Rodák L, Smíd B, Nevoránková Z, Smítalová R, Valícek L
Veterinary Research Institute, Hudcova 70, 621 32 Brno, Czech Republic.
J Vet Med B Infect Dis Vet Public Health. 2004 May;51(4):160-5. doi: 10.1111/j.1439-0450.2004.00746.x.
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.
制备了针对A组轮状病毒Vp6蛋白的单克隆抗体,并用于验证三种用于检测A组轮状病毒的阻断酶联免疫吸附测定(ELISA)改良方法。选用竞争阻断ELISA(CB-ELISA)和电子显微镜(EM)对194份腹泻仔猪的现场粪便样本进行检测。通过CB-ELISA方法在43份样本(22.2%)中检测到轮状病毒,而通过EM检测在26份样本(13.4%)中检测到轮状病毒。然而,在EM检测呈阳性的26份样本中,CB-ELISA在19份样本(73.1%)中检测到A组轮状病毒;这表明A组轮状病毒在所有由轮状病毒引起的胃肠炎病例中的占比。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)的对照样本,以及通过与商业ELISA试剂盒对样本进行比较检测,验证了CB-ELISA的敏感性和特异性。在存在螯合剂的情况下对样本进行检测,对CB-ELISA的敏感性有积极影响。