Reovirus 3 not detected by reverse transcriptase—mediated polymerase chain reaction analysis of preserved tissue from infants with cholestatic liver disease

Abstract

Reovirus type 3 has been implicated in the origin and pathogenesis of extrahepatic biliary atresia and idiopathic neonatal hepatitis, but routine detection of this virus in hepatobiliary tissues from affected infants by culture and histological techniques has been unsuccessful. In this study, oligonucleotide primers specific to the M3 genome segment of reovirus 3 (Dearing) were used in a reverse transcriptase-mediated polymerase chain reaction technique to develop a sensitive and specific assay for the detection of reovirus 3 RNA in formalin-fixed, paraffin-embedded patient samples. Optimal reaction conditions were determined by testing infected murine tissues and preserved human liver tissue supplemented with reovirus 3. Archival specimens from 50 infants, including 14 with extrahepatic biliary atresia, 20 with idiopathic neonatal hepatitis, and 16 age-matched controls, were evaluated. Successful 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 electrophoresis and Southern blot hybridization detected the presence of reoviral RNA only once in a single patient sample. These results do not support a strong role for reovirus 3 in the development of neonatal cholestatic liver disease. The recent association of other RNA viruses of the Reoviridae family with murine liver disease and human extrahepatic biliary atresia indicates that continued investigation into a viral cause for idiopathic neonatal hepatobiliary disease is warranted.

Nonvisualization of the fetal gallbladder by second‐trimester ultrasound scan: strategy of clinical management based on four examples

“…The etiology of biliary duct atresia is unknown, although various hypotheses have been formulated, such as persistence of fetal bile ducts, which release bile into the hepatic parenchyma, leading to an inflammatory reaction that causes fibrosis (Tan et al, 1994). A viral cause has been posited, but the data are contradictory (Steele et al, 1995), and familial cases suggest that a genetic component may be implicated (Lachaux et al, 1988), but no gene has yet been identified.…”

Armed CD4+ Th1 effector cells and activated macrophages participate in bile duct injury in murine biliary atresia

“…Tyler et al [7] reported finding nested RT-PCR evidence of reovirus in infants with BA and Riepenhoff-Talty et al [8] found RT-PCR evidence of group C rotavirus (Reovirdae family) in BA hepatobiliary tissue. However, other investigators failed to find presence of these viruses in BA patients [9,10]. It has recently been proposed that the perinatal/acquired form may be caused by a biliary trophic viral infection, leading to an initial bile duct epithelial injury that triggers a persistent immune-mediated sclerosing process resulting in obstruction of extrahepatic bile ducts [4].…”

Biliary Atresia: A Multidisciplinary Approach to Diagnosis and Management

“…43 These results, however, have not been confirmed by other investigators. 49,50 Similarly, a murine model of biliary atresia induced by group A rotavirus is well described and closely mimics human disease. 46 Interestingly, transfer of T cells from rotavirus-induced biliary atresia to severe combined immunodeficiency mice has been shown to be sufficient to cause bile duct-specific inflammation.…”