精度。在单独的测试集上,最佳模型达到了94.6%的准确率,在AIBDs类别中表现出95.3%的敏感性和97.5%的特异性。使用Grad-CAM进行的可视化突出了模型对特征模式的依赖,以实现准确的分类。
    结论:该研究强调了CNN在识别DIF特征方面的准确性。这种方法有助于自动分析和报告,提供再现性,速度,数据处理,和成本效益。在皮肤免疫荧光中集成深度学习有望在皮肤科的这一分支中进行精确的诊断和简化的报告。
    BACKGROUND: Artificial intelligence (AI) is reshaping healthcare, using machine and deep learning (DL) to enhance disease management. Dermatology has seen improved diagnostics, particularly in skin cancer detection, through the integration of AI. However, the potential of AI in automating immunofluorescence imaging for autoimmune bullous skin diseases (AIBDs) remains untapped. While direct immunofluorescence (DIF) supports diagnosis, its manual interpretation can hinder efficiency. The use of DL to classify DIF patterns automatically, including the intercellular (ICP) and linear pattern (LP), holds promise for improving the diagnosis of AIBDs.
    OBJECTIVE: To develop AI algorithms for automated classification of AIBD DIF patterns, such as ICP and LP, in order to enhance diagnostic accuracy, streamline disease management and improve patient outcomes through DL-driven immunofluorescence interpretation.
    METHODS: We collected immunofluorescence images from skin biopsies of patients suspected of having an AIBD between January 2022 and January 2024. Skin tissue was obtained via a 5-mm punch biopsy, prepared for DIF. Experienced dermatologists classified the images as ICP, LP or negative. To evaluate our DL approach, we divided the images into training (n = 436) and test sets (n = 93). We employed transfer learning with pretrained deep neural networks and conducted fivefold cross-validation to assess model performance. Our dataset\'s class imbalance was addressed using weighted loss and data augmentation strategies. The models were trained for 50 epochs using Pytorch, achieving an image size of 224 × 224 pixels for both convolutional neural networks (CNNs) and the Swin Transformer.
    RESULTS: Our study compared six CNNs and the Swin Transformer for AIBD image classification, with the Swin Transformer achieving the highest average validation accuracy (98.5%). On a separate test set, the best model attained an accuracy of 94.6%, demonstrating 95.3% sensitivity and 97.5% specificity across AIBD classes. Visualization with Grad-CAM (class activation mapping) highlighted the model\'s reliance on characteristic patterns for accurate classification.
    CONCLUSIONS: The study highlighted the accuracy of CNNs in identifying DIF features. This approach aids automated analysis and reporting, offering reproducibility, speed, data handling and cost-efficiency. Integrating DL into skin immunofluorescence promises precise diagnostics and streamlined reporting in this branch of dermatology.
    Artificial intelligence (AI) is transforming healthcare through machine and deep learning (computer systems that can learn and adapt, and make complex decisions, without receiving explicit instructions), improving disease management in dermatology, particularly in detecting skin cancer. However, AI’s potential in automating immunofluorescence imaging in autoimmune bullous (blistering) skin diseases (AIBDs) remains largely untapped. Manual interpretation of direct immunofluorescence (DIF – a type of microscopy) can reduce efficiency. However, using deep learning to automatically classify DIF patterns (for example, the ‘intercellular pattern’ (ICP) and the ‘linear pattern’ (LP)) holds promise in helping with the diagnosis of AIBDs. This study aimed to develop AI algorithms for the automated classification of AIBD DIF patterns, such as ICP and LP, to improve diagnostic accuracy and streamline disease management. Immunofluorescence images were collected from skin biopsies of patients with a suspected AIBD between January 2022 and January 2024. Dermatologists classified the images into three categories: ICP, LP and negative. The dataset was divided into training (436 images) and test sets (93 images). A transfer learning framework (where what has been learned previously in one setting is used to improve performance in another) was used to make up for the limited amount of training data, to explore different models for the AIBD classification task. Our results revealed that a model called the ‘Swin Transformer’ achieved an average accuracy of 99% in diagnosing different AIBDs. The best model attained 95% accuracy on the test set and was reliable in identifying and ruling out different AIBDs. Visualization with Grad-CAM (a technique used in deep learning) highlighted the model’s use of characteristic patterns to classify the diseases accurately. Overall, integrating deep learning in skin immunofluorescence promises to improve diagnostics and streamline reporting in dermatology, which could improve consistency, speed and cost-efficiency.
