Dermatomyositis Which Was Double Positive for Anti-MDA5 and Anti-ARS Antibodies That Was Successfully Treated by Intensive Immunosuppressive Therapy

Myositis-specific autoantibody is associated with the clinical phenotype and prognosis of dermatomyositis. Anti-melanoma differentiation-associated gene 5 (MDA5) and anti-aminoacyl-tRNA synthetase (ARS) antibodies are generally mutually exclusive. We herein present an extremely rare case of dermatomyositis which showed double positivity for anti-MDA5 and anti-ARS antibodies. There have been very few reported cases of double positive anti-MDA5, anti-ARS antibodies. In such cases, the clinical characteristics of each autoantibody can coexist. Thus, we should pay attention to the rapidly progressing features of anti-MDA5 as well as the chronic relapsing features of anti-ARS for the better management of this rare condition.     

Anti-MDA5 antibody dermatomyositis-associated rapidly progressive interstitial lung disease patient complicated with mixed connective tissue disease: A case report

Anti-MDA5 antibody dermatomyositis (DM) is a special type of myositis, which can potentially cause rapidly progressive interstitial lung disease (RP-ILD). Mixed connective tissue disease (MCTD) is a complex disease with different characteristics of autoimmune connective tissue disease, associated with ILD. Both are rare diseases, and few patients with both diseases have been reported. A 71-year-old woman complained of palpitations, with a 2 months history of rash around her hands, extensor surface of right elbow, and the nape of her neck. Subsequently, the patient had acute exacerbation of dyspnea and tachypnea. Anti-Ro52, U1 RNP and MDA5 antibodies were positive; the presenting evidence was suggestive of anti-MDA5⁺DM-RP-ILD complicated with MCTD. Our patient deteriorated rapidly and had a fatal outcome, despite “triple therapy” for RP-ILD. This case illustrates that patients with coexisting anti-MDA5⁺DM and MCTD have the former's typical clinical manifestations, and may develop ILD quickly rather than slowly as in MCTD, especially with the coexistence of anti-Ro52 antibodies.     

Soluble CD206 levels correlate with disease deterioration and predict prognosis of anti-MDA5 antibody-positive dermatomyositis related interstitial lung disease

Introduction

The prognosis of anti-MDA5 antibody-positive dermatomyositis/clinically amyopathic dermatomyositis-associated interstitial lung disease (MDA5-DM/CADM-ILD) is poor. This study was to evaluate the effect of serum soluble CD206 (sCD206), a biomarker of macrophage activation, on predicting the interstitial lung disease (ILD) deterioration and prognosis for MDA5-DM/CADM-ILD.

Methods

Forty-one patients diagnosed with MDA5-DM/CADM-ILD were retrospectively included. The clinical data were analyzed. Serum sCD206 levels were measured in 41 patients and 30 healthy controls. The relation between sCD206 levels and ILD deterioration was assessed. Receiver operating characteristic (ROC) curve was generated to determine the optimal cut-off value of sCD206 for predicting outcome. The association between sCD206 and survival was examined.

Results

The median serum sCD206 level in patients was significantly higher than healthy controls (464.1 ng/mL vs. 349.1 ng/mL, P = 0.002). In DM/CADM patients, the sCD206 level was significantly higher in patients with acute/subacute interstitial lung disease (AILD/SILD) than those with chronic interstitial lung disease (CILD) (539.2 ng/mL vs. 309.4 ng/mL, P = 0.005). The AUC of sCD206 was 0.885 for predicting mortality (95% CI 0.779–0.990). Patients were divided into two groups: sCD206 high level group (≥400 ng/mL) and sCD206 low level group (<400 ng/mL). Patients with sCD206 high level had significantly decreased survival rate than those with low level (25% vs. 88%, P < 0.001). The adjusted hazard ratio of sCD206 for mortality was 1.003 (adjusted for age and gender, P < 0.001), with sCD206 high level associated with higher death risk (HR 4.857, P = 0.006).

Conclusion

Serum sCD206 might be a potential predictor of ILD deterioration and prognosis for Chinese patients with MDA5-DM/CADM-ILD.     

