Application of Nested Polymerase Chain Reaction to Detection of Mouse Hepatitis Virus in Fecal. Specimens during a Natural Outbreak in an Immunodeficient Mouse Colony.

Abstract

The usefulness of RT-PCR for the detection of MHV in tissues and feces of experimentally infected animals has been reported, but it was unclear whether the method was also applicable for the detection of MHV during a natural outbreak. Enterotropic infection is considered to be the most common form of natural infection among various forms of MHV infection. In this paper, RT-nested PCR was performed to detect MHV excreted in the feces during an outbreak in an immunocompromised A/WySnJ mouse colony. The expected bands were amplified after nested PCR from 20 fecal samples out of 37. These results showed that RT-nested PCR could be applicable for the diagnosis for MHV natural infection.

Indirect ELISA and Indirect Immunofluorescent Antibody Assay for Detecting the Antibody against Murine Norovirus S7 in Mice

“…Serological tests are the most common methodology used in viral surveillance of laboratory mouse colonies. They include the complement fixation test, hemagglutination inhibition test, neutralization test, [4,21]. An important caveat is that serology is invalid for mice that have just been infected and immunodeficient mice, which cannot produce immunoglobulins.…”

Sequence Analysis of Major Structural Proteins of Newly Isolated Mouse Hepatitis Virus.

“…We also kept A/WySnJ mice with severe B cell deficiency in the room in which an MHV-TY outbreak occurred. The MHV virus gene was detected in the feces of 20 A/WySnJ mice out of 37 [25]. Although pathological changes, focal necrosis and chronic hepatitis were observed in the liver of some A/WySnJ mice, the MHV virus was not detected in the liver.…”

“…MHV primarily infects the respiratory system or the gastrointestinal tract and causes diverse diseases such as hepatitis, encephalomyelitis and enteritis depending on the strain of the virus and the status of the mice [19,22]. Enterotropic infection is considered to be the most common form of natural infection [3].In one animal room in our facility there has recently been an epidemic of MHV [25]. We isolated an epidemic strain of MHV from a fecal pellet in the colon of a BALB/c mouse with X-linked immunodeficiency (xid) housed in this room, and nucleotide sequences of the major structural proteins of this strain were analyzed.…”

“…In one animal room in our facility there has recently been an epidemic of MHV [25]. We isolated an epidemic strain of MHV from a fecal pellet in the colon of a BALB/c mouse with X-linked immunodeficiency (xid) housed in this room, and nucleotide sequences of the major structural proteins of this strain were analyzed.…”

Tissue distribution and duration of mouse hepatitis virus in naturally infected immunocompetent ICR (CD-1) and immunodeficient athymic nude-nu mouse strains used for ovarian transplantation and in vitro fertilization

“…However, molecular detection of the virus with RT-PCR is rapid, more sensitive and allows the detection of the early phases of the infection (Homberger et al 1991, Kunita et al 1992, Yamada et al 1993, Casebolt et al 1997, Wang et al 1999. A further enhanced sensitivity of this method is achieved by the use of nested primers (Matthaei et al 1998, Yamada et al 1998. Colony rederivation methods such as hysterectomy-derivation (HD), embryo-transfer (ET) and IVF have proven successful in eradicating the virus from infected colonies (Reetz et al 1988, Suzuki et al 1996.…”