应用反向被动血凝试验检测粪便中的牛冠状病毒
Detection of bovine coronavirus in feces by reversed passive hemagglutination.
作者信息
Sato K, Inaba Y, Tokuhisa S, Miura Y, Kaneko N, Asagi M, Matumoto M
出版信息
Arch Virol. 1984;80(1):23-31. doi: 10.1007/BF01315291.
A reversed passive hemagglutination (RPHA) method was developed for the detection of bovine coronavirus in fecal specimens. Sheep erythrocytes fixed with glutaraldehyde, and then treated with tannic acid were coated with anti-bovine coronavirus rabbit antibodies purified by affinity chromatography using bovine coronavirus linked to Sepharose 4B. The RPHA test was carried out by a microtiter method. Erythrocytes coated with purified specific antibodies were agglutinated by bovine coronavirus, but not by bovine rotavirus or enterovirus. The reaction was inhibited by antiserum to bovine coronavirus, confirming the specificity of the reaction. The RPHA test detected bovine coronavirus in 13 of 22 fecal specimens (59 per cent), from natural cases of diarrhea, while the positive rates were only 14 per cent (3/22) and 22 per cent (5/22) for immunofluorescent staining of primary cultures of calf kidney cells infected with the specimens, and immune electron microscopy respectively. The advantages of the RPHA method are its simplicity, high sensitivity and rapidity.
建立了一种反向被动血凝（RPHA）方法用于检测粪便标本中的牛冠状病毒。用戊二醛固定的绵羊红细胞，再经单宁酸处理后，用通过与琼脂糖4B偶联的牛冠状病毒经亲和层析纯化的抗牛冠状病毒兔抗体包被。RPHA试验采用微量滴定法进行。用纯化的特异性抗体包被的红细胞被牛冠状病毒凝集，但不被牛轮状病毒或肠道病毒凝集。该反应被抗牛冠状病毒抗血清抑制，证实了反应的特异性。RPHA试验在22份腹泻自然病例的粪便标本中检测到13份（59%）牛冠状病毒，而对感染这些标本的犊牛肾细胞原代培养物进行免疫荧光染色和免疫电镜检查的阳性率分别仅为14%（3/22）和22%（5/22）。RPHA方法的优点是简便、灵敏度高和快速。
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