Detection of tubercular antibody and antigen in sera of bone and joint tuberculosis

Trichloroacetic acid (TCA) solubilized and DEAE fractionatedMycobacterium tuberculosis H₃₇Ra excretory-secretory (ES) antigen viz., Mtb EST DE1 and affinity purified goat antibodies to the TCA solubilized ES antigen (Mtb EST) were explored in detecting tubercular antibody and antigen respectively in sera of bone and joint tuberculosis by indirect and sandwich ELISA. Out of total 36 bone & joint tuberculosis cases, tubercular antibody was detected by indirect ELISA in 30 patients (sensitivity 83%), while circulating tubercular antigen was detected by sandwich ELISA in 27 patients (sensitivity 75%). Out of 34 non tubercular disease control cases, 10 patients showed positive reaction for antibody while only 4 patients showed positive reaction for antigen. In another group of 34 healthy subjects who were screened, 4 individuals showed positive reaction for tubercular antibody and 2 cases for antigen. This study shows that antigen detection assay using affinity purified anti Mtb EST antigen antibody is superior with overall specificity of 91% as compared to antibody detection assay with 75% specificity in bone & joint tuberculosis.     

新烟碱杀虫剂噻虫嗪免疫学特性研究

通过化学修饰合成了噻虫嗪（thiamethoxam,TMX）人工半抗原,采用碳二亚胺法（EDC）将该抗原与牛血清蛋白和卵清蛋白偶联成功制备了分子结合比合理的免疫抗原（TMX-BSA）和包被抗原（TMX-OVA）.对经TMX-BSA免疫的6周龄Balb/c实验鼠的免疫抗血清进行了间接ELISA和阻断ELISA,初步明确了抗噻虫嗪多克隆抗体（TMX-pAb）的免疫学特性,为噻虫嗪残留免疫学快速检测技术开发和检测产品研制奠定了坚实基础.     

青少年花样游泳运动员身体素质“敏感期”内的体能训练

在分析花样游泳项目体能需求和青少年体能素质发展"敏感期"的一般规律的基础上,阐述了在体能带训中的实践方法,并提出了目前实践中所面临的主要问题。旨在进一步探讨符合项目培养规律的青少年体能训练体系,并为将来的深入研究提供一些参考和依据。     

Passengers information in public transport and privacy: Can anonymous tickets prevent tracking?

Modern public transportation companies often record large amounts of information. Privacy can be safeguarded by discarding nominal tickets, or introducing anonymization techniques. But is anonymity at all possible when everything is recorded? In this paper we discuss travel information management in the public transport scenario and we present a revealing case study (relative to the city of Cesena, Italy), showing that even anonymous 10-ride bus tickets may betray a user's privacy expectations. We also propose a number of recommendations for the design and management of public transport information systems, aimed at preserving the users’ privacy, while retaining the useful analysis features enabled by the e-ticketing technology.     

Establishment of Trimester-Specific Reference Intervals of Serum TSH & fT4 in a Pregnant Indian Population at North Kolkata

Reference intervals (RIs) of serum thyroid stimulating hormone (TSH) and free thyroxine (fT₄) were determined in 402 healthy pregnant women by enzyme-linked immunosorbent assay (ELISA) technique after partitioning them into three trimesters. The reference population was chosen from a study population of 610 pregnant females by applying strict inclusion and exclusion criteria. The assays were done using proper quality control measures. RIs were calculated from the central 95 % of the distribution of TSH and fT₄ values located between the lower reference limit of 2.5 percentile and upper reference limit of 97.5 percentile value 0.90 confidence intervals for the upper and lower reference limits were also determined. The reference intervals for TSH were 0.25–3.35 μIU/ml for the first trimester; 0.78–4.96 μIU/ml for the second trimester and 0.89–4.6 μIU/ml for the third trimester. Similarly, the reference intervals for fT₄ for first, second and third trimesters were 0.64–2.0, 0.53–2.12 and 0.64–1.98 ng/dl respectively. The values thus obtained varied from those provided by the kit literature. In comparison to our derived reference intervals, the reference data from kit manufacturer under-diagnosed both subclinical hypo- and hyper-thyroidism within our pregnant reference population.     

