Anti-HAV ELISA
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產品英文名稱:Anti-HAV ELISA
產品中文名稱:人的甲型肝炎病毒(HAV)ELISA檢測試劑盒
甲型肝炎病毒(HAV)相關介紹:
HAV是一種單鏈RNA病毒,感染人類肝臟可以引發(fā)病毒性甲型肝炎急性傳染病,通常通過糞—口途徑傳播。人與人之間的傳播通過攝取被HAV污染的水和食物或直接接觸感染者。感染HAV病毒后一般2~6周表現(xiàn)出癥狀,平均潛伏期在28天。甲型肝炎病毒多侵犯兒童及青年,發(fā)病率隨年齡增長而遞減,40歲上成人中,80%左右均有抗HAV抗體。在甲型肝炎的顯性感染或隱性感染過程中,機體都可產生抗HAV的lgM 和lgG抗體。前者在急性期和恢復期出現(xiàn),后者在恢復后期出現(xiàn),并可維持多年,對同型病毒的再感染有免疫力。
Anti-HAV ELISA產品使用:
§? 檢測原理:Mediagnost anti-HAV EIA, E10 是偽競爭酶聯(lián)免疫分析。血清或血漿樣本添加到之前已涂有一層HAV滅活抗原的微量滴定板小孔中,在 37℃溫育2小時。Anti-HAV抗體可以與抗原結合。然后繼續(xù)添加結合物(過氧化物酶標記的anti-HAV IgG),37℃溫育1小時??乖杂山Y合位點可以與結合物,沖洗掉多余的結合物,添加底物,在室溫下溫育30分鐘,結合到滅活抗原的結合物會使底物變?yōu)樗{色。最后用終止液終止反應,顏色變?yōu)辄S色??梢栽谖⒘康味ò遄x數(shù)器上讀出顯色反應產物的吸光率。吸光率與anti-HAV滴定量成反比例。定量測定使用標準血清。也可以使用標準試劑制作滴定曲線。
§? 檢測過程:A1/A2孔作空白對照→每孔加50μl稀釋結合物(除了A1/A2),不要清空和清洗滴定板→反應室密封,37℃溫育1小時→清空小孔,用清洗緩沖液清洗三次(300μl/孔)→每孔加100μl底物溶液,A1/A2孔也加,暗處室溫(20 – 25℃)溫育30分鐘→每孔加100μl反應終止液,包括A1/A2孔→以TMB空白底物(A1/A2孔)為準調節(jié)分光光度計到450nm,測定每種樣本在450nm的分光光度值
§? 結果分析:計算陰性對照和樣本的平均分光光度值,陽性對照和陰性對照平均分光光度值誤差小于0.4,否則必需重新檢測。定點值計算:(陽性對照分光光度值+陰性對照平均分光光度值)/2。如果陽性對照分光光度值小于定點值則認為是陽性。樣本平均分光光度值大于定點值則被認為是陰性。
Anti-HAV ELISA英文簡述:
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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