一种抗破伤风类毒素的保护性人单克隆抗体

人类淋巴细胞分离的单克隆抗体在治疗和诊断上具有各种潜在用途。破伤风类毒素曾被用作模式抗原,因而一种具有这种特异性的人类抗体曾在临床应用。在企图制备杂种细胞过程中,作者取来自一个免疫供体的淋巴细胞和一株对HAT敏感的类淋巴母细胞系间进行,一种自发的转化引起分泌破伤风类毒素特异性抗体,该细胞系命名为ES12,具有44条染色体,     

产生人抗破伤风毒素杂交瘤抗体

用破伤风毒素免疫过的人B淋巴细胞与鼠骨髓瘤细胞进行融合,产生的种间杂文瘤,能分泌人抗破伤风毒素抗体。本实验的一株杂交克隆,在培养十三个月后,仍继续分泌高滴度的抗破伤风毒素。该抗体能结合破伤风毒素的B段,而且能保护小鼠免受破伤风毒素的侵害。此实验研究,将提供用人的单先隆杂交瘤抗体来预防和治疗传染病的方法。     

深圳市1003名外来劳务工破伤风抗体水平的调查

目的利用横断面基线调查法对深圳市1003名工厂劳务工破伤风抗体水平进行调查研究,制定相应的免疫接种措施,加强对外来劳务工的管理。方法采用酶联免疫吸附试验检测破伤风抗体。结果深圳市1003名工厂外来劳务工来自全国28个省市自治区,抗体阳性率较高的人群主要集中在中部、南部和西南部。1003名外来劳务工破伤风抗体的阳性率为18.6%,其中男性为18.7%,女性为18.4%。29岁以下劳务工破伤风抗体的阳性率为15.7%,30～39岁的阳性率为2.1%,40岁以上的阳性率为0.8%。小学文化及以下劳务工破伤风抗体阳性率为14.8%,初中文化为14.7%,高中或中专文化为21.0%,专科为21.6%,大学本科及以上文化为26.5%。结论深圳市工厂劳务工破伤风抗体水平偏低。抗体阳性率在不同年龄组间存在显著差异,29岁以下劳务工破伤风抗体的阳性率较高。建议政府为外来劳务工进行破伤风疫苗的普种,以进一步提高和保持高水平的百白破混合制剂常规免疫接种率,同时加强外来流动人口免疫接种的监测,防止破伤风病的发生。     

酶免疫法(EIA)检测乙型肝炎e抗体

<正> 乙型肝炎e抗原、e抗体的检测在临床疾病预后、流行病学分析及疫苗安全性试验等方面均具有重要的意义。1981年我们建立了酶免疫法检测乙型肝炎e抗原。随后,在酶标记抗HBe IgG方面我们建立了一种简便、快速、高效的过碘酸钠法,提高了酶的标记率,在这些工作基础上建立了下述检测乙型肝炎e抗体(抗HBe)的方法。     

两种配制破伤风抗毒素皮试方法的比较观察

为预防破伤风的发生;对外伤、烧伤、动物咬伤常规注射破伤风抗毒素.破伤风抗毒素是经胃酶消化的马破伤风免疫球蛋白,具有抗原性,注射后易出现过敏反应.因此在使用前需做破伤风皮内试验.通过两种不同的配制方法可以看出传统配制法与该法配制是有区别的.破伤风的皮试液的配制在临床中应用广泛.该法值得推广.     

McAb ELISA间接夹心法检测HFRS病人血清IgM和IgG抗体的研究

肾综合征出血热(HFRS)早期临床症状不典型,因此在病人血清中检出特异性抗体,特别是IgM抗体对辅助临床诊断有重要意义。此外,HFRS的流行病学监测和病原学研究等都需有敏感、特异、简便而客观的     

