免疫酶染色试验检测丝虫病患者血清IgG抗体

本文报告应用马来丝虫成虫冰冻切片抗原进行免疫酶染色试验,检测115例淋巴丝虫病微丝蚴血症患者血清IgC抗体,阳性率为93.9%;99例健康人血清假阳性率为2.02%;45例其它寄生虫病患者未出现交叉反应。同时观察到丝虫病患者抗体水平与微丝蚴密度无相关性。     

Trustline结核分枝杆菌IgG/IgM抗体检测试剂盒(胶体金法)的临床应用评价

目的 评价Trustline结核抗体IgG/IgM检测试剂盒临床应用效果。 方法 选用3家医院的1009份血清标本,其中628份为结核病患者血清(308例菌阳病例,320例菌阴病例);对照组381份非结核病血清。采用Trustline试剂盒及一种已上市的试剂盒(对照试剂盒)分别检测1009份血清,计算多项检测指标,从而评价试剂盒的检测效果。 结果 通过检测1009份临床血清标本, Trustline试剂盒检测血清抗体(IgG+IgM)的灵敏度、特异度、阳性预测值、阴性预测值、 Youden 指数分别为61.3%、79.8%、 83.3%、 55.6% 和0.411,对照试剂盒检测血清抗体(IgG) 的结果分别为53.7%、89.0%、88.9%、 53.8%和0.426。经统计分析表明Trustline试剂盒的灵敏度显著优于对照试剂盒(P0.01),特异度低于对照试剂盒(P0.01),但阳性诊断效率和阴性诊断效率方面差异无统计学意义,Youden指数相接近。 进一步分析显示Trustline试剂盒对菌阳样本及菌阴样本的检出率分别77.6%和44.7%,对照试剂盒则为67.9%和40.0%,对菌阳样本检出率的差异有统计学意义。检测1009份标本,Trustline试剂盒共得IgM阳性35例,其中结核组阳性30例(4.8%),非结核组阳性5例(1.3%)。 结论 Trustline试剂盒检测结核抗体的灵敏度显著优于对照结核抗体试剂盒,并可同时用于检测IgG/IgM抗体,可以应用于结核病的快速诊断。     

冬桑叶注射液对微丝蚴血检阳性病例36例疗效观察

为摸索中草药解决海群生治疗丝虫病禁忌症问题,我们试用50%冬桑叶注射液治疗丝虫血检微丝蚴阳性病例36例,取得一定疗效。病例选择从晚上8时半至12时,采取耳垂血3     

酶联免疫吸附试验检测丝虫病人血清中的免疫复合物

本文报告用抗C_3和抗丝虫免疫球蛋白G(FSI-G)作酶联免疫吸附试验(ELISA)检测丝虫特异抗原抗体复合物。采集斑氏丝虫流行区(印度)丝虫病人(有微丝蚴血症或有临床表现如鞘膜积液和象皮肿)、无     

上海市南汇区丝虫病基本消除后的监测分析

目的了解和评价上海市南汇区淋巴丝虫病自1981年基本消除后的监测效果,为WHO将对中国消除丝虫病的情况进行认证准备充分的资料。方法采用病原学、蚊媒、免疫学方法对慢性丝虫病患者、原微丝蚴阳性者、外来流动人口、儿童、本地居民等进行监测。结果在丝虫病基本消灭后监测阶段(1982～1996年),病原学检测阳性率为0.003%,免疫学检测各类监测对象阳性率为0.04%～0.24%,儿童免疫学监测抗体阳性率为0.90%,蚊媒监测均为阴性;消灭后巩固监测阶段(1997年后),病原学检测阳性率为0,免疫学检测各类监测对象阳性率为0.07%￣0.46%,儿童免疫学监测抗体阳性率为0,蚊媒监测均为阴性。结论南汇区自1981年宣布基本消灭丝虫病以来,仅发现1例微丝蚴阳性者,1987年以来人群和蚊媒监测均没有发现微丝蚴阳性者,说明丝虫病的传播已经被阻断,监测成果是巩固的,但仍需做好监测工作。     