A competitive enzyme-linked immunosorbent assay for diminazene

Diminazene aceturate has remained a very important therapeutic drug for trypanosomosis in cattle, sheep and goats since its introduction into the market in 1955. Despite its continued use, the methods available for its detection in body fluids are lengthy and inefficient for routine monitoring of drug levels in treated animals. A competitive enzyme linked immunosorbent assay (ELISA) has now been developed and optimized for the detection of diminazene in bovine serum. In the assay, diminazene in the test samples and that in a newly developed diminazene-horseradish peroxidase conjugate compete for antibodies to diminazene raised in rabbits and immobilized on a microtitre plate. Tetramethylbenzidine-hydrogen peroxide (TMB/H(2)O(2)) is used as chromogen-substrate system. The assay has a detection limit of 0.8 ng/ml of serum with a high specificity for diminazene. Cross-reactivity with either homidium bromide and quinapyramine sulphate/chloride of 0.0004% is negligible while that with isometamidium chloride is 0.71%. The assay was able to detect diminazene levels in normal Boran steers for at least two weeks after intramuscular injection with the drug at a dose of 3.5 mg/kg bw. The assay will be useful in monitoring diminazene use, and development of resistance in trypanosomosis endemic areas.     

结核γ-干扰素酶联免疫斑点检测在结核性心包积液诊断中的应用

目的 探讨结核菌特异性γ-干扰素（interferon-gamma,IFN-γ）酶联免疫斑点（enzyme-linked immunosorbent spot,ELISPOT）检测对结核性心包积液的诊断价值.方法 采用结核菌特异性IFN-γ ELISPOT技术同时检测20例结核性心包积液患者（TP组）和14例非结核性心包积液患者（Non-TP组）外周血单核细胞（peripheral blood mononuclear cells,PBMC）及心包积液单核细胞（pleural effusion mononuclear cells,PEMC）中结核菌特异性IFN-γ水平.结果 TP组PBMC和PEMC结核菌特异性IFN-γ水平均显著高于Non-TP组,差异有统计学意义（P＜0.01,P＜0.01）.TP组心包积液中结核菌特异性IFN-y水平显著高于外周血IFN-γ水平,差异有统计学意义（P＜0.01,P＜0.01）.PBMC ELISPOT检测结核性心包积液的敏感性和特异性分别为80.0％和85.7％;而PEMC ELISPOT敏感性和特异性为90.0％和85.7％.结论 结核菌特异性IFN-γELISPOT技术对结核性心包积液诊断和鉴别诊断具有很好的辅助价值.     

Subacute cutaneous lupus erythematosus,and positive anti-MDA5 antibody in clinically amyopathic dermatomyositis with rapidly progressive interstitial lung disease: A case report and literature review

BackgroundClinically amyopathic dermatomyositis (CADM) is a subtype of DM with characteristic cutaneous lesion with normal creatinine kinase levels. Presence of anti-melanoma differentiation-associated gene 5 (MDA5) antibody is found to be associated with rapidly progressive interstitial lung disease (RP-ILD). Aim of the work: to report a CADM patient with positive anti-MDA5 antibody and RP-ILD with clinical features of systemic lupus erythematosus (SLE) who clinically responded to a combination of cyclophosphamide and other conventional immunosuppressant. Case presentation: A 44-year-old Indian woman presented with classical cutaneous lesions of DM with normal creatinine kinase levels amongst other clinical features. She was initially diagnosed with SLE before developing RP-ILD and a positive MDA5 antibody. Anti-nuclear antibody, anti-dsDNA and anti-Sm antibody were negative. Serum ferritin level was very high (1599 ng/mL) as compared to C-reactive protein (23.4 mg/L). Anti-Ro-52 and anti-PM-Scl 75 were positive. High resolution computed tomography (HRCT) of the lungs revealed features consistent with ILD. Histology of her skin biopsy was consistent with subacute cutaneous lupus erythematosus (SCLE). Her diagnosis was revised to CADM with overlapping SCLE. She responded to a combination of hydroxychloroquine, cyclosporine-A, mycophenolate mofetil, pulse methylprednisolone and pulse cyclophosphamide 750 mg/month for 6 months. Her cutaneous lesions gradually improved with normalization of serum ferritin level. Repeated HRCT showed no further progression of the pulmonary fibrosis. Conclusion: CADM with positive anti-MDA5 antibody associated with RP-ILD is rare with a high mortality rate. Early recognition and prompt treatment with a combination of immuno-suppressant may improve the outcome of this complex disease.     

