Sensitivity comparison of immunofluorescent techniques for detection of rabies virus antigen from brain specimens

A B BEIGH; B S SANDHU; C K SINGH; K GUPTA; N K SOOD

doi:10.56093/ijans.v84i2.37834

Authors

A B BEIGH Sher-e-Kashmir University of Agricultural Sciences and Technology of Jammu, R S Pura, Jammu and Kashmir 181 102 India
B S SANDHU Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab 141 004 India
C K SINGH Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab 141 004 India
K GUPTA Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab 141 004 India
N K SOOD Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab 141 004 India

https://doi.org/10.56093/ijans.v84i2.37834

Keywords:

Animals, FAT, Formalin fixed tissue, Impression smear, Rabies

Abstract

The fluorescent antibody test (FAT) is most preferred method to visualize fine particles of rabies viral antigen in rabies infected tissues. In present study, formalin-fixed brain tissues from 100 clinically rabid animals were heat treated and tested for the presence of rabies virus antigen by direct immunofluorescent (IF) staining. The results were compared with those obtained from direct IF staining of acetone- fixed smears. The evaluation of rabies status between fresh and formalin – fixed tissue was in agreement in 100% of the cases. These results suggested that this technique is a valid procedure for routine diagnosis of rabies, particularly in situations in which fresh tissue is not available.

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References

Abreu C C, Nakayama P A, Nogueira C I, Mesquita L P, Lopes P F, Varaschin M S, Seixas J N, Ferreira E and Bezerra P S Jr. 2012. Domestic microwave processing for rapid immuno- histochemical diagnosis of bovine rabies. Histology and Histopathology 27(9): 1227–30.

Awahan S, Sandhu B S, Singh C K, Gupta K, Sood N K and Kaw A. 2012. Comparison of clinico-pathological, immuno- histochemical and immunofluorescent techniques for diagnosis of rabies in animals. Indian Journal of Animal Sciences 82(1): 3–8.

Bourhy H and Sureau P. 1990. Laboratory Methods for Rabies Diagnosis. Collection of Institute Louis Pasteur. pp.154 –197. Paris, France.

Goldwasser R A and Kissling R E. 1958. Fluorescent antibody staining of street and fixed rabies virus antigen. Proceedings of the Society for Experimental Biology and Medicine 98: 291– 323.

Gunawardena G S P de S and Blackemore W F. 2007. Immuno- histochemical detection of rabies virus antigen in the brainstem and spinal cord of rabid dogs in Sri Lanka. Proceedings of the Peradeniya University Research Sessions 12(1): 168.

Johnson K P, Swoveland P T and Emmons R W. 1980. Diagnosis of rabies by immunofluorescencein trypsin-treated histologic sections. Journal of American Medical Association 244: 41– 43.

Perrin P and Sureau P. 1987. A collaborative study of an experimental kit for rapid rabies enzyme immunodiagnosis. Bulletin WHO 65(4): 489–93.

Reid F L, Hall N H, Smith J S and Baer G M. 1983. Increased immunofluorescent staining of rabies-infected, formalin-fixed brain tissue after pepsin and trypsin digestion. Journal of Clinical Microbiology 18: 968–71.

Smith J. 1999. Rabies virus. Manual of Medical Microlbiology. (Eds) Murray, P R, Baron E J, Pfaller M A, Tenover F C and Yolken R H. 7th edn, pp. 1099–106. American Society for Microbiology, Washington D C.

Swoveland P T and Johnson K P. 1979. Enhancement of fluorescent antibody staining of viral antigens in formalin-fixed tissues by trypsin digestion. Journal of Infectious Diseases 140: 758–64.

Umoh J U, Ezeokoli C D and Okoh A E. 1985. Immunofluorescent staining of trypsinized formalin-fixed brain smears for rabies antigen: Results compared with those obtained by standard methods for 221 suspect animal cases in Nigeria. Journal of Hygiene 94: 129–34.

Velleca W M and Forrester F T. 1981. Laboratory Methods for Detecting Rabies. U S Government Printing Office, Washington DC.

Warner C K, Whitfield S G, Fekadu M and HO H. 1997. Procedures for reproducible detection of rabies virus antigen mRNA and genome in situ in formalin-fixed tissues. Journal of Virological Methods 67: 5–12.

Whitfield C G, Fekadu M, Shaddock J H, Niezgoda M, Warner C K and Messenger S L. 2001. A comparative study of the fluorescent antibody test for rabies diagnosis in fresh and formalin-fixed brain tissue specimens. Journal of Virological Methods 95: 145–5.