杀菌试验在伤寒早期诊断中的应用价值

本文用微量杀菌试验测定伤寒患者血清中杀菌抗体,作为伤寒早期诊断依据。检测早期伤寒患者21例,阳性20例。19例非伤寒发热病人,20例既往伤寒患者,153例正常人群对照,杀菌试验均为阴性。本试验可比肥达氏反应早3—10天出现阳性反应,并将伤寒早期诊断阳性率33.33%提高到95.24%。     

在96孔板中进行抗脂质过氧化的微量测定

以Fe²⁺/半胱氨酸诱导大鼠肝微粒体为基本模型,根据硫代巴比妥酸(TBA)反应原理,优化不同反应条件,建立了一种在96孔板上进行抗脂质过氧化测定的一步反应方法,该方法的灵敏度不低于传统的试管法,而且还具有微量、快速、简便等优点,特别适用于大规模筛选和研究抗氧化剂.此外,也可用于其它系统诱导的抗脂质过氧化的测定.     

微量板细胞病变法滴定水痘疫苗的研究

建立了用微量板细胞病变法滴定水痘疫苗的实验方法。将适宜稀释度的病毒与2BS细胞混种于96孔板培养,种毒后第7天观察孔内细胞病变情况,以karber法计算疫苗滴度,并与蚀斑法进行比较。实验表明,微量板细胞病变法滴定水痘疫苗,结果稳定性好,精密度高,较现行的蚀斑法操作简便、误差小,能够较好的反映出水痘的真实滴度,可作为一种较为简便的滴定水痘疫苗的方法。     

伤寒杆菌感染后IgG和IgM抗体消长规律的初步观察

本文报道了以试管凝集、SpA菌体吸收和2ME处理血清的方法对感染动物和部分伤寒病人血清中的IgG和IgM抗体的消长规律的观察结果。初次注射伤寒杆菌在3～30天中均为IgM抗体,再次注射后仍然以IgM抗体首先回升,然后才有IgG抗体的出现。在再次注射后所观察的期间,IgM抗体仍然维持较高的滴度。在观察的10例伤寒病人(6至30日病程)中,其抗体类型也是IgM。伤寒病人的血清学诊断,目前主要用肥达氏反应。本文以试管凝集、SpA菌体吸收和2ME处理血清的方法对感染动物血清和部分伤寒病人血清中的IgG和IgM抗体的消长规律进行了初步观察。     

微柱凝胶技术诊断伤寒及副伤寒的临床应用

目的 比较微柱凝胶技术(MGT)与肥达反应法(WR)在诊断伤寒及副伤寒中的意义.方法 采用微柱凝胶技术及肥达反应法分别检测317例疑似伤寒及副伤寒患者.结果 317例疑似患者中,通过对已被临床确诊或得到血培养和(或)粪便培养阳性报告的患者血清标本检测,微柱凝胶法要比肥达反应法平均提高1～2个滴度.统计学差异有显著性.结论 微柱凝胶技术检测伤寒杆菌抗体方法简便快速,灵敏度高,便于观察,能为临床早期诊断,快速诊断提供重要的参考依据.     

H5亚型禽流感病毒单抗-生物素捕获ELISA的建立

目的建立一种单克隆抗体介导的、经生物素—亲和素系统放大的H5亚型禽流感病毒捕获ELISA检测方法,为进一步研究检测试剂盒提供基础。方法用亲和层析法纯化抗禽流感病毒H5亚型血凝素单克隆抗体,包被微量反应板,用于捕获病毒抗原,再用生物素标记的单抗和酶标亲和素来检测病毒血凝素抗原,经方阵试验优化ELISA反应体系。用该方法检测H1—H15亚型AIV标准毒株和H5、H7、H9亚型AIV分离株,并与血凝和血凝抑制试验比较,评价其敏感性和特异性。结果纯化后的单抗具有良好的反应活性,生物素标记单抗工作浓度为1∶5000;ELISA对H5亚型AIV的检出限为025个血凝单位。该ELISA反应体系能检出H5N3标准株和所有20株国内H5亚型AIV分离株,而与其他14个血凝素亚型的AIV标准株、15个H9亚型AIV分离株和2个H7亚型AIV分离株均无交叉反应。结论初步建立了检测H5亚型禽流感病毒的单抗—生物素捕获ELISA方法,为研制试剂盒和进一步应用试验提供了基础。     

