Ticehurst J R, Feinstone S M, Chestnut T, Tassopoulos N C, Popper H, Purcell R H
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
J Clin Microbiol. 1987 Oct;25(10):1822-9. doi: 10.1128/jcm.25.10.1822-1829.1987.
Hepatitis A virus (HAV) RNA was extracted from cell culture, serum, liver, and feces and then detected by molecular hybridization with cloned HAV cDNA. Hybridization was approximately 10-fold more sensitive than immune electron microscopy or radioimmunoassay was and less sensitive than was assays of HAV infectivity in primates or in cell culture. As little as 10(3) 50% infective doses of HAV, or approximately 0.1 pg of viral RNA, was detected by this method. Analysis of fecal specimens from an experimentally infected marmoset and an epidemic of hepatitis A showed that HAV excretion could often be detected later in the illness by hybridization than by radioimmunoassay. This technique should be widely applicable for detection and analysis of HAV RNA.
从细胞培养物、血清、肝脏和粪便中提取甲型肝炎病毒（HAV）RNA，然后通过与克隆的HAV cDNA进行分子杂交来检测。杂交的灵敏度比免疫电子显微镜或放射免疫测定法高约10倍，但比在灵长类动物或细胞培养中检测HAV感染性的测定法灵敏度低。用这种方法可检测到低至10³个50%感染剂量的HAV，或约0.1 pg的病毒RNA。对一只实验感染的狨猴的粪便标本以及一次甲型肝炎流行的分析表明，通过杂交检测到HAV排泄的时间往往比放射免疫测定法晚。这项技术应广泛应用于HAV RNA的检测和分析。
