聚合酶链反应与病毒分离法在检测自然感染鹿临床样本中流行性出血病病毒的比较

Comparison of polymerase chain reaction and virus isolation for detection of epizootic hemorrhagic disease virus in clinical samples from naturally infected deer.

作者信息

Aradaib I E, Akita G Y, Pearson J E, Osburn B I

机构信息

Department of Veterinary Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis 95616, USA.

出版信息

J Vet Diagn Invest. 1995 Apr;7(2):196-200. doi: 10.1177/104063879500700205.

DOI:10.1177/104063879500700205

PMID:7619901

Abstract

We compared our recently reported reverse transcriptase polymerase chain reaction (PCR)-based assay for detection of epizootic hemorrhagic disease virus (EHDV) in clinical samples with different virus isolation (VI) procedures. Thirty-six blood samples and 1 spleen sample from deer were assessed by the EHDV PCR assay and VI in baby hamster kidney (BHK)-21 cells and embryonated chicken eggs (ECE). The EHDV PCR assay detected EHDV RNA from 6 blood samples obtained from deer during 1988-1989 outbreaks of epizootic hemorrhagic disease and from the spleen and blood samples of a deer with clinical hemorrhagic disease in 1992. The 6 blood samples from the 1988-1989 outbreaks and the spleen sample from the 1992 case were VI positive on BHK-21 cell culture. The blood from the same deer with the PCR- and VI-positive spleen was VI negative in BHK-21 cells and ECE. All EHDV isolates were identified as EHDV serotype 2 by a plaque inhibition test. The results of this study indicate that the sensitivity of the previously described EHDV PCR assay is comparable to or greater than that of the VI method in BHK-21 cell culture or ECE. The EHDV PCR assays could provide a superior diagnostic alternative to the current cumbersome and time-consuming VI procedures.

摘要

我们将我们最近报道的基于逆转录酶聚合酶链反应（PCR）的方法用于检测临床样本中的 epizootic hemorrhagic disease virus（EHDV），并与不同的病毒分离（VI）程序进行了比较。通过 EHDV PCR 检测法以及在幼仓鼠肾（BHK）-21 细胞和鸡胚（ECE）中的 VI 方法，对来自鹿的 36 份血液样本和 1 份脾脏样本进行了评估。EHDV PCR 检测法从 6 份血液样本中检测到了 EHDV RNA，这些样本取自 1988 - 1989 年 epizootic hemorrhagic disease 疫情期间的鹿，以及 1992 年一头患有临床出血性疾病的鹿的脾脏和血液样本。1988 - 1989 年疫情中的 6 份血液样本以及 1992 年病例的脾脏样本在 BHK - 21 细胞培养中 VI 呈阳性。来自同一头 PCR 和 VI 检测均为阳性脾脏的鹿的血液，在 BHK - 21 细胞和 ECE 中 VI 呈阴性。通过蚀斑抑制试验，所有 EHDV 分离株均被鉴定为 EHDV 血清型 2。本研究结果表明，先前描述的 EHDV PCR 检测法的灵敏度与在 BHK - 21 细胞培养或 ECE 中的 VI 方法相当或更高。EHDV PCR 检测法可为当前繁琐且耗时的 VI 程序提供一种更好的诊断替代方法。