一种测定蛹虫草中虫草酸含量的酶标仪微量反应方法

研究和建立一种基于酶标仪-96孔板高通量测定虫草酸含量的检测方法,并对该方法进行性能评价。以酶标仪为检测仪器,在96孔板内按照设定反应条件微量加入样品和试剂进行显色反应,利用酶标仪测定吸光度值并计算虫草酸含量。通过检测精密度、重复性、回收率,并与分光光度计法进行比较,综合评价该方法的准确度、精确度。结果表明,测定数据具有较高的精密度（样品CM1的RSD值0.829%;样品CM2的RSD值1.772%)、重复性（标准样品B40的RSD值2.061%;样品CM2 的RSD值1.599%)、回收率（平均回收率99.24%,RSD值3.666%）,测定结果与分光光度法检测结果无显著差异(P>0.05)。结果表明,酶标仪微量法测定准确、重复性好,并可大大减少样品和试剂的用量,该方法方便、快捷、高效,可以替代分光光度法用于虫草酸含量的测定。     

032 布鲁菌病和伤寒血清学试验试剂条的评价

伤寒和布鲁菌病常引起血液感染。血及骨髓培养是最准确的诊断方法,在许多发展中国家,这两种疾病的诊断常凭临床表现和经验。血清学试验常被作为一种诊断工具,但肥达反应和布鲁菌试管凝集……     

感染伤寒期间抗体对沙门氏伤寒菌脂多糖和蛋白质抗原的反应——Ⅰ、放射免疫测定血清抗体

作者收集24名成人血培养伤寒菌阳性病人血清及20名健康成人对照血清,用固相放射免疫测定对伤寒菌脂多糖和蛋白质抗原的反应,并用传统的肥达氏试验进行了比较。抗原为新分离伤寒菌培养于心脑浸液琼脂上一夜,用盐水洗下培养物,加     

简易快速酶标免疫吸附试验

酶标免疫吸附试验(FLISA)具有敏感、准确等特点。国内通用的微量板法(Microplate)因受到仪器及器材限制,影响一步推广。为此,作者建立一个简易快速酶标免疫吸附试验方法(SR—ELISA)。本法不用分光光度计,不用微量板,不用终止反应。作者用此法测定了肝吸虫病人抗体,为了观察其敏感性及准确性,在测定时间使用了微量板 ELISA及琼脂扩散 ELISA(DIG—ELISA)进行了对比。结果说明 SR—ELISA 与微量板 ELISA 及 DIG—ELISA 具有相同的敏感性,但在操作上更为简易,可在4小     

Evaluation of rapid methods for differentiation of Bifidobacterium species

R oy , D. & W ard , P. 1990. Evaluation of rapid methods for differentiation of Bifidobacterium species. Journal of Applied Bacteriology 69 , 739–749.
The fermentation patterns of 20 Bifidobacterium strains were determined by two methods: a carbohydrate utilization test, based on the production of acid from a given range of sugars in modified MRS broth (micromethod), and a gas chromato-graphic (GC) analysis of fermented modified MRS broth. The micromethod provides a qualitative fermentation pattern whereas the GC method gives a quantitative evaluation of carbohydrate utilization. The micromethod and GC analysis can be applied for the differentiation of Bifidobacterium strains used in fermented milks.     

