犬细小病毒血凝的检测和血凝抑制试验方法的优化

目的优化血凝和血凝抑制（HA／HI）试验条件,提高HA／HI试验的稳定性和准确性。方法在不同的缓冲液、pH值、猪红细胞浓度、BSA浓度下进行HA／HI试验,选择合适的HA／HI条件。结果pH7．0、0．1％BSA、0．2mol／LPBS、1％猪红细胞可作为HA／HI试验合适的反应条件,猪血球可保存12d不影响试验结果。结论方法优化后稳定性增强、检测时间缩短、准确性提高。     

用反向被动血凝及血凝抑制方法检测流行性出血热病毒抗原和抗体

本文建立了检测流行性出血热病毒(EHFV)液体抗原及抗体的反向被动血凝(RPHA)和血凝抑制(RPHI)方法,RPHA检测EHFV抗原的敏感性与ELISA相近;RPHI检测EHFV抗体与IFA的符合率为97.3％,敏感性略低。     

猴源细小病毒血凝和血凝抑制试验的优化与应用

目的优化猴源细小病毒的血凝和血凝抑制（HA/HI）试验条件,以优化的方法对160份2010年～2011年间采集于中国北京地区圈养猴群的血清样品进行猴源细小病毒抗体调查。方法 在不同的缓冲液、缓冲液pH值、猪红细胞浓度、兔血清浓度、阿氏液比例和温度下进行猴源细小病毒的HA/HI试验,选择合适的HA/HI条件。通过优化的HA/HI方法对160份猴血清进行猴源细小病毒抗体检测。结果pH 7.0、0.02 mol/L PBS、0.75%猪红细胞、0.5%兔血清和4℃可作为猴源细小病毒HA/HI试验合适的反应条件,猪血球以1∶1阿氏液抗凝,可保存5～7 d不影响实验结果。猴血清的猴源细小病毒抗体阳性率为58.1%。结论方法优化后稳定性增强、检测时间缩短、准确性提高。细小病毒在我国北京地区圈养猴群中感染比较普遍。     

双歧杆菌血凝特性的研究

观察3株双歧杆菌对人O型、人B型、绵羊、兔、鸡和小鼠红细胞的凝集滴度,并对其血凝素进行了提取。结果表明,3株双歧杆菌均能凝集各种来源的红细胞,血凝滴度无明显差异;D-甘露糖不能抑制血凝。提取的脂磷壁酸(LTA)具有血凝活性。其血凝素受体为糖类。     

麻疹病毒血凝素基因H的点突变与血凝作用的转变

经Ｂ９５ａ细胞系纯化分离的三株麻疹病毒Ｆｕ、ＩＭＡ、ＳＭＤ的血凝试验（ＨＡＤ）为阴性,但将它们分别感染Ｖｅｒｏ细胞并传代培养后其血凝试验由阴性转变为阳性。实验证明这种血凝表型的转变与血凝素基因Ｈ的表达与吸附无关。以ＰＣＲ扩增血凝素基因Ｈ和融合基因Ｆ,并分别进行序列分析。表明三株病毒在两种培养细胞中传代后其融合基因Ｆ的序列未发现差异。但血凝素基因Ｈ的序列却有显著的点突变并伴随血凝表型的转变。其中Ｆｕ     

虎血清犬腺病毒抗体调查

犬腺病毒（Canine adenovirus,CAV）属于腺病毒科哺乳动物腺病毒属成员,分为犬腺病毒Ⅰ型（CAV-1）和犬腺病毒Ⅱ型（CAV-2）,其中CAV-1主要引起狐狸（Vulpes vulpes）脑炎和犬（Canis familiaris）传染性肝炎（殷震和刘景华,1997;胡体拉,1963）。在国内,夏咸柱等（1984）首次分离到CAV-1,随后,多个地区的不同动物中又有相继分离到该病毒的报道（钟志宏等,1990;范泉水和袁国庆,1992;夏咸柱等,1984）。CAV-2主要引起犬的喉气管炎和幼犬咳嗽,在我国的感染也比较普遍（范泉水和夏咸柱,1999）。     

