Lack of evidence for rotavirus by polymerase chain reaction/enzyme immunoassay of hepatobiliary samples from children with biliary atresia.
The purpose of this study was to analyze hepatobiliary samples of patients with biliary atresia for rotavirus groups. A, B, and C, because group A rotavirus had been used to produce an animal model of the disease and group C rotavirus had been found in hepatobiliary samples from one group of patients. Biliary remnants and liver tissue from 10 biliary atresia and 14 control patients with other liver diseases were examined for rotavirus groups A, B, and C using nonisotopic, reverse transcriptase polymerase chain reaction enzyme immunoassay. Biliary atresia patients had a median age of 3 mo and were from a confined geographic area. Rotaviral stocks from groups A and C were used as polymerase chain reaction-positive controls. The limits of detection for rotaviral RNA from these two groups were respectively, 5 plaque-forming units and 50 tissue culture infectious doses (ID50). Tissue culture was 100-fold less sensitive for groups A and C than the polymerase chain reaction. The nested nonisotopic probes hybridized in solution only with their homologous target DNAs as determined by the enzyme immunoassay, or by Southern blot hybridization. Although it was possible to detect mRNA from a beta-actin housekeeping gene in all of the hepatobiliary samples, no evidence of rotaviral RNA was found in either the biliary atresia or the negative control group. In conclusion, rotavirus is not a common viral etiology of biliary atresia.
Bobo
L
L
Department of Pediatrics, Johns Hopkins Medical Institutions, Baltimore, Maryland 21287, USA.
Ojeh
C
C
Chiu
D
D
Machado
A
A
Colombani
P
P
Schwarz
K
K
D000293:Adolescent; D001652:Bile Ducts / Q000821:virology*; D001656:Biliary Atresia / Q000821:virology*; D002648:Child; D005260:Female; D006801:Humans; D007124:Immunoenzyme Techniques; D007223:Infant; D008099:Liver / Q000821:virology*; D008297:Male; D016133:Polymerase Chain Reaction; D012401:Rotavirus / Q000302:isolation & purification*; D012680:Sensitivity and Specificity