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  • 文章类型: Journal Article
    大疱性类天疱疮(BP),一种自身免疫性表皮下起泡疾病,显示与荨麻疹红斑相关的紧张水泡。通过直接免疫荧光(DIF)检测到的基底膜区(BMZ)处的组织结合免疫球蛋白G(IgG)是诊断BP的有力证据。补体成分3(C3)的DIF敏感性高于IgG,但是这种不同敏感性的原因还没有完全理解。在这项研究中,我们进行了几项离体研究,以研究在某些BP病例中,通过DIF在BMZ出现IgG阴性和C3阳性的可能机制.首先,发现通过DIF显示IgG阴性的BP患者的血清在他们自己的DIF皮肤样本中明显与BMZ反应。接下来,使用用不同pH的磷酸盐缓冲盐水(PBS)稀释的血清进行间接免疫荧光(IIF),pH7.4、6.0和3.0。用pH7.4PBS稀释的患者血清在BMZ显示线性染色,但用pH6.0PBS和pH3.0PBS稀释的血清显示较低的荧光强度。最后,将BP患者的皮肤切片用不同pH的PBS(pH3.0、6.0和7.4)预孵育,然后用抗人IgG和C3染色。已经用pH3.0PBS和pH6.0PBS预温育的IgG和C3的荧光强度显著低于已经用pH7.4PBS预温育的IgG和C3的荧光强度。这些结果表明,低pH条件阻碍了自身抗体与BMZ的结合,也就是说,炎症诱导的组织pH下降抑制了自身抗体在BMZ的沉积。此外,组织pH值的下降导致组织结合的自身抗体与BMZ分离。补体片段在补体激活期间不仅在IgG上被激活,而且在接近IgG的细胞的细胞表面上被激活。在炎症引起的低pH条件下,IgG可能会从BMZ中脱离。但BMZ还保留了一些补体片段.这些现象可能有助于解释为什么当DIF用于诊断BP时C3比IgG更敏感。
    Bullous pemphigoid (BP), an autoimmune subepidermal blistering disease, shows tense blisters associated with urticarial erythema. Tissue-bound Immunoglobulin G (IgG) at the basement membrane zone (BMZ) detected by direct immunofluorescence (DIF) is strong evidence for a diagnosis of BP. The sensitivity of DIF is higher in complement component 3 (C3) than in IgG, but the reason for this different sensitivity is not fully understood. In this study, we performed several ex vivo studies to investigate the possible mechanism of IgG negativity and C3 positivity at the BMZ by DIF in some BP cases. First, sera from BP patients showing IgG negativity by DIF were found to clearly react to the BMZ in their own DIF skin samples. Next, indirect immunofluorescence (IIF) was performed using sera diluted with different pH phosphate-buffered saline (PBS), pH 7.4, 6.0, and 3.0. Patients\' sera diluted with pH 7.4 PBS showed linear staining at the BMZ, but sera diluted with pH 6.0 PBS and pH 3.0 PBS showed lower fluorescence intensities. Finally, sections of skin from BP patients were pre-incubated with different pH PBS (pH 3.0, 6.0, and 7.4), followed by staining with anti-human IgG and C3. The fluorescence intensities were notably lower for IgG and C3 that had been pre-incubated with pH 3.0 PBS and pH 6.0 PBS than for IgG and C3 that had been pre-incubated with pH 7.4 PBS. These results suggest that a low pH condition hinders the binding of autoantibodies to the BMZ, that is, the drop in tissue pH induced by inflammation inhibits autoantibodies from depositing at the BMZ. Furthermore, the drop in tissue pH causes tissue-bound autoantibodies to detach from the BMZ. Complement fragments are activated not only on IgG but also on the cell surface of cells close to IgG during complement activation. IgG may detach from the BMZ under low pH condition induced by inflammation, but some complement fragments remain at the BMZ. These phenomena may help to explain why C3 is more sensitive than IgG when DIF is used to diagnose BP.
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  • 文章类型: Journal Article
    背景:天疱疮是一组皮肤大疱性病症,其特征在于在表皮的细胞间连接中存在的自身抗体的形成。诊断是通过临床,组织病理学检查,DIF。由于DIF需要冷冻切片,荧光标记抗体,检查用紫外线显微镜,和训练有素的人员,它的不可用性使明确的诊断具有挑战性。
    目的:评估补体和Ig的IHC染色在天疱疮病例中的实用性。
    方法:对26例确诊的天疱疮患者采用抗IgG单克隆抗体进行过氧化物酶免疫组织化学染色。IgA,IgM,IgG4、C3、C4d,以DAB为色原。天疱疮病例包括20例寻常型天疱疮(PV),4例天疱疮(PF),和两个天疱疮素食者(Pveg)。阳性被定义为Ig的沉积和互补的不同,细胞间连接处角质形成细胞的连续棕色染色。
    结果:在IHC上,共20个PV17显示IgG阳性(85%),11(55%)C4d,19(95%)C3d,和16(80%)IgG4沉积在表皮的细胞间连接。所有PF病例均显示IgG沉积物,三例(75%)IgG4,C3d,C4d两例Pveg均显示IgG和C4d阳性,而一例IgG4和C3d阴性。总体IgG,天疱疮的C3、IgG4和C4d表达占88%,88%,76.9%,61.5%的病例。这些标记之间的关系,IgG和C3组合的相关性最好(P值=0.80)。对IgG的敏感性,IgG4和C3为77.8%,73%,和73%。
    结论:我们得出结论,IHC是诊断PV的有用工具,其IgG和C3d的敏感性最高。在我们几乎所有的病例中,IgG和C3d的组合可以取代DIF,因此,FFPE切片上的IHC可用作DIF的替代方法。
    BACKGROUND: Pemphigus is a group of bullous disorders of the skin characterized by the formation of autoantibodies present in the intercellular junction of the epidermis. Diagnosis is made by clinical, histopathological examination, and DIF. As DIF needs frozen sections, fluorescent tagged antibodies, UV light microscope for examination, and trained personnel, its non-availability makes a definitive diagnosis challenging.