白喉和破伤风类毒素抗原ELISA检测方法的建立

目的建立定量检测白喉和破伤风类毒素抗原的酶联免疫吸附试验(ELISA)方法。方法分别采用白喉、破伤风单抗包被酶标板,兔抗白喉、破伤风多抗作为二抗,辣根过氧化物酶标记的羊抗兔IgG抗体作为酶标抗体,以平行线法建立定量检测白喉和破伤风类毒素抗原含量的夹心ELISA法,并进行方法学验证。结果两种定量检测ELISA方法的验证结果均符合规定。检测白喉类毒素抗原ELISA方法的定量限为8.90×10-4Lf/ml,回收率为98.35%,批内变异系数(CV)≤10.59%,批间CV≤13.51%;检测破伤风类毒素抗原ELISA方法的定量限为2.13×10~(-3)Lf/ml,回收率为107.28%,批内CV≤13.96%,批间CV≤10.06%。结论建立了白喉、破伤风类毒素抗原ELISA检测方法,该法特异性强、准确度高、重复性好,可用于白喉破伤风疫苗产品的质量控制和生产过程控制。     

母亲、脐带及新生儿百日咳、白喉、破伤风抗体的效价测定

本文用ELISA法探讨母子之间百日咳、白喉,破伤风抗体效价移行情况,以及新生儿后期抗体效价下降的程度。结果表明,正常母体与脐带血中抗LPF、FHA、白喉、破伤风类毒素效价之间密切相关,而生后1个月的婴儿血中抗体效价明显低于脐血抗体。     

妊娠妇女麻疹、风疹和破伤风抗体及HBsAg携带率调查

为了解妊娠妇女麻疹、风疹和破伤风的免疫水平及乙肝表面抗原 (ＨＢｓＡｇ)的携带情况 ,随机检测了 131名妊娠早期体检妇女的血清 ,采用间接ＥＬＩＳＡ检测麻疹ＩｇＧ和风疹ＩｇＧ抗体 ,采用间接血凝试验检测破伤风凝集抗体 ,采用酶标方法检测ＨＢｓＡｇ。结果显示 ,麻疹和风疹抗体阳性率较高 ,分别为 97.7%和 95 .4 % ;麻疹抗体几何平均滴度 (ＧＭＴ)为 114 0 .4 ,水平较高 ,但仍有 19.1%为麻疹阴性或低于保护水平 ,高抗体 (≥ 1∶32 0 0 )的比例占38.9%。不同年龄组 (2 2～ 32岁 )抗体水平不同 ,有出疹史的抗体水平明显高于没有出疹史的 (…     

应用组合抗体文库技术制备人源单克隆抗体

<正>组合抗体文库(Combinatorial antibody frag-ment library)是基因工程抗体技术之一,使真正意义上的临床疾病的免疫预防与免疫治疗成为可能.本文就组合抗体文库的技术路线及应用前景作一概述.近年来,单克隆抗体的抗病毒研究获重要进展.抗体不仅可以中和病毒,阻止其在细胞间的传播,而且可以清除MHC-I缺陷小鼠体内已经建立的病毒感染(Diane,Criffin,Walter Gerhard),这意味着特异抗体在抗感染免疫中可能具有重要作用.1992年以来,国外学者首次应用分子生物学方法,将人的抗体编码基因克隆到噬菌体中并以工程菌表达出人的抗体活性片段(Fab),其抗原中和能力竟比人的自然抗体高出一千倍以上,具有极大的开发前景和社会意义.     

Comparison of ELISA and toxin neutralization for the determination of tetanus antibodies

In a sandwich ELISA for tetanus antibodies, the influence of the tetanus toxoid concentration used for coating microtiter plates has been studied. The antibody levels by toxin neutralization bioassay and by ELISA were studied for a population with known immunization history. By decreasing the tetanus toxoid concentration in ELISA from 5 to 0.2 Lf/ml, a better correlation was found between the ELISA results and the bioassay titers, but sera from recently immunized people still showed high ELISA titers. This phenomenon cannot be ascribed to nonspecific reactions since sera from nonimmunized people are negative in both assays. All sera negative in ELISA had, however, a bioassay titer beneath 0.01 IU/ml.     

Comparison of ELISA and Toxin Neutralization for the Determination of Tetanus Antibodies

In a sandwich ELISA for tetanus antibodies, the influence of the tetanus toxoid concentration used for coating microtiter plates has been studied. The antibody levels by toxin neutralization bio-assay and by ELISA were studied for a population with known immunization history. By decreasing the tetanus toxoid concentration in ELISA from 5 to 0.2 Lf/ml, a better correlation was found between the ELISA results and the bioassay titers, but sera from recently immunized people still showed high ELISA titers. This phenomenon cannot be ascribed to nonspecific reactions since sera from non-immunized people are negative in both assays. All sera negative in ELISA had, however, a bioassay titer beneath 0.01 IU/ml.     

