用于在生物样本中鉴定小鼠多瘤病毒的诊断性聚合酶链反应检测法。

Diagnostic polymerase chain reaction assays for identification of murine polyomaviruses in biological samples.

作者信息

Carty A J, Franklin C L, Riley L K, Besch-Williford C

机构信息

University Laboratory Animal Resources, University of Pennsylvania, Philadelphia 19104, USA.

出版信息

Comp Med. 2001 Apr;51(2):145-9.

PMID:11922178

Abstract

PURPOSE

Mouse polyoma virus and K virus are murine polyomaviruses frequently used in carcinogenicity and cellular biology studies in mice. These viruses can cause persistent infections, which increase the likelihood of transmission through transplantation of cells from infected mice. To identify polyomavirus-infected biological samples, several diagnostic polymerase chain reaction (PCR) assays were developed.

METHODS

Polyomavirus-family and virus-specific PCR assays were designed and optimized for specificity and sensitivity. The generic (polyomavirus-family) PCR assay and mouse polyoma virus-specific assays were compared with the mouse bioassay for diagnosis of infected cellular samples.

RESULTS

Specificity of the PCR assays was confirmed by testing a battery of other murine viruses. The mouse polyoma virus PCR test was the most sensitive assay, detecting as few as 2,000 copies of homologous virus. The K virus PCR assay was about eightfold less sensitive, and the generic PCR test was the least sensitive. Mouse polyoma virus and generic PCR assays amplified mouse polyoma virus in the inoculum and tissues from experimentally infected mice, and performed better than did the mouse bioassay.

CONCLUSIONS

Results of this study confirm that PCR is a specific and sensitive method for detection of murine polyomaviruses in biological samples.

摘要

目的

小鼠多瘤病毒和K病毒是常用于小鼠致癌性和细胞生物学研究的鼠多瘤病毒。这些病毒可引起持续性感染，这增加了通过移植来自感染小鼠的细胞进行传播的可能性。为了鉴定多瘤病毒感染的生物样本，开发了几种诊断性聚合酶链反应（PCR）检测方法。

方法

设计并优化了多瘤病毒家族和病毒特异性PCR检测方法，以提高特异性和敏感性。将通用（多瘤病毒家族）PCR检测方法和小鼠多瘤病毒特异性检测方法与小鼠生物测定法进行比较，以诊断感染的细胞样本。

结果

通过检测一系列其他鼠病毒证实了PCR检测方法的特异性。小鼠多瘤病毒PCR检测是最灵敏的检测方法，可检测低至2000个同源病毒拷贝。K病毒PCR检测的灵敏度约低八倍，而通用PCR检测最不灵敏。小鼠多瘤病毒和通用PCR检测方法在接种物和实验感染小鼠的组织中扩增了小鼠多瘤病毒，并且比小鼠生物测定法表现更好。