Cardiac involvement in a case of juvenile dermatomyositis with positive anti-melanoma differentiation associated protein 5 antibody

Juvenile dermatomyositis (JDM) is an autoimmune disease characterized by muscle weakness and specific skin lesions, as well as non-muscular involvement such as interstitial lung disease (ILD), cardiac involvement and arthritis. Anti-melanoma differentiation-associated protein 5 (anti-MDA5)-positive JDM patients are typically characterized by the presence of skin ulcers and rapidly progressing ILD (RP-ILD). Although cardiac involvement is not an expected involvement in anti-MDA5-positive JDM cases, it is significant because it can be fatal. In this report, an anti-MDA5 myositis-specific autoantibody-positive JDM case referred with the diagnosis of psoriatic arthritis in whom cardiomyopathy and arrhythmia were detected in follow-up is presented. Since cardiac involvement is associated with mortality, it would be useful to follow up anti-MDA5 positive patients for cardiac involvement in addition to lung involvement. Tofacitinib is a promising treatment option in treatment-resistant JDM.     

Combined use of an antigen and antibody detection enzyme-linked immunosorbent assay for cysticercosis as tools in an epidemiological study of epilepsy in Burundi

OBJECTIVE: To evaluate the benefits of the detection of both circulating antibodies (Ab) and antigens (Ag) for the diagnosis of cysticercosis in people with epilepsy. Neurocysticercosis is a cause of neurological diseases world-wide, especially epilepsy. The clinical symptoms of neurocysticercosis are non-specific and diagnosis is often difficult. METHODS: Serum samples were collected from subjects in a matched case-control study for epilepsy in the Kiremba area, Burundi, between March and April 2001 (epileptic cases=303; controls without epilepsy=606). The enzyme-linked immunosorbent assay (ELISA) was used for the detection of antibodies (Ab-ELISA) and circulating Ag (Ag-ELISA). RESULTS: The Ab-ELISA revealed 58.7% positivity in epilepsy cases and 31.4% in healthy controls; and Ag-ELISA revealed 38.3% positivity in epilepsy cases and 20.0% in controls. The matched odds ratios were 3.6 (95% CI: 2.5-4.9) for Ab-ELISA, and 2.9 (95% CI: 2.1-4.3) for Ag-ELISA. CONCLUSION: Both Ag- and Ab-ELISA detected a significantly higher number of seropositives among people with epilepsy than among controls. The risk of epilepsy was high in cases with a positive Ag-ELISA, although less important than in cases with positivity for Ab-ELISA. Dead or degenerating cysticerci appear to be more frequently associated with epilepsy than living cysts. The high number of people with circulating Ag of Taenia solium suggests that the study area is a focus of active transmission of the parasite.     

检测T-2毒素的直接竞争性酶联免疫吸附测定法的研究

目的本研究在自制T-2毒素酶标半抗原的基础上,结合市售抗T-2毒素单克隆抗体,建立了检测谷物中T-2毒素的直接竞争性酶联免疫吸附测定法(ELISA)。方法粮食样品经70%甲醇-水溶液提取,用滤纸过滤并稀释后,再用0.45μm滤器过滤净化,该滤液用于ELISA检测。结果本方法最低检出量为0.125ng/ml,添加70~280ng/g T-2毒素的大米样品和面粉样品的平均回收率分别为85~117.5%和98.1~102.5%。结论结果表明,该方法简化了粮样提取步骤,可用于谷物及加工产品中T-2毒素的检测。     

