Mugale Madhav NİLAKANTH, Bhupinder Singh SANDHU, Beigh AKEEL, Gupta KULDEEP, Sood Naresh KUMAR, Charan Kamal SİNGH

Kedilerde kuduz diagnozunda farklı tekniklerin karşılaştırılması

Amaç: Mevcut araştırmanın amacı kedilerde kuduz teşhisinde en iyi diagnostik metottu belirlemek ve değerlendirmektir. Gereç ve Yöