REOVIRUS-3 NOT DETECTED BY REVERSE TRANSCRIPTASE-MEDIATED POLYMERASE CHAIN-REACTION ANALYSIS OF PRESERVED TISSUE FROM INFANTS WITH CHOLESTATIC LIVER-DISEASE

Reovirus type 3 has been implicated in the origin and pathogenesis of extrahepatic biliary atresia and idiopathic neonatal hepatitis, but ro utine detection of this virus in hepatobiliary tissues from affected i nfants by culture and histological techniques has been unsuccessful. I n this study, oligonucleotide primers specific to the M3 genome segmen t of reovirus 3 (Dearing) were used in a reverse transcriptase-mediate d polymerase chain reaction technique to develop a sensitive and speci fic assay for the detection of reovirus 3 RNA in formalin-fixed, paraf fin-embedded patient samples, Optimal reaction conditions were determi ned by testing infected murine tissues and preserved human liver tissu e supplemented; with reovirus 3. Archival specimens from 50 infants, i ncluding 14 with extrahepatic biliary atresia, 20 with idiopathic neon atal hepatitis, and 16 age-matched controls, were evaluated. Successfu l amplification of human albumin complementary DNA from the preserved tissues confirmed the presence of intact RNA in every patient specimen tested. Analysis of the amplification reactions by agarose gel electr ophoresis and Southern blot hybridization detected the presence of reo viral RNA only once in a single patient sample. These results do not s upport a strong role for reovirus 3 in the development of neonatal cho lestatic liver disease, The recent association of other RNA viruses of the Reoviridae family with murine liver disease and human extrahepati c biliary atresia indicates that continued investigation into a viral cause for idiopathic neonatal hepatobiliary disease is warranted.