Hostname: page-component-8448b6f56d-dnltx Total loading time: 0 Render date: 2024-04-24T21:34:10.056Z Has data issue: false hasContentIssue false

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

Published online by Cambridge University Press:  19 October 2009

J. U. Umoh
Affiliation:
Departments of Veterinary Public Health and Preventive Medicine, Ahmadu Bello University, Zaria, Kaduna State, Nigeria
C.D. Ezeokoli
Affiliation:
Departments of Surgery and Medicine, Ahmadu Bello University, Zaria, Kaduna State, Nigeria
A. E. J. Okoh
Affiliation:
National Veterinary Research Institute, Vom, Plateau State, Nigeria
Rights & Permissions [Opens in a new window]

Summary

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

Formalin-fixed samples from 221 animal brains received for rabies diagnosis in Nigeria were digested in 0·1% trypsin in phosphate buffered saline, pH 7·4, and smears stained for rabies antigen by direct immunofluorescence (IF). The results were compared with those obtained using fresh material from the same animals for Negri body staining, mouse inoculation (MI) and occasionally immunofluorescent staining.

From 191 specimens examined for Negri bodies and by mouse inoculation 51 and 64 respectively proved positive. The IF smear technique under investigation failed to detect 5 of these but showed up as positive 30 which had been recorded as Negri-negative and 19 that had gone undetected by MI too. In a direct comparison with IF staining of fresh tissue from 23 known rabies positive animals the similar staining of trypsin-digested formalized smears failed to give a positive result in 2 out of 23 cases.

Some problems were encountered with smears not sticking to slides. When gelatinized slides were used fluorescence was sometimes poorer.

Where transport and refrigeration are difficult and section-cutting equipment is lacking the technique of IF staining of smears prepared from formalized brain tissue after treatment with trypsin can be a useful adjunct to other diagnostic methods. It also makes for safer working where special facilities are absent.

Type
Research Article
Copyright
Copyright © Cambridge University Press 1985

References

REFERENCES

Dean, D. J. & Abelseth, M. K. (1973). The fluorescent antibody test. In Laboratory Techniques in Rabies, 3rd ed., pp. 7384. World Health Organization, Geneva.Google ScholarPubMed
Dupont, J. R. & Earle, K. M. (1965). Human rabies encephalitis. A study of forty-nine fatal cases with a review of the literature. Neurology 15, 10231034.CrossRefGoogle ScholarPubMed
Fisohman, H. R. & Ward, F. E. (1965). Infectivity of fixed impression smears prepared from rabies virus-infected brain. American Journal of Veterinary Research 30, 22052208.Google Scholar
Huang, S.-N., Minassian, H. & More, J. D. (1970). Application of immunofluorescent staining on paraffin sections improved by trypsin digestion. Laboratory Investigation 35, 383390.Google Scholar
Johnson, K. P., Swoveland, P. T. & Emmons, R. W. (1980). Diagnosis of rabies by immunofluorescence in trypsin-treated histologic sections. Journal of American Medical Association 244, 4143.CrossRefGoogle ScholarPubMed
Koprowski, H. (1973). The mouse inoculation test. In Laboratory Techniques in Rabies, 3rd ed. (ed. Kaplan, M. M. and Koprowski, H.), pp. 8593. World Health Organization, Geneva.Google ScholarPubMed
Lewis, V. J. & Thacker, W. L. (1974). Limitations of deteriorated tissue for rabies diagnosis. Health Laboratory Science 11, 812.Google ScholarPubMed
Reid, F. L., Hall, N. H., Smith, J. S. & Baer, G. M. (1983). Increased immunofluorescent staining of rabies-infected, formalin-fixed brain tissue after pepsin and trypsin digestion. Journal of Clinical Microbiology 18, 968971.CrossRefGoogle ScholarPubMed
Rowse-Eagle, D., Watson, H. D. & Tignor, G. H. (1981). Improved method for trypsin digestion of paraplast sections before immunofluorescence staining. Journal of Clinical Microbiology 13, 996997.CrossRefGoogle ScholarPubMed
Umoh, J. U. & Blenden, D. C. (1981). Immunofluorescent staining of rabies virus antigen in formalin-fixed tissue after treatment with trypsin. Bulletin of the World Health Organization 59, 737744.Google ScholarPubMed