两种试剂盒检测TORCH-IgM的比较性研究

用自制试剂盒和HOPE试剂盒检测1196份孕妇血清TORCH-IgM,共检出阳性血清67份,在67份阳性血清中,有4份血清用自制试剂盒检测为阳性血清(A值虽大于临界值,但比较低,为0.215～0.231),而用HOPE试剂盒检测为阴性。所有这67份阳性血清均经其它试验证实为阳性。两者阳性、阴性的符合率分别为94%、99%。结果表明自制试剂盒是初筛孕妇血清TORCH-IgM的一个具有敏感、特异、简便等特点的良好工具。     

SARS康复者及医护人员冠状病毒的初步调查

目的　采用荧光聚合酶链反应 (F PCR)和基因芯片技术检测严重急性呼吸综合征 (SARS)冠状病毒 ,并探讨其临床应用价值。方法　应用 SARS Co V F PCR诊断试剂盒及基因芯片技术检测了 6 0份确诊 SARS患者血清、发热门诊医护人员血清 2 0份和漱口液样本 2 0份以及 1份 SARS疑似患者血清的 c DNA。结果　中山大学达安基因股份有限公司及上海复星实业股份有限公司的两种 F PCR检测试剂盒及晶宇芯片反应 3种检测方法所测 80份血清和 2 0份痰液样本均为阴性 ;但 1例 SARS疑似患者血清的 c DNA可经荧光定量 PCR反应扩增出病毒特异 RNA片段。结论　 SARS康复者及密切接触医护人员血液和漱口水中均未检测到 SARS病毒特异 RNA片段。     

一种定量PCR检测乙肝标记物假阴性的应对方法

【目的】了解定量PCR检测乙肝HBsAg(+)、HBeAg(+)及抗HBcAg(+)患者血清样本阴性结果中,采用不同厂家试剂盒对检测结果的影响。【方法】将定量PCR检测阴性患者血清样本用另一公司试剂进行检测。【结果】15例乙肝患者血清样本中有4例定量PCR检测结果小于1.0E+03copies/ml,被判为阴性结果;经换用其他试剂盒并重新检测后定量结果分别为2.8×103、1.9×104、1.1×105、1.2×106copies/ml,均为阳性结果。【结论】不同厂家检测试剂盒可能会对定量PCR的检测结果产生影响。     

血清乙肝病毒标志物和循环免疫复合物间关系

目的 探讨乙型肝炎患者血清HBV抗原抗体标志物和HBsAg特异及非特异性循环免疫复合物间的关系。方法 205份血清，用PEG沉淀法检测CIC，ELISA检测HBVM、HBs-CIC、HBsAg/C3-TCIC、IgG/C3-TCIC，免疫比浊法检测IgG和C4。结果 ①乙肝病毒感染过程中半数以上有HBsAg特异性的循环免疫复合物形成，HBsAg阳性特别是同时伴有HBeAg阳性时，血清各类CIC阳性率及IgG水平均显著增高，C4水平显著下降。②HBsAg阴性、抗HBs阳性血清中，各类CIC仍有13％－21．7％的阳性率。结论 乙肝病毒血清标志物的存在及模式与HBsAg特异及非特异性循环免疫复合物的形成有关。     

全自动粒子化学发光法在梅毒抗体筛查中的应用与分析

目的评价威高Autolumis 3000微粒子化学发光分析仪在梅毒特异性抗体筛查试验中的特异度。方法对梅毒特异性抗体的检测结果进行回顾性分析。采用威高Autolumis 3000梅毒特异性抗体检测试剂,对2018年3月至2019年3月大连医科大学附属威海市立医院36854例患者的血清标本进行检测。对初筛结果阳性标本附加甲苯胺红不加热血清试验(TRUST),再采用梅毒螺旋体明胶凝集试验(TPPA)进行复检,并对TPPA试验复检结果为阴性的样本采用免疫印迹法(Western blot)进行确证。采用SPSS软件进行检测结果的数据统计分析。结果36854份血清标本中,微粒子化学发光免疫法共初筛检出阳性标本544份。TRUST试验复检阳性314份,阴性230份。经TPPA试验复检后,检出阳性标本526份,阴性标本18份;对18份TPPA阴性结果的血清标本,采用Western blot试验确证,检测结果为阳性2份,不确定样本5份,阴性11份。Autolumis 3000微粒子化学发光法检测梅毒特异性抗体的特异度为99.97%。结论微粒子化学发光免疫法检测梅毒特异性抗体的特异度高,对初筛阳性标本选用TPPA试验作为补充试验进行复检,可有效提高梅毒检出的准确性。     

