Indian Journal of Animal Research

  • Chief EditorK.M.L. Pathak

  • Print ISSN 0367-6722

  • Online ISSN 0976-0555

  • NAAS Rating 6.43

  • SJR 0.263

  • Impact Factor 0.5 (2023)

Frequency :
Monthly (January, February, March, April, May, June, July, August, September, October, November and December)
Indexing Services :
Science Citation Index Expanded, BIOSIS Preview, ISI Citation Index, Biological Abstracts, Scopus, AGRICOLA, Google Scholar, CrossRef, CAB Abstracting Journals, Chemical Abstracts, Indian Science Abstracts, EBSCO Indexing Services, Index Copernicus
Submit

Research Article

Indian Journal of Animal Research, volume 56 issue 5 (may 2022) : 633-636

RT-PCR Assay for Detection of Enterotropic Strains of Mouse Hepatitis Virus in Faecal Samples

M.R. Srinivasan, K. Vijay, A.K. Karuppannan, S. Ramesh, Y. Krishnamohan Reddy
1Centre for Animal Health Studies, Tamil Nadu Veterinary and Animal Sciences University, Chennai-600 051, Tamil Nadu, India.
Cite article:- Srinivasan M.R., Vijay K., Karuppannan A.K., Ramesh S., Reddy Krishnamohan Y. (2022). RT-PCR Assay for Detection of Enterotropic Strains of Mouse Hepatitis Virus in Faecal Samples. Indian Journal of Animal Research. 56(5): 633-636. doi: 10.18805/IJAR.B-4257.

ABSTRACT

Background: Mouse Hepatitis Virus (MHV) is one of the most important and common viral infections of laboratory mice, due to its highly contagious and subclinical nature, posing threat to the research outcomes. Periodic screening of laboratory mice for MHV is mandatory. Hence, this study was intended to develop sensitive faecal based RT-PCR assays to detect active infection of enterotropic MHV in laboratory mice. 
Methods: Primers targeting N-gene of MHV were selected and their sensitivity was analysed in tenfold serially diluted gene template in the presence of negative mouse faecal cDNA. Thirty-six weaned mice at the age of 6 to 18 weeks were randomly selected at different periods and the blood and faecal sample were collected for serology and RT-PCR assay respectively. RT-PCR assay in colon samples was carried out for comparison. 
Result: PCR assay of MHV detected as low as 4 fg of plasmid DNA. Seroprevalence of MHV is very high than the prevalence by RT-PCR assay showing its retrospective nature and also seroprevalence includes both enterotropic and polytropic strain. RT-PCR results in faecal samples are analogous with that of the colon samples, showing the reliability of antemortem testing of mice for enterotropic strain of MHV.

REFERENCES

  1. Bauer, B.A. and Riley, L.K. (2006). Antemortem detection of mouse parvovirus and mice minute virus by polymerase chain reaction (PCR) of faecal samples. Lab Anim. 40: 144-152.
  2. Beckwith, C.S., Franklin, C.L., Hook, R.R., Besch-willford, C.L. and Riley, L.K. (1997). Faecal PCR assay for diagnosis of Helicobacter infection in laboratory rodents. J. clin. Microbiol. 35(6): 1620-1623.
  3. Casebolt, D.B., Qian, B. and Stephenso, C.B. (1992). Detection of enterotropic mouse hepatitis virus in fecal excretion by polymerase chain reaction. Laboratory animal science. 47(1): 6-10.
  4. Elizabeth, F.M., Rasmussen, L., Fung, P., Amanda, M.A., Luisana, A., David, A.L., Morgan, E.Q., Tammy, D.D., Gloria, M.D. F. and Bianca, A.V. (2012). Prevalence of viral, bacterial and parasitological diseases in rats and mice used in research environments in Australia over a5-y period. Lab Animal Asia Pacific. 3: 4-14.
  5. Kohale, K.N. and Raut, C.G. (2013). Sero-monitoring of common pathogens in various species of laboratory animals at the animal facilities of two research institutes. Indian J. Compo Microbiol. Immunol. Infect. Dis. 34(2): 44-50.
  6. Kundave, V.R., Ram, H., Rafiqi, S.I., Garg, R., Tiwari, A.K. and Benerjee, P.S. (2017). Comparative evaluation of microscopy and PCR assay for detection of Theileria Annulata infections in Ruminants. Journal of Animal Research. 7(4): 699-703.
  7. Nicklas, W., Homberger, F.R., Brunhilde, I.W., Jacobi, K., Kraft, V. and Kunstyr, I. (1999). Implications of infectious agents on results of animal experiment. Lab Anim. 33(1): 39-87.
  8. Pritchett-Corning, K.R., Cosentino, J. and Clifford, C.B. (2009). Contemporary prevalence of infectious agentsin laboratory mice and rats. Lab Anim. 43(2): 165-173. 
  9. Pullium, J.K., Homberger, F.R., Benjamin, K.A., Dillehay, D.L. and Huerkamp, M.J. (2003). Confirmed persistent mouse hepatitis virus infection and transmission by mice with a targeted null mutation of tumour necrosis factor to sentinel mice, Using short-term exposure. Comparitive medicine. 53(4): 439-443.
  10. Scavizzi, F. and Raspa, M. (2004). 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. Lab Anim. 38: 189-199.
  11. Homberger, F.R., Zhang, L. and Barthold, S.W. (1998). Prevalence of Enterotropic and Polytropic Mouse Hepatitis Virus in Enzootically Infected Mouse Colonies. Lab Anim Sci. 48(1): 50-54.
  12. Wan, C.H., Bauer, B.A., Pintel, D.J. and Riley, L.K. (2006). Detection of rat parvovirus type 1 and rat minute virus type 1 by polymerase chain reaction. Lab Anim. 40(1): 63-69.
  13. Wang, R.F., Campbell, W.L., Cao, W.W., Colvert, R.M., Holland, M.A. and Cerniglia, C.E. (1999). Diagnosis of mouse hepatitis virus contamination in mouse population by using nude mice and RT-PCR. Mol cell Probes. 13(1): 29-33.
  14. Yamada, Y.K., Yabe, M., Takimatoo, K., Nakayama, K. and Saitoh, M. (1998). Application of Nested Polymerase chain reaction to detection of Mouse Hepatitis Virus in Fecal Specimens during Natural Outbreak in an Immunodeficient Mouse Colony. Exp. Anim. 47(4): 261-264.
In this Article
    Contact us
    Follow us

    Editorial Board

    View all (0)