Detection of hepatitis A virus by extraction of viral RNA and molecular hybridization.
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.
Ticehurst
J R
JR
Laboratory of Infectious Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892.
Feinstone
S M
SM
Chestnut
T
T
Tassopoulos
N C
NC
Popper
H
H
Purcell
R H
RH
D000818:Animals; D001345:Autoradiography; D002143:Callitrichinae; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics; D005243:Feces / Q000382:microbiology; D006506:Hepatitis A / Q000175:diagnosis*; D006507:Hepatovirus / Q000235:genetics* / Q000302:isolation & purification; D006801:Humans; D008099:Liver / Q000382:microbiology; D009693:Nucleic Acid Hybridization; D011237:Predictive Value of Tests; D012367:RNA, Viral / Q000032:analysis*; D011863:Radioimmunoassay
D004279:DNA, Viral; D012367:RNA, Viral