单克隆抗体ELISA间接夹心法检测家鼠型HFRS疫区人和动物血清中特异性抗体

用McAb-ELISA间接夹心法和IFAT或酶标SPA染色法平行检测山西地区(家鼠型HFRS疫区)的人及动物血清中HFRS病毒特异性IgM和/或IgG抗体。结果ELISA检测174份HFRS病人血清的阳性率及抗体滴度均明显高于IFAT。检测295份无明确HFRS病史的健康人血清,ELISA的阳性率也高于IFAT。检测215份鼠类血清,102份兔血清及108份猪血清,ELISA的阳性检出率与IFAT或酶标SPA染色法基本相同。用阻断试验等证明本ELISA检出的确是HFRS病毒特异性抗体。对ELISA的某些试验条件作了讨论,并认为,McAb-ELISA在帮助临床早期诊断及血清流行病学调查等方面均有实用价值。     

Penicillin Hypersensitivity

An enzyme-linked immunosorbent assay for the detection of anti-pencillin antibodies of the several Ig classes is described. The results of the ELISA in 350 sera of patients suspected of penicillin hypersensitivity are compared with those of the haemagglutination test. In 105 sera penicillin-specific IgM and/or IgG was demonstrated with the ELISA, the HA test being positive in 49 of these 105 sera. However , in another 14 sera IgM anti-penicillin antibodies could be shown only with the HA. In 10 sera penicillin-specific IgE was demonstrated with the ELISA, only four of these being also positive with the RAST. IgE was always found in combination with IgG and/or IgM. The positive correlation of the ELISA and the RAST with the intracutaneous test on penicillolypolylysine was 26.9% and 15.4% respectively. The ELISA is a simple and reproducible method for the detection of anti-penicillin antibodies, being more sensitive than the HA and the RAST.     

放射免疫沉淀法与酶联免疫吸附试验检测抗-LSP的双盲对照研究

研究表明,抗肝细胞膜特异性脂蛋白抗体(抗-LSP)在病毒性肝炎,特别是慢性活动性肝炎的发病机理中可能起较重要的作用。关于抗-LSP的检测,国内外文献中多     

High incidence of type II autoantibodies in pernicious anaemia.

AIMS: To investigate the incidence of type II autoantibodies to intrinsic factor in pernicious anaemia. METHODS: Three hundred and forty four serum samples submitted for intrinsic factor antibody (IFAB) analysis on clinical or laboratory grounds were tested by an established radioassay and a new enzyme linked immunosorbent assay (ELISA) method for type I and total IFAB, respectively. Sixty of these were found to be positive by ELISA; this method was used to test further, 40 samples of adequate volume for types I and II antibodies. RESULTS: Type II antibodies were detected in 39 of the 40 sera tested. A comparative analysis indicated that seven samples contained pure type II antibody, being positive for total and type II by ELISA, but negative for type I by both the ELISA and radioassay technique. CONCLUSIONS: The occurrence of type II antibody, both alone and in combination with type I, seems to be more common than has previously been recognised, and emphasises the advantage of using a technique which will detect both types of antibody.     

