Verification of sensitivity and specificity of group a rotavirus detection in piglets faeces with monoclonal blocking ELISA methods
Verification of sensitivity and specificity of group a rotavirus detection in piglets faeces with monoclonal blocking ELISA methods
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DOI：
10.1111/j.1439-0450.2004.00746.x
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发表时间：
2004-01-01
期刊：
影响因子：
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通讯作者：
Valícek, L
中科院分区：
文献类型：
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作者：
Rodák, L;Smíd, B;Valícek, L
制备了抗A组轮状病毒VP6蛋白的单抗，并用3种改进的阻断酶联免疫吸附试验(EL ISA)检测轮状病毒A。用选择性竞争阻断EL ISA(CB-EL ISA)和电子显微镜(EM)检测了194份腹泻仔猪的粪便样品。CB-ELISA法检出轮状病毒43例(22.2%)，EM法检出26例(13.4%)。然而，在EM阳性的26份样品中，有19份(73.1%)用CB-EL ISA法检测到A组轮状病毒，表明A组轮状病毒在所有由轮状病毒引起的胃肠炎病例中所占比例较大。通过在每次分析中加入含有传染性胃肠炎病毒(TGEV)和猪流行性腹泻病毒(PEDV)的对照样品，以及通过与商品化的ELISA试剂盒进行对比检验，验证了CB-ELISA的敏感性和特异性。在有络合剂存在的情况下，样品的检测对CB-EL ISA的灵敏度有积极影响。
Monoclonal antibodies to group A rotavirus Vp6 protein were prepared and used for verification of three blocking enzyme-linked immunosorbent assay (ELISA) modifications to detect rotavirus A. Selected competitive blocking ELISA (CB-ELISA) and electron microscopy (EM) were used for examination of 194 field faecal samples of piglets affected with diarrhoea. Rotavirus was detected in 43 samples (22.2%) by CB-ELISA method, whereas in 26 (13.4%) samples by EM examination. However, of 26 samples positive by EM, rotavirus A was detected by CB-ELISA in 19 (73.1%) samples, indicating the share of group A rotavirus in all cases of gastroenteritis caused by rotavirus. The sensitivity and specificity of the CB-ELISA was verified both by inclusion of control samples containing transmissible gastroenteritis virus (TGEV) and porcine epidemic diarrhoea virus (PEDV) in each analysis and by comparative examination of samples with the commercial ELISA kit. The CB-ELISA sensitivity was positively affected by examination of samples in the presence of chelating agent.