血凝法的生物载体选择与处理

用ＨＢｓＡｇＲＰＨＡ做为模式,选用羊、火鸡、鹅、人Ｏ型血球为载体,醛化后分成致敏和不致敏两种,与异嗜性抗体、中国生物制品药品检定所的参比品等作凝集试验。结果表明,无论特异性还是灵敏度都以人Ｏ型血球为最佳。在此基础上,设计了五种醛化方法处理人Ｏ型血球,共得出１８个醛化样品,经７３种不同的比较试验,筛选出最佳醛化方法为改良丙酮醛──甲醛法,采用该法醛化后血球为褐色,阴性血清和空白对照背景清晰,血球沉集点致密,下滑程度好。致敏抗—ＨＢｓＭｃＡｂ后,使ＨＢｓＡｇ灵敏度达到１～２ｎｇ／ｍｌ。为制备高质量的血凝试剂提供了最佳载体。     

被动血凝试验测定伤寒Vi抗体

作者从拘橼酸杆菌中提取纯化获得其Ｖｉ多糖抗原,该抗原具有伤寒沙门氏菌Ｖｉ抗原的免疫学特性,而不含伤寒沙门氏菌０,Ｈ抗原,用其致敏新鲜羊血球作被动血凝试验,特异性敏感性均很好。所需Ｖｉ抗原致敏浓度极低,仅为０．０５ｕｇ／ｍｌ。使用新鲜羊血球凝模式较好,便于观察结果,采用被动血凝试验检测１０６名健康中学生肌肉注射３０ｕｇ伤寒Ｖｉ多糖菌苗前后Ｖｉ抗体的变化情况,发现免疫后血清抗体的四倍增长率达８９％,表明伤寒Ｖｉ多糖苗具有良好的免疫原性。     

Serological diagnosis of an infestation caused by the early developmental stages of botfly larvae (Oedemagena tarandi) in reindeer

Undertaken investigations have shown that the antigen prepared from larvae of O. tarandi is diagnostically effective and stricktly specific only to this infection in the reaction of indirect hemagglutination (RIHA). The experiments have proved the possibility of practical use of RIHA as a principal method of early diagnosis of infection of reindeer caused by O. tarandi.     

The standardization of the reverse indirect haemagglutination test for the assay of the viral antigen of Japanese encephalitis vaccine

The reverse indirect haemagglutination (RIHA) test has been standardized for the assay of the viral antigen of purified Japanese encephalitis (JE) vaccine. Glutaraldehyde fixed sheep erythrocytes were sensitized with ammonium sulphate purified antibodies to JE vaccine raised in mice. The sensitivity of the erythrocytes fell to about one hundredth of the initial sensitivity in the first two days after preparation. After initial loss in sensitivity the stability of the cells became stabilized and the cells retained their titre for one year at 4-8 degrees C. The initial loss in sensitivity was not reduced by storing the cells at -70 degrees C, but after freeze drying the sensitized cells with a stabilizer one day after their preparation the cells retained their sensitivity. The RIHA test has been found to be a highly reproducible and sensitive method for detecting viral antigen in 5-10 ng of protein nitrogen. The sensitivity of the test was affected by the origins of the erythrocytes, i.e. from the different sheep from which they were drawn. To obtain results more rapidly, goose erythrocytes were used in place of sheep erythrocytes and the sensitized goose erythrocytes gave RIHA results in only 40 min.     

Influenza A Neuraminidase Antibody Assay with Sensitized Erythrocytes

Erythrocytes sensitized with purified neuraminidase (Hong Kong) antigens were used for assay of influenza A neuraminidase antibodies. The neuraminidase indirect hemagglutination test was equal to the neuraminidase hemagglutination-inhibition (enhancement) test and appeared to be better than the neuraminidase inhibition test for detection of fourfold or greater antibody rises in paired sera from influenza patients or vaccinees. It was better than both tests for detection of neuraminidase antibody. The neuraminidase indirect hemagglutination test is simple to perform and has the advantage of direct antigen-antibody assay.     

Use of antisera against bovine (NCDV) and simian (SA11) rotaviruses in ELISA to detect different types of human rotavirus.