Antigen-specific immune complexes in urine of patients with lymphatic filariasis

Lymphatic filarial subjects with disease manifestations exhibit significantly elevated levels of immune complexes (ICs) in their circulation. The objective of the study was to explore the possible excretion of filaria-specific soluble ICs in urine of subjects with lymphatic filariasis. Paired urine (overnight) and serum samples were analyzed for complement activating filarial antigen containing immune complexes by enzyme-linked immunosorbent assay (ELISA). Antigen-specific ICs were detected in urine samples of 34% of subjects with filarial disease manifestations while the frequency of occurrence was low in microfilaremic subjects. The titer of urine ICs is significantly high in subjects with chronic filariasis as compared to microfilaria (mf) carriers. The occurrence of filaria-specific ICs in urine and their passage through the filtering structures of the kidney is suggestive of the focal or diffuse damage in those subjects. Detection of ICs in urine may provide a noninvasive means of assessing the extent of renal damage in patients with lymphatic filariasis.     

The development and evaluation of a single step multiplex PCR method for simultaneous detection of Brugia malayi and Wuchereria bancrofti

A single step novel multiplex polymerase chain reaction (PCR) has been developed for simultaneous detection of human filarial parasites, Brugia malayi and Wuchereria bancrofti, from blood samples and mosquitoes. The primers used were novel and have been tested with the parasite DNA amplifying 188bp (BM) and 129bp (WB) DNA fragments, specific to B. malayi and W. bancrofti, respectively, in a single reaction. The specificity of the PCR product was confirmed by DNA sequencing and slot blot hybridization assay. The test was found highly sensitive for both B. malayi and W. bancrofti by detecting the parasitaemia up to the level of one microfilaria per reaction. The assay was further evaluated on 98 blood samples and 144 mosquito samples collected from filarial endemic areas. The PCR was found to be more efficient in comparison to microscopy by detecting 8% and 5% more filarial parasites in field-collected blood and mosquito samples, respectively. This novel PCR that offers scope for simultaneous detection of both the parasites may be used as a diagnostic tool for the detection of filariasis in population and can be adopted for rapid surveillance and monitoring of mosquitoes for use in the effective control of filariasis.     

The “Filarial Dance” Is Not Characteristic of Filariasis

The objective of this series was to show that the sonographic appearance described as the “filarial dance” is not characteristic of filariasis but occurs in nonendemic areas as a manifestation of epididymal obstruction. An experienced observer documented cases after initial observation of the filarial dance in routine clinical practice using high‐frequency linear array transducers. The filarial dance was described as excessive to‐and‐fro movement of echogenic particles within a prominent epididymis and graded 1 to 4 according to the extent and distribution of the abnormality. The country of birth, exposure to filarial infection or travel to a filarial‐endemic area, previous scrotal surgery including vasectomy, any previous or current scrotal inflammatory disease, and any congenital testicular abnormalities were recorded. Over a 10‐year period, sonographic appearances consistent with the filarial dance were observed in 18 patients (bilateral in 6). The mean patient age was 47.7 (range, 28–91) years. The abnormality was graded in the 24 affected testes as follows: grade 1, n = 3; grade 2, n = 8; grade 3, n = 8; and grade 4, n = 5. No patient had a history of filariasis or travel to an endemic area. Six of 18 patients (33.3%) had bilateral vasectomies; 5 (27.8%) had a history of epididymo‐orchitis in the ipsilateral testis; 3 (16.7%) had previous scrotal surgery; and 4 (22.2%) had no relevant urologic history. We have described a sonographic appearance identical to the filarial dance in men with no history of filarial infection. Most had previous scrotal surgery or infection, suggesting that the filarial dance may not always be due to movement of filarial worms. The unifying condition in patients with filariasis and our patients is lymphatic obstruction, likely the underlying cause of the appearance in both groups.     