A microassay for tetrazolium-reductase activity of polymorphonuclear leukocytes--comparison with a test-tube technique

A micromethod for quantitative INT test is described to assay the activity of human peripheral blood PMNL. PMNL are obtained from a pellet after the separation of mononuclear cells. The test is performed in microtiter plates with 96 wells. The estimation of the activity is based on the colour change following the INT reduction to formazane. As a stimulating agent starch (rice species--Amylum oryzae) or zymosan is used. Absorbance alternations are measured using a microplate reader (e.g. MR 580, Dynatech). Conditions necessary for standard results (i.e. PMNL concentration, type and concentration of stimulating agent, incubation time, sedimentation time for PMNL isolation) were verified experimentally. Blood samples of 60 blood donors were simultaneously tested by the micromethod and the test-tube method. The statistical analysis (F-test of homogeneity of variances, t-test with paired observations, correlation coefficient of tightness of dependence) proved that the results of the micromethod were comparable to the test-tube assay. The advantages of the micromethod are as follows: low blood consumption, possibility to perform more parallel examinations of the same sample, and to test more samples at a time, standard and highly reproducible results, saving of reagents, reduction of labour time, possibility of some further automation.     

Quantitative analysis of cholesterol in 5 to 20 microliter of plasma

A gas-liquid chromatographic micromethod for quantitation of cholesterol in 20 micro l of plasma was developed using 5alpha-cholestane as an internal standard, saponification with tetramethylammonium hydroxide-isopropanol, and extraction with tetrachloroethylene-methyl butyrate. Cholesterol levels in plasma samples were calculated by comparing cholesterol-cholestane peak height ratios with those of preassayed reference plasma. Over a plasma cholesterol range of 44 to 468 mg/100 ml, the gas-liquid chromatographic micromethod and the automated ferric chloride colorimetric method gave nearly identical results (r = 0.99) in duplicate aliquots of 131 plasma samples.     

Microtiter Plate Agglutination Test for Brucella canis Antibodies

A micro-agglutination test for the antibodies to Brucella canis produced similar results to those obtained with the standard tube agglutination method in human and canine sera. The micromethod does provide an economical means of screening sera for the presence of antibodies.     

Micromethod for Serogrouping Escherichia coli

A micromethod for serogrouping strains of Escherichia coli is described. The method is accurate, rapid, and economical in terms of time and reagents utilized to perform the test.     

A micromethod for agglutination and antiglobulin tests for antibody to Brucella abortus

A micromethod has been developed for the standard agglutination and antihuman globulin tests for serum antibodies to Brucella abortus. The test uses a haematoxylin-stained antigen. The test is simple and quick to perform and, in a comparison at two centres involving 367 sera, the micromethod was found to be more sensitive than the classical test. It is highly suitable both for routine serological screening and for large-scale surveys of occupationally exposed industrial groups.     

Micromethod of determining antibodies to nerve tissue galactocerebrosides in the complement fixation reaction

A micromethod has been developed for the determination of serum antibodies to galactocerebroside, a specific antigen of myelin membranes. The method is based on complement fixation test and is used for the analysis of sera from healthy rabbits and those immunized with galactocerebroside.     

A method for determinations of microgram amounts of chitin in arthropod cuticles

A method is described for determining microgram amounts of chitin in arthropod cuticles. The method, adapted from one described for assaying plant material for fungal content, is based on estimating soluble chitosan derivatives and is not affected by the presence of nonchitinous cuticular components. There is good agreement between the results given by the micromethod and those given by a semimicro method based on material insoluble in hot, dilute alkali. As a qualitative test for chitin the limit for the method is <100 ng.     

Usefulness of the passive hemagglutination micromethod in the diagnosis of whooping cough

Studies on modification of diagnostic test for pertussis have been continued, they were practically restricted to an application of passive haemagglutination micromethod. Six hundred and twenty eight sera from children suspected to be infected with Bordetella and 38 sera of control children suffering from non-infectious diseases were tested. Despite of the fact that higher titers were obtained with method using red blood cells preserved in Alsevier solution, the passive haemagglutination micromethod may be used in field studies especially during pertussis epidemics what was confirmed by statistical analysis.