双单克隆抗体夹心酶联免疫吸附试验检测鼠疫F1抗原的实验观察

目的 观察双单克隆抗体(F1-McAb)夹心酶联免疫试验(DMcAbS-ELISA)快速检测鼠疫F1抗原的敏感性和特异性.方法 采用鼠疫细菌学检验、DMcAbS-ELISA和反向间接血球凝集试验(RIHA)对比检测鼠疫感染鼠和阴性对照鼠脏器标本.结果 共检测225份阴性对照鼠脏器标本,鼠疫细菌学检验、DMcAbS-ELISA和RIHA法检测F1抗原均为阴性.共检测308只鼠疫感染鼠脏器标本,鼠疫细菌学检验、DMcAbS-ELISA、RIHA法阳性率分别为92.21％(284/308)、90.91％(280/308)和89.61％(276/308),3种方法比较,差异无统计学意义(x2=5.65,P＞0.05).DMcAbS-ELISA法与鼠疫细菌学检验结果符合率为97.00％[(274+243)/533],Kappa值为0.940;与RIHA法符合率为99.25％[(276+253)/533],Kappa值为0.985.脏器标本F1抗原检测的真实性比较:DMcAbS-ELISA法敏感性为96.48％(274/284),特异性为97.59％(243/249),阳性预测值为97.86％(274/280),阴性预测值为96.05％(243/253),一致性为96.99％11/4×(274/280+274/284+ 243/253+243/249)|,Youden指数为0.9407;RIHA法的敏感性为96.13％(273/284),特异性为98.80％(246/249),阳性预测值为98.91％(273/276),阴性预测值为95.72％(246/257),一致性为97.39％[1/4×(273/276+273/284+246/257+246/249)],Youden指数为0.9492.DMcAbS-ELISA法对鼠疫菌检测灵敏度为2.7×104cfu/ml,RIHA法为2.2×105 cfu/ml;两种方法检测F1抗原灵敏度均为10 μg/L.结论 DMcAbS-ELISA法检测鼠疫F1抗原具有敏感、特异、简便、快速的特点,是有应用价值的鼠疫快速诊断技术.     

Clinical application of an enzyme-linked immunosorbent assay detecting hepatoma-specific γ-glutamyltransferase

Aim:  To develop a sandwich enzyme-linked immunosorbent assay (ELISA) measuring hepatoma-specific datura stramonium agglutinin–tightly bounding γ -glutamyltransferase (DSA-GGT) and evaluate its clinical application for hepatocellular carcinoma (HCC) diagnosis.
Methods:  Serum DSA-GGT concentrations were measured with the sandwich ELISA system in 96 patients with HCC, 240 patients with chronic liver diseases and 119 healthy subjects. The diagnostic performance of DSA-GGT for HCC was assessed using receiver operating characteristic (ROC) curves. The diagnostic accuracy of DSA-GGT was compared with serum α-fetoprotein (AFP).
Results:  The area under the ROC curve of DSA-GGT in discriminating patients with HCC from non-HCC was 0.865 (95% confidence interval: 0.818–0.915, P  < 0.001). Serum DSA-GGT was positive in 67 out of 96 patients with HCC and 23 out of 240 patients with non-HCC diseases. The sensitivity and specificity of DSA-GGT and AFP for the diagnosis of HCC were 69.8% and 90.5%, and 72.9% and 89.1%, respectively. A higher sensitivity (93.8%) in the identification of HCC was observed by combining DSA-GGT and AFP.
Conclusion:  The sandwich ELISA system showed good reliability and reproducibility, and using the measurement, we found that serum DSA-GGT was a valuable marker of HCC, as a usable complementary to AFP. The sensitivity for identifying HCC could be significantly improved by combining DSA-GGT and AFP, and the combination could be used in large-scale screening for HCC in susceptible individuals.     

Spontaneous pneumomediastinum in a dermatomyositis patient with anti‐melanoma differentiation‐associated gene‐5 antibody and interstitial lung disease despite an initial response to immunosuppressant

We report a 24‐year‐old man with anti‐melanoma differentiation‐associated gene‐5 (MDA5) antibody‐positive dermatomyositis (DM) and interstitial lung disease (ILD) who developed spontaneous pneumomediastinum. By comparing serial thoracic high‐resolution computed tomography scans, we demonstrated the distinct time course showing a paradoxical occurrence of pneumomediastinum despite a radiological improvement of ILD. Our case shows that pneumomediastinum in DM can occur regardless of associated ILD and it is a serious complication that should be considered in DM patients presenting with pulmonary manifestations. Cutaneous vasculopathy may be associated with pneumomediastinum and could potentially be a useful indicator of future disease.     

