Anti-HAV ELISA
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产品英文名称:Anti-HAV ELISA
产品中文名称:人的甲型肝炎病毒(HAV)ELISA检测试剂盒
甲型肝炎病毒(HAV)相关介绍:
HAV是一种单链RNA病毒,感染人类肝脏可以引发病毒性甲型肝炎急性传染病,通常通过粪—口途径传播。人与人之间的传播通过摄取被HAV污染的水和食物或直接接触感染者。感染HAV病毒后一般2~6周表现出症状,平均潜伏期在28天。甲型肝炎病毒多侵犯儿童及青年,发病率随年龄增长而递减,40岁上成人中,80%左右均有抗HAV抗体。在甲型肝炎的显性感染或隐性感染过程中,机体都可产生抗HAV的lgM 和lgG抗体。前者在急性期和恢复期出现,后者在恢复后期出现,并可维持多年,对同型病毒的再感染有免疫力。
Anti-HAV ELISA产品使用:
§ 检测原理:Mediagnost anti-HAV EIA, E10 是伪竞争酶联免疫分析。血清或血浆样本添加到之前已涂有一层HAV灭活抗原的微量滴定板小孔中,在 37℃温育2小时。Anti-HAV抗体可以与抗原结合。然后继续添加结合物(过氧化物酶标记的anti-HAV IgG),37℃温育1小时。抗原自由结合位点可以与结合物,冲洗掉多余的结合物,添加底物,在室温下温育30分钟,结合到灭活抗原的结合物会使底物变为蓝色。最后用终止液终止反应,颜色变为黄色。可以在微量滴定板读数器上读出显色反应产物的吸光率。吸光率与anti-HAV滴定量成反比例。定量测定使用标准血清。也可以使用标准试剂制作滴定曲线。
§ 检测过程:A1/A2孔作空白对照→每孔加50µl稀释结合物(除了A1/A2),不要清空和清洗滴定板→反应室密封,37℃温育1小时→清空小孔,用清洗缓冲液清洗三次(300µl/孔)→每孔加100µl底物溶液,A1/A2孔也加,暗处室温(20 – 25℃)温育30分钟→每孔加100µl反应终止液,包括A1/A2孔→以TMB空白底物(A1/A2孔)为准调节分光光度计到450nm,测定每种样本在450nm的分光光度值
§ 结果分析:计算阴性对照和样本的平均分光光度值,阳性对照和阴性对照平均分光光度值误差小于0.4,否则必需重新检测。定点值计算:(阳性对照分光光度值+阴性对照平均分光光度值)/2。如果阳性对照分光光度值小于定点值则认为是阳性。样本平均分光光度值大于定点值则被认为是阴性。
Anti-HAV ELISA英文简述:
Positive detection of antibodies directed against the Hepatitis A virus (anti-HAV) is evidence of immunity to Hepatitis-A virus (1). After natural infection with the Hepatitis A-Virus, neutralising antibodies appear at the same time of Anti-HAV of IgG-Class formation.
Mediagnost anti-HAV EIA, E10 is a pseudo-competitive enzyme immunoassay. Serum or plasma samples are added to the wells of a microtiter plate, which have been previously coated with inactivated HAV antigen, and incubated for 2 hours at 37 °C. Anti-HAV antibodies bind to the antigen. The conjugate (peroxidase labeled anti-HAV IgG) is added and incubated again for 1 h at 37 °C. Free binding sites of the antigen are bound with conjugate. Excess conjugate is washed of the plate and the substrate is added and incubated for 30 min at room temperature. The bound conjugate changes the colour of the subtrate to blue. The reaction is terminated by adding the stopping solution. The colour turns yellow. The absorbance of the coloured reaction product is measured on a microtiter plate reader. The extinction is reciprocal to the anti-HAV titer. For quantitative determination use the included serum standards.
The preparation of titration curve e.g. for calibration of sera by means of standard reagents is also possible.
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