植物病毒抗血清的制备及IgG纯化与酶标技术研究

以烟草花叶病毒（TMv）为例研究了制备植物病毒抗血清的一般方法．以家兔和小白鼠为免疫动物，运用饱和硫酸接沉淀法制备出高效价的兔和瓦抗TMV血清，并运用先进的ELISA方法测定了抗血清中抗体IgG的效价；用醋酸纤维凝胶电泳技术分析了抗血清中抗体的lgG纯度，应用免疫双扩散法检测了提纯的IgG的生物活性．同时还运用成二醛一步法制出高结合率的IgG与过氧化氢酶（HRP）标记的抗体，形成了一整套制备抗血清、纯化IgG并对IgG进行酶标的技术，为植物病毒检测提供了快速简便的方法．     

ICA69基因工程融合抗原的制备

基于基因工程技术 ,用PCR法扩增出编码ICA6 9的cDNA片段 ,直接克隆到pSPORT 1质粒上 ,经DNA序列测定 ,插入到GST融合蛋白表达载体 pGEX 2T ,构成重组质粒 p2T ICA6 9,得到的表达产物GST ICA6 9融合蛋白用间接ELISA法检测其免疫原性 .测序结果表明 ,所获PCR产物已正确重组到PGEX 2T表达型质粒中 .重组质粒在原核细胞中表达的融合蛋白具有免疫原性 ,并能应用于Ⅰ型糖尿病病人血清中抗ICA6 9抗体的检测 .所获得的表达产物为重组ICA6 9融合抗原 ,有助于提高Ⅰ型糖尿病的预报率和确诊率 .     

Isolation ofMycobacterium Tuberculosis 31 kDa antigen protein of diagnostic interest from culture filtrate using anti-ES-31 antibody by affinity chromatography

Proteins secreted into the culture medium byMycobacterium tuberculosis (M. tb) are shown to be source of antigens of immunodiagnostic interest. Anin vitro released 31 kDa antigen ESAS-7F isolated fromM.tb H₃₇Ra culture filtrate by salt precipitation, SDS-PAGE and cation exchange fast protein liquid chromatography (FPLC) was shown earlier to be a diagnostically important antigen fraction. In this report, we describe the isolation of ESAS-7F antigen using monospecific antibody coupled to sepharose CL-4B column. The percentage recovery of ESAS-7F antigen using affinity chromatography was approximately 8% of the total ES antigen proteins compared to 0.05% obtained by conventional purification steps using salt precipitation, SDS-PAGE and FPLC. Similar seroreactivity was observed by the antigen isolated by both the methods in indirect ELISA. Affinity chromatography helped in an increased recovery of ESAS-7F antigen and obviates the need for time consuming conventional purification steps.     

ELISA技术在饲料安全检测中的应用

近年来随着兽药在饲料中的使用日益广泛,饲料安全检测工作在畜牧生产中更加重要,具有高选择性、高灵敏度、高分析效率、低成本、低环境污染特点的ELISA技术成为当今饲料安全检测工作中应用日益广泛的测定技术之一．ELISA技术在饲料安全检测中的运用及其试验条件的要求就显得非常重要．     

Diagnostic role of the antibody response to the 38kDa, 16kDa proteins and lipoarabinomannan of mycobacterium tuberculosis

The antibody response to the 38kDa, 16kDa and Lipoarabinomannan (LAM) antigens ofMycobacterium tuberculosis was evaluated using three different ELISAs based on these antigens. The study group included tuberculosis patients (n=52), patients with HIV and TB co-infection (n=10), other chest symptomatics (n=5), HIV infected individuals (n=10), leprosy cases (n=7) and healthy controls (n=75). The results indicate that the 38kDa and LAM based ELISA for IgM/IgG has a low specificity (ranging from 69–85%) and sensitivity (ranging from 55–78%). When three ELISAs are carried out on a single patient the probability of detection of tuberculosis was significantly increased to 95.2% indicating that a single ELISA test is of low sensitivity and that a combination of ELISA’s may be needed to be of any value as a diagnostic test for tuberculosis. Additionally, a western blot assay of the serum antibody response to protein fraction ofM. tuberculosis was analysed in 15 tuberculosis patients and five healthy controls. A multiple antibody response to various M.tuberculosis proteins was observed which varied from patient to patient as compared to controls who showed a single 38–39 kDa protein band positivity. These finding suggest that a western blot assay which determines the antibody response to a set of antigenic components ofM. tuberculosis could be a better serological test for the diagnosis of tuberculosis in our population.