贝氏柯克斯体5种重组表面蛋白抗原包被酶联免疫吸附试验的特异性和敏感性分析

目的 使用贝氏柯克斯体5种重组表面蛋白作为包被抗原的酶联免疫吸附试验用于血清学检测的特异性和敏感性分析。方法 本研究建立由贝氏柯克斯体的Com1、GroEL、Mip、OmpH、RplL重组表面蛋白抗原分别包被的酶联免疫吸附试验(ELISA)方法,然后分别检测贝氏柯克斯体实验感染小鼠血清和Q热患者血清中IgG特异性抗体。结果 8份贝氏柯克斯体感染小鼠血清中除1份血清与RplL反应为阴性外其它均为阳性;11份Q热患者血清中除1份血清与RplL、1份血清与OmpH反应为阴性外,其它血清反应均为阳性。将这5种重组蛋白分别与莫氏立克次体、黑龙江立克次体、恙虫病东方体实验感染小鼠血清反应,除1份黑龙江立克次体感染小鼠血清与GroEL反应为阳性外,其它血清反应均为阴性。另外,Com1对斑疹伤寒、斑点热、恙虫病患者血清的阳性反应率平均为22.2%,GroEL为25.0%,Mip为25.0%,OmpH为25.0%,RplL为13.9%。结论 这些结果证明这5种重组蛋白抗原均可被贝氏柯克斯体感染血清所识别。所建立的ELISA方法具有良好特异性和敏感性,可用作Q热血清学诊断。     

梅毒螺旋体优势表位抗原嵌合表达构建双抗原夹心法的研究

目的用基因工程方法表达嵌合的梅毒螺旋体优势表位抗原,建立检测血清梅毒抗体的双抗原夹心酶联免疫方法(double antigen sandwich enzyme-linked immunosorbent assay,DAS-EIA)。方法通过计算机软件分析选择梅毒螺旋体优势抗原表位,用聚合酶链式反应(PCR)扩增优势表位基因,构建了梅毒螺旋体多优势表位嵌合抗原(rTpN15-TpN17-TpN47)表达载体,转化宿主菌BL21(DE3)进行表达,亲和层析柱法纯化获得高纯度融合抗原,并用其建立检测梅毒抗体的DAS-EIA。结果表达的优势表位嵌合抗原具有很好的抗原性。用其建立的嵌合抗原DAS-EIA检测确诊的50份阳性和30份阴性血,阳性检出率和阴性检出率都是100%。结论嵌合抗原DAS-EIA法具有比间接EIA和重组单抗原DAS-EIA更高的灵敏度和检出正确率,其检测水平已经达到国外TPHA的水平。该方法的建立为临床检测梅毒开辟了新的方法。     

Painless Panniculitis upon the Treatment of Clinically Amyopathic Dermatomyositis with Anti-MDA5 Antibody

Panniculitis, a rare cutaneous manifestation in patients with dermatomyositis (DM), usually presents as a painful erythematous lesion. We herein report a 32-year-old woman with panniculitis that appeared as an indurated plaque without pain or redness after a 4-month episode of clinically amyopathic DM during treatment with prednisolone and tacrolimus. She experienced no pain; however, the firmness and extent gradually worsened. Based on our findings, including the histopathological results, DM panniculitis was diagnosed. Azathioprine was additionally administered, leading to remission. DM panniculitis can develop as a painless induration during immunosuppressive treatment, and azathioprine may be a useful treatment.     

结核杆菌特异性酶联免疫斑点技术在HIV感染人群中的应用

我国艾滋病病毒(HIV)感染人群中结核病的发病率高,造成了严重的疾病负担。提高潜伏结核和活动性结核的诊断准确性,对控制HIV病人中结核病的治疗和控制至关重要。该文总结了近年来出现、发展并完善的一种新的结核诊断技术,即结核杆菌特异性酶联免疫斑点技术,及其在HIV人群中的相关研究结果,并着重对该技术在潜伏结核和活动性结核的诊断、治疗效果监测以及结核相关性免疫重建炎症反应综合征中的应用进行了评价。     

粗提抗原检测炭疽血清抗体ELISA方法的初步评价

目的 对使用粗提抗原检测炭疽血清抗体的酶联免疫吸附试验(ELISA)方法进行初步评价.方法 用间接ELISA方法检测人群血清(健康人血清42份、炭疽病人血清42份)特异性抗体,用阳性血清对照绘制标准曲线,按照标准曲线计算出每份血清标本的抗体相对含量,所得结果与重组致死因子(rLF)方法的检测结果进行比较.结果 病人组血清抗体相对含量中位数为1.19,健康人组血清抗体相对含量中位数为0.24,两组比较差异有统计学意义(uc=7.643,P＜0.05).粗提抗原检测与rLF检测结果并不完全对应,但两种方法显示出较高的一致性.结论 粗提抗原检测炭疽血清抗体的方法能区分大部分的病人和健康人,有一定的应用潜力,可用在炭疽疾病监测工作中.