Two ELISA systems for the detection of human rotaviruses were developed. In the first system antibodies to Nebraska calf diarrhea virus (NCDV) were used for coating the solid matrix and for the preparation of the enzyme conjugate. In the second system antibodies to human rotavirus and antibodies to simian rotavirus (SA11) were used for coating the solid matrix and for the preparation of the enzyme conjugate respectively. The second ELISA system proved to have a broader spectrum for the detection of human rotaviruses. By using the two ELISA systems, the different types of human rotavirus could be distinguished. The ELISA tests developed were 8 to 64 times as sensitive as electron microscopy (EM) and (or) counterimmunoelectrophoresis (CIEP). The antigen detected by ELISA was shown to be different from that detected by the hemagglutination test.     

Comparison of the Direct Agglutination and Indirect Hemagglutination Tests in the Determination of Blood Serum Titers to Escherichia coli Organisms

A comparison of the direct agglutination test and the indirect hemagglutination test for the detection of blood serum antibodies to Escherichia coli organisms indicated that these serological tests were comparable. In some instances the indirect hemagglutination test provided higher endpoint readings. Preparation of the antigens for the indirect hemagglutination test was more time consuming than for the direct agglutination test. Crude extract and purified polysaccharides were comparable as red blood cell sensitizing agents.     

Comparison of the Reliability and Sensitivity of Three Serological Procedures in Detecting Antibody to Yersinia pestis (Pasteurella pestis)

Three serological procedures, the agar-gel precipitin inhibition, the complement fixation, and the indirect hemagglutination tests, were used to detect and measure antibody to Yersinia pestis in the sera from 383 individuals. Although all three tests were useful in detecting plague antibody, the most reliable and sensitive test procedure was indirect hemagglutination.     

Vero细胞法检测白喉抗毒素的研究

参照Ｍｉｙａｍｕｒａ等报道,建立了微量细胞培养检测白喉抗毒素的方法。以家兔皮肤试验为参照,Ｖｅｒｏ细胞培养法敏感度为９８．１１％,特异度为８４．００％,符合率为９６．２０％,相关系数ｒ＝０．９３,在白喉血清抗毒素测定中值得推广     

Use of the indirect hemagglutination inhibition reaction for evaluating the specific activity of allergens

To evaluate the specific activity of house-dust allergen, the passive hemagglutination inhibition test was used. Nine commercial batches of house-dust allergen were studied by means of this test. The results of the determination of the activity of house-dust allergen in the passive hemagglutination inhibition test, the skin tests and the indirect mast-cell degranulation test were in good correlation.     

An indirect ELISA of classical swine fever virus based on quadruple antigenic epitope peptide expressed in <Emphasis Type="Italic">E.coli</Emphasis>

In this study, a synthesized quadruple antigenic epitope gene region of the classical swine fever virus (CSFV) E2 glycoprotein was expressed in E. coli to a obtain target protein. This target protein was used as a coating antigen to establish an indirect ELISA for specifically detecting anti-CSFV antibodies in serum samples from pigs. The P/N cut-off value of this assay was 1.92 by receiver operating characteristic curve (ROC) analysis based on 30 negative sera and 80 positive samples. The test gave 97.5% sensitivity and 96.7% specificity compared with the indirect hemagglutination (IHA) test. The inter-assay and intra-assay coefficients of variation (CVs) for 16 sera were both ⩽6.8%. No cross-reactivity between the coating antigen and anti-bovine viral diarrhoea virus (BVDV) antibodies was observed.     

Serological diagnostics of imported malaria in Czechoslovakia

Two serologic techniques for malaria detection were compared in this study; the indirect fluorescent antibody (IFA) test used in 214 persons (38 Czechoslovak citizens returning from visits to tropical countries and 176 foreign visitors arriving to Czechoslovakia from areas endemic for malaria) and the indirect hemagglutination (IHA) test employed in 125 persons (29 Czechoslovak citizens and 96 foreigners). Comparisons revealed poor correlation between the IFA test and IHA test data. Of the two tests the IFA test appeared to be distinctly more reliable, more sensitive and more specific, the IHA test turned out to yield both false positive and false negative results. The antigen from Plasmodium gallinaceum gave lower IFA titres than P. falciparum antigen, but reacted with antibodies to all species of human plasmodia, and gave reliable test results. Positive serologic responses were appreciably more frequent in foreigners (46.0%) than Czechoslovak citizens (23.7%). The maximum percent positivity for malarial antibody was among individuals from tropical countries of Africa (74.6%), seropositivity in people from malaria endemic areas in Asia and Latin America was far less frequent (28.4% and 44.4%, respectively).