    OBJECTIVE: To evaluate the utility of IHC staining of complements and Ig in cases of Pemphigus.
    METHODS: Twenty-six diagnosed cases of Pemphigus were stained by Peroxidase immunohistochemical method using monoclonal antibody to IgG, IgA, IgM, IgG4, C3, C4 d with DAB as chromogen. Pemphigus cases include twenty of pemphigus vulgaris (PV), four cases of pemphigus foliaceous (PF), and two of pemphigus vegetans (Pveg). Positivity was defined as the deposition of Ig and complements as distinct, continuous brown staining of keratinocytes at intercellular junctions.
    RESULTS: On IHC total of 20 PV 17 showed positivity (85%) for IgG, 11 (55%) C4d, 19 (95%) C3d, and 16 (80%) IgG4 deposits at the intercellular junction of the epidermis. All cases of PF showed a deposit of IgG, with three (75%) cases for IgG4, C3d, and C4d. Both cases of Pveg showed positivity for IgG and C4d while one case was negative for IgG4 and C3d. The overall IgG, C3, IgG4, and C4d expression for pemphigus was seen in 88%, 88%, 76.9%, and 61.5% of cases. The relation between these markers, combination of IgG and C3, was best related to each other ( P value = 0.80). The sensitivities for IgG, IgG4, and C3 were 77.8%%, 73%, and 73% resp.
    CONCLUSIONS: We conclude that IHC is a useful tool in the diagnosis of PV with the highest sensitivity of IgG and C3d. The combination of IgG and C3d could replace the DIF in almost all of our cases, so IHC on FFPE sections be used as an alternative method to DIF.
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  • 文章类型: Observational Study
    最近,膜增殖性肾小球肾炎已根据替代补体途径和免疫荧光沉积的致病作用,分为免疫复合物介导的(ICMMPGN)疾病(由经典补体途径驱动)和补体介导的(C3GN)疾病(由替代补体途径驱动)。拟议的重组为治疗MPGN疾病提供了治疗和预后支持。
    本研究基于组织病理学和DIF检查来研究MPGN的模式,并将病例分为主要补体显性和免疫复合物介导的疾病,以获得更好的预后和治疗效用。
    这是一项在三级护理中心进行的为期2年的前瞻性观察性研究。临床疑似病例MPGN进行组织病理学和直接免疫荧光检查(DIF),根据补体介导的和免疫复合物介导的MPGN解释了这一发现。
    在620个肾活检中,36例患者的组织病理学和DIF均证实了MPGN的诊断,占所有活检的5.8%。根据DIF的发现,各组包括20例(55.6%)免疫复合物沉积,C3优势图片的11(30.5%),和5(13.9%)无免疫沉积物。关于DIF模式的分析,C3+Ig组16例(80%)和C3GN组6例(54.5%)以MPGN模式为主。新月体肾小球肾炎,全球肾小球硬化,C3GN组明显可见间质纤维化。
    DIF在处理MPGN病例中具有巨大的预后和治疗价值。
    UNASSIGNED: Membranoproliferative glomerulonephritis has in the recent past been regrouped into immune complex-mediated (ICM MPGN) disease (driven by the classical complement pathway) and complement-mediated (C3GN) disease (driven by the alternative complement pathway) based on pathogenetic role of alternative complement pathway and immunofluorescence deposits. The proposed regrouping lent therapeutic and prognostic support in managing the disease of MPGN.
    UNASSIGNED: The present study is undertaken to study the patterns of MPGN based on histopathological and DIF examination and sub-categorize the cases into mainly complement dominant and immune complex-mediated diseases for better prognostic and therapeutic utility.
    UNASSIGNED: This is a prospective observational study carried out in a tertiary care center over a period of 2 yrs. The clinically suspected cases of MPGN were subjected to histopathologic and direct immunofluorescence examination (DIF), and the findings were interpreted in light of complement-mediated and immune complex-mediated MPGN.
    UNASSIGNED: Out of 620 renal biopsies, diagnosis of MPGN was confirmed both on histopathology and DIF in 36 cases accounting for 5.8% of all biopsies. Based on DIF findings, the various groups comprised 20 cases (55.6%) of immune complex deposits, 11 (30.5%) of C3 dominant picture, and 5 (13.9%) of Nil immune deposits. On analysis of the patterns on DIF, 16 cases (80%) of C3 + Ig group and 6 (54.5%) of C3GN group showed predominantly MPGN pattern. Crescentic glomerulonephritis, global glomerulosclerosis, and interstitial fibrosis were markedly observed in C3GN group.
    UNASSIGNED: DIF is of immense prognostic and therapeutic value in managing cases of MPGN.
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