检测抗HIV-1/2型抗体的双抗原夹心方法的建立及其试剂盒的研制

目的 建立更敏感的检测人免疫缺陷病毒（HIV）抗体的方法,并研制检测试剂盒。方法 根据HIV－1／2型的基因序列及其所编码氨基酸结构,采用固相法合成了HIV－1型的gp41.1、gp41.2、gp120、p24和HIV－2型的gp36五条多肽,混合包被酶标板作为固相抗原。用辣根过氧化物酶标记以上多肽抗原作为标记物,建立检测血清中抗HIV－1／2抗体的双抗原夹心ELISA法。同时,应用该方法制备检测HIV抗体的试剂盒,并检测三批中国卫生中药品和生物制品检定所HIV诊断试剂国家参比品。结果 建立了检测HIV－1／2抗体的双抗原夹心法。用检定所参比品检测,该方法特异性、灵敏度均为100％,变异系数小于10％。与间接法相比较其灵敏度、特异性均高于间接法（P＜0．05）。检测210份其他病种患者血清均为阴性。与GBI公司的HIV抗体诊断试剂比较,检测40份卫生部药品和生物制品检定所提供的质控参比品（阳性20份,阴性20份）,GBI试剂阴、阳性符合率及总符合率分别为100%(20/20)、85％（17／20）及92．5％（37／40）,而应用该方法所研制的诊断试剂盒、阳性符合率及总符合率为100％。该试剂已通过国家卫生部质检。与雅培公司HIV诊断试剂比较检测90份献血员血清和88份HIV－1／2型感染者血清,符合率为100％。试剂盒于37℃放置4d后的检测结果的阴、阳性判定不受影响。结论 本法特异性强、灵敏度高、稳定性好,适用于献血员的筛选和临床HIV感染的检测。     

Preparation and evaluation of recombinant severe fever with thrombocytopenia syndrome virus nucleocapsid protein for detection of total antibodies in human and animal sera by double-antigen sandwich enzyme-linked immunosorbent assay

The recent emergence of the human infection confirmed to be caused by severe fever with thrombocytopenia syndrome virus (SFTSV) in China is of global concern. Safe diagnostic immunoreagents for determination of human and animal seroprevalence in epidemiological investigations are urgently needed. This paper describes the cloning and expression of the nucleocapsid (N) protein of SFTSV. An N-protein-based double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) system was set up to detect the total antibodies in human and animal sera. We reasoned that as the double-antigen sandwich ELISA detected total antibodies with a higher sensitivity than traditional indirect ELISA, it could be used to detect SFTSV-specific antibodies from different animal species. The serum neutralization test was used to validate the performance of this ELISA system. All human and animal sera that tested positive in the neutralization test were also positive in the sandwich ELISA, and there was a high correlation between serum neutralizing titers and ELISA readings. Cross-reactivity was evaluated, and the system was found to be highly specific to SFTSV; all hantavirus- and dengue virus-confirmed patient samples were negative. SFTSV-confirmed human and animal sera from both Anhui and Hubei Provinces in China reacted with N protein in this ELISA, suggesting no major antigenic variation between geographically disparate virus isolates and the suitability of this assay in nationwide application. ELISA results showed that 3.6% of the human serum samples and 47.7% of the animal field serum samples were positive for SFTSV antibodies, indicating that SFTSV has circulated widely in China. This assay, which is simple to operate, poses no biohazard risk, does not require sophisticated equipment, and can be used in disease surveillance programs, particularly in the screening of large numbers of samples from various animal species.     

Specificity of antibodies directed against the cytolethal distending toxin of Haemophilus ducreyi in patients with chancroid

Antibodies specific for the cytolethal-distending toxin of Haemophilus ducreyi (HdCDT) complex and for the CdtA, CdtB, and CdtC components were measured by ELISA in the sera of 50 patients with culture and/or PCR proven chancroid, 42 patients with periodontitis, 50 blood donors from Tanzania, 50 blood donors from Sweden. In addition, the biological activity e.g. neutralization capacity of the sera were tested. Our results demonstrate that majority of chancroid patients and healthy individuals had detectable levels of serum antibodies to HdCDT complex and to separate toxin components. However, high levels (> or =100 units) of antibodies to HdCDT complex were significantly more prevalent in the sera of patients with both chancroid and periodontitis than in the sera of the corresponding controls (P=0.001 and P=0.04, respectively). In the sera of the 50 patients with chancroid, antibodies to CdtA, CdtB, and CdtC were detected in 50, 35, and 34 individuals, respectively. Antibodies to CdtC, being less frequently detected than the antibodies to other components, show a good correlation with the neutralizing capacity of sera. High levels of neutralizing antibodies (> or =160) were detected in only 22 and 2% of the patients with chancroid and periodontitis, respectively. The data suggest that the low levels of anti-HdCDT antibodies, which include neutralizing antibodies, may contribute to limited protection in chancroid and since anti-HdCDT antibodies, may be detected in healthy individuals and in patients with certain disease conditions (e.g. periodontitis), they may not be specific markers for chancroid infection.