Whole blood samples as an alternative to serum for detection of immunity to measles virus by ELISA

The aim of this study was to evaluate enzyme linked immunosorbent assay (ELISA) as a testing strategy for detection of antibodies against measles virus from microquantities of blood soaked onto filter paper. We studied 165 healthy children in the age group of 1 to 2 years, attending the outpatient department of pediatrics. Two sets of samples were collected from each child. One by venipuncture and the other on Whatman filter paper-3 discs of 20 mm size by finger or heel prick so that each strip is completely soaked with blood on both sides. These were tested for measles virus antibodies by ELISA using Melotest measles IgG commercial ELISA kit manufactured by Melotec S. A. (Barcelona, Spain). The resulting sensitivity, specificity, positive predictive value (PPV) and negative predictive value (NPV) of the filter paper (FP) ELISA compared to serum ELISA was 100, 90, 97.8, and 100%, respectively. The correlation coefficient r = 0.93% (p < 0.001) and the agreement between the two techniques was 98% as calculated by the Kappa statistical method. The present study has found filter paper testing by ELISA to be a promising qualitative technique for detection of immunity against measles.     

Antigen detection assay with parasite specific monoclonal antibodies for diagnosis of lymphatic filariasis

BackgroundLymphatic filariasis is a painful and profoundly disfiguring disease. Infection is usually acquired in childhood but its visible manifestations occur later in life, causing temporary or permanent disability. The importance of developing effective assays to diagnose, monitor and evaluate human lymphatic filariasis has been emphasized by the WHO.MethodsHigh-affinity monoclonal antibodies (mAbs) specific for recombinant filarial antigen WbSXP-1 were developed. An ELISA based capture assay using monoclonal and polyclonal antibodies for WbSXP-1 was used for detection of circulating filarial antigen.ResultsHigh-affinity monoclonal antibodies (mAbs) were developed that specifically binds both W. bancrofti and B. malayi mf antigens. Two mAbs (1F6H3 and 2E12E3) of subclass IgG2a and IgM showed high affinity, avidity and reactivity to recombinant and mf native antigen. Both the mAbs were used in combination as capture antibodies and polyclonal as detection antibody to develop the assay. The assay showed very high sensitivity towards W. bancrofti mf positive samples compared to endemic normal samples (P < 0.0001).ConclusionA capture assay using high-affinity monoclonal antibodies for WbSXP-1 was developed for the detection of filarial circulating antigen in clinical samples from bancroftian infection. Besides, this would also help in epidemiological studies in endemic areas of filarial infections.     

不同方法检测乙肝表面抗原的结果分析

目的分析酶联免疫吸附法(ELISA)和电化学发光免疫法((ECLIA)检测乙肝表面抗原(HBsAg)的结果及临床应用价值。方法收集经ELISA法检测过的120份临床患者的血清,其中包括:30份HBsAg阳性;30份HBsAg阴性但乙肝e抗体(eAb)和乙肝核心抗体(cAb)阳性;30份HBsAg阴性但cAb阳性;30份乙肝5项均阴性。对上述血清采用ECLIA进行HBsAg检测,并对检测结果进行比较分析。结果 30份经ELISA检测HBsAg阳性者经ECLIA检测均为阳性,其灵敏度达100.0%;30份经ELISA检测HBsAg阴性而eAb、cAb阳性者中,有4份血清经ECLIA检测HBsAg呈阳性,其阳性率达13.3%;30份经ELISA检测HBsAg阴性而cAb阳性者中,有2份血清经ECLIA检测HBsAg呈阳性,其阳性率达6.7%;30份血清经ELISA检测乙肝5项均阴性者中,有2份血清经ECLIA检测HBsAg呈阳性,其阳性率达6.7%。结论 ECLIA法较ELISA法敏感性更高;对临床避免医院内有创检查、手术、输血等情况下的乙肝感染和传播等具有重要应用价值。     

Dance of live adult filarial worms is a reliable sign of scrotal filarial infection.

OBJECTIVE: To determine the value of the filarial dance sign as a diagnostic sign of scrotal filarial infection and to recognize unsuspected scrotal filariasis by this sign. METHODS: Eight symptomatic patients in whom the filarial dance sign was shown on high-resolution ultrasonography were studied, investigated, and followed after treatment with diethylcarbamazine citrate. Two patients underwent fine-needle aspiration. RESULTS: Multiple foci (nests) of motile (live) filarial worms were observed in most patients. Fine-needle aspiration of the dilated lymphatic vessels in 2 patients confirmed the presence of microfilariae. Five of 8 patients had a favorable response to treatment with diethylcarbamazine citrate. CONCLUSIONS: High-resolution ultrasonography is a useful technique for diagnosing scrotal filariasis in symptomatic patients and is very useful in the follow-up period for assessing the response of worms to treatment.     

Differential diagnosis of human lymphatic filariasis using PCR-RFLP

Filariasis is still a public health problem in tropical countries. The most common causative agents of human filariasis are Wuchereria bancrofti and Brugia malayi. Traditional methods used to detect filarial parasites in human, animal and vector populations are tedious, time consuming, and confer little guarantee of sensitivity and species specificity. We have developed a rapid and specific method to detect filarial parasite DNAs in blood and mosquito samples using the polymerase chain reaction (PCR) technique. The primers used are MF/F and MF/R which amplify a 1.5 kb glutathione peroxidase gene of filarial worms. Using the restriction fragment length polymorphism (RFLP) technique, these PCR products will be further digested with restriction enzymes either Hpa I, Pst I, Alu I or Hinf I to differentiate the genus of filaria. This PCR-RFLP technique can be apply to use in diagnosis and to differentiate between species of filaria in humans the reservoir host and the mosquito vector in endemic areas Copyright 2000 Academic Press.     

Immunoreactive molecules of Brugia malayi and their diagnostic potential

Antigen derived from three major life-stages of human lymphatic filariid, Brugia malayi was fractionated by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and reactivity of filarial antibodies, present in sera of human bancroftian patients belonging to different categories of the disease, to immunoreactive molecules was evaluated by Western blotting and immune recognition. Antigen molecules of >180, ~180, ~116, ~66, 58, 33 and <20 kDa were strongly identified in blots especially by symptomatic and endemic normal sera. Although mf positive serum reacted with the majority of these molecules, the intensity of bands was poor. These functional molecules along with ~43 kDa were later isolated from a preparative SDS-polyacrylamide gel by elution. The diagnostic utility of these purified molecules was then assessed by enzyme linked immunosorbent assay (ELISA) for detecting the IgG antibodies in patients' sera of various categories. Tropical pulmonary eosinophilic subjects revealed highest reactivity with <20 and ~116 kDa molecules while mf positive ones reacted feebly with the majority of the molecules except ~43 and ~66 kDa. Sera of endemic normals revealed high IgG levels but antibodies to ~ 66 kDa were highest. Symptomatic patients showed moderate reactivity. Non-endemic sera neither reacted in blots nor in ELISA. The study demonstrates the usefulness of ~43 kDa in detecting IgG antibodies of mf positive asymptomatic patients. High IgG levels to ~66 kDa followed by ~116 kDa and <20 kDa in sera of endemic normals warrants their immunoprophylactic evaluation.     

肠道病毒71型手足口病ELISA诊断试剂盒研制与临床应用

目的 研制早期快速检测抗肠道病毒71型(EV71)抗体的ELISA血清学诊断性试剂盒,评价其临床应用价值.方法 将表达纯化的EV71重组蛋白VP1作为包被抗原,建立EV71型手足口病间接ELISA的血清学检测方法 .通过与逆转录(RT)PCR方法 、EV71病毒分离试验和微量血清中和试验比较,评价抗.EV71 IgM和抗-EV71 IgG血清学诊断方法 在EV71型手足口病的诊断价值.结果 与RT-PCR比较,抗-EV71 IgM敏感度、特异度、阳性预测值和阴性预测值分别为83%、85%、81%和87%;抗.EV71 IgG敏感度、特异度、阳性预测值和阴性预测值分别为72%、74%、68%和77%.与EV71病毒分离方法 比较,抗-EV71 IgM敏感度、特异度分别为85%和97%;抗-EV71 IgG敏感度、特异度分别为75%和77%.通过直线相关分析发现,抗-EV71 IgG抗体滴度和中和抗体滴度呈显著正相关(r=0.72,P＜0.05).EV71型手足口病患儿恢复期血清抗IgG滴度较急性期升高(P＜0.01),但抗IgM滴度差异无统计学意义(P＞0.05).结论 利用VP1重组蛋白作为包被抗原,成功开发ELISA检测人血清抗-EV71 IgM和抗-EV71 IgG抗体的诊断试剂盒.