Use of Polymerase Chain Reaction to Diagnose a Natural Outbreak of Mouse Hepatitis Virus Infection in Nude Mice
Abstract:The enormous cost of eliminating mouse hepatitis virus (MHV) from a mouse colony demands that a confirmed etiologic diagnosis be made to justify the necessary remedial action. We describe an outbreak of MHV in nude mice in which histopathologic findings provided a presumptive diagnosis, but results of serologic testing of affected nude mice and immunocompetent sentinels were negative. Results of trans- mission electron microscopy of liver specimens from affected mice were equivocal. Confirmation of the etio- pathogenesis was eventually provided by reverse transcriptase-polymerase chain reaction (RT-PCR), using primers with nested sequences directed to two separate but highly conserved regions of the MHV genome. This procedure detected MHV in the liver of an affected nude mouse and in a sentinel, although in the latter a positive result was obtained only because of the increased sensitivity of nested primers used in a second round of amplification. Virus was not detected in cell lines that had been injected into the mice, and the source of the outbreak was not found. These results indicate the applicability of RT-PCR for detecting MHV in a field situation while also illustrating that conventional, complementary techniques still have an essential role in reaching a diagnosis. It is recommended that specimens should be taken for histologic examination and serologic testing, as well as for molecular studies when MHV infection is suspected.
Document Type: Research Article
Affiliations: 1: Gene Targeting, Division of Biochemistry and Molecular Biology, The John Curtin School of Medical Research, Australian National University, Canberra, ACT, Australia 2: Oncology Research Centre, Prince of Wales Hospital and Department of Medicine, University of New South Wales, Sydney, NSW, Australia 3: Department of Veterinary Anatomy and Pathology, University of Sydney, NSW, Australia 4: Electron Microscopy Unit, Department of Anatomical Pathology, University of New South Wales, Sydney, NSW, Australia 5: Oncology Research Centre, Prince of Wales Hospital and Department of Medicine, University of New South Wales, Sydney, NSW, Australia, Oncology Research Centre, POWH, Institute of Oncology, Villa 1, Prince of Wales Hospital, High Street, Randwick, NSW 2031, Australia
Publication date: 1998-04-01
Comparative Medicine (CM), an international journal of comparative and experimental medicine, is the leading English-language publication in the field and is ranked by the Science Citation Index in the upper third of all scientific journals. The mission of CM is to disseminate high-quality, peer-reviewed information that expands biomedical knowledge and promotes human and animal health through the study of laboratory animal disease, animal models of disease, and basic biologic mechanisms related to disease in people and animals.
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