In situ detection of hepatitis A virus in cell cultures and shellfish tissues.
An in situ transcription method was developed to detect hepatitis A virus RNA in both cell cultures and shellfish tissues. Radiolabeled cDNA copies were synthesized in situ by reverse transcriptase-directed transcription after annealing with a specific primer to the viral RNA. Both tritium (3H) and 35S were useful in the in situ transcription reaction, but the use of 3H resulted in a lower background and finer detail in the localization of viral particles. Application of the method to different organs of oysters which had bioaccumulated hepatitis A virus allowed the first in situ localization of the virus, specifically in stomach and hepatopancreatic tissues.
Romalde
J L
JL
Division of Molecular Virology, Baylor College of Medicine, Houston, Texas 77030.
Estes
M K
MK
Szücs
G
G
Atmar
R L
RL
Woodley
C M
CM
Metcalf
T G
TG
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D000882:Haplorhini; D006507:Hepatovirus / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D010125:Ostreidae / Q000382:microbiology; D012758:Shellfish / Q000382:microbiology*; D014158:Transcription, Genetic