应用致敏SPA菌体提高CAV/CPV PCR检出率的研究

为消除粪便等病料中的PCR干扰物质 ,提高犬腺病毒与细小病毒 (CAV CPV)联合PCR检出率。根据免疫吸附的原理 ,应用CAV CPV高免血清致敏含A蛋白的金黄色葡萄球菌 ,制成致敏SPA菌体免疫吸附剂 ;以此免疫吸附提取粪便等病料中的CAV/CPV ,然后直接或经 3 5mol/LMgCl2 将所吸附病毒洗脱后再做PCR。结果 8份经电镜负染或HA/HI试验验证为阳性的粪便样品 ,如此处理后进行PCR检测 ,结果均为阳性 ;而直接取粪便离心后用上清进行PCR检测 ,结果均为阴性。表明该法可有效去除待检样品中的PCR干扰物质 ,提高PCR的检出率。

Improvement of PCR detection rate of CAV/CPV in feces using sensitized Staphylococcus aureus

In order to eliminate inhibitory substances, which interfere the sensitivity of multi polymerase chain reaction (PCR) to CAV CPV from animal fecal specimens, CAV/CPV were adhered to the anti serum sensitized Staphylococcus aureus, and were detected by PCR directly or after elution with 3 5 mol/L MgCl 2. Eight fecal specimens which had been identified to be CAV/CPV positive by electron microscope or HA/HI could not be detected by direct PCR, but after being purified by the method established above, all the specimens were PCR positive. The results indicated that the method was effective to remove the inhibitors and could improve PCR detection rate of the fecal spectimens.