4.4 Article

Development of a generic real-time PCR assay for simultaneous detection of proviral DNA of simian Betaretrovirus serotypes 1, 2, 3, 4 and 5 and secondary uniplex assays for specific serotype identification

期刊

JOURNAL OF VIROLOGICAL METHODS
卷 162, 期 1-2, 页码 148-154

出版社

ELSEVIER SCIENCE BV
DOI: 10.1016/j.jviromet.2009.07.030

关键词

Simian betaretrovirus; SRV; PCR; SPF

资金

  1. National Center for Research Resources (NCRR) of the National Institutes of Health (NIH) [RR00169]
  2. NATIONAL CENTER FOR RESEARCH RESOURCES [P51RR000169] Funding Source: NIH RePORTER
  3. OFFICE OF THE DIRECTOR, NATIONAL INSTITUTES OF HEALTH [P51OD010425, P51OD011107] Funding Source: NIH RePORTER

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Simian betaretroviruses (formerly Type D retroviruses; SRV) are a group of closely related retroviruses for which the natural host species are Asian monkeys of the genus Macaca. Five serotypes have been identified by classical neutralization assays and three additional untyped variants have been reported (SRVTsukuba, SRV-6, SRV-7). These viruses may be significant pathogens in macaque colonies, causing a broad spectrum of clinical disease secondary to viral-induced immune suppression. Undetected SRV infections in research macaques also represent a potential confounding variable in research protocols and a concern for human caretakers. Intensive testing efforts have been implemented to identify infected animals in established colonies. A real-time quantitative generic multiplex PCR assay was developed that is capable of simultaneous detection of proviral DNA of SRV serotypes 1, 2, 3, 4 and 5. This assay incorporates amplification of the oncostatin M (OSM) gene for confirmation of amplifiable DNA and allows quantitation of the number of proviral copies per cell analyzed in each multiplex reaction. Detection of multiple serotypes by PCR increases the efficiency and cost-effectiveness of SRV screening programs. A panel of SRV serotype-specific uniplex real-time PCR assays for discrimination among the five recognized serotypes is also described. (C) 2009 Elsevier B.V. All rights reserved.

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