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通过聚合酶链反应/酶免疫测定法检测胆道闭锁患儿肝胆样本中轮状病毒,未获相关证据。

Lack of evidence for rotavirus by polymerase chain reaction/enzyme immunoassay of hepatobiliary samples from children with biliary atresia.

作者信息

Bobo L, Ojeh C, Chiu D, Machado A, Colombani P, Schwarz K

机构信息

Department of Pediatrics, Johns Hopkins Medical Institutions, Baltimore, Maryland 21287, USA.

出版信息

Pediatr Res. 1997 Feb;41(2):229-34. doi: 10.1203/00006450-199702000-00013.

Abstract

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.

摘要

本研究旨在分析胆道闭锁患者的肝胆样本中A、B和C组轮状病毒的情况。因为A组轮状病毒已被用于建立该疾病的动物模型,且在一组患者的肝胆样本中发现了C组轮状病毒。使用非同位素逆转录酶聚合酶链反应酶免疫测定法,对10例胆道闭锁患者以及14例患有其他肝脏疾病的对照患者的胆管残端和肝组织进行A、B和C组轮状病毒检测。胆道闭锁患者的中位年龄为3个月,且来自特定地理区域。将A组和C组的轮状病毒毒株用作聚合酶链反应阳性对照。这两组轮状病毒RNA的检测限分别为5个空斑形成单位和50个组织培养感染剂量(ID50)。对于A组和C组,组织培养的敏感性比聚合酶链反应低100倍。通过酶免疫测定法或Southern印迹杂交法确定,巢式非同位素探针仅在溶液中与同源靶DNA杂交。尽管在所有肝胆样本中均能检测到β-肌动蛋白管家基因的mRNA,但在胆道闭锁组或阴性对照组中均未发现轮状病毒RNA的证据。总之,轮状病毒不是胆道闭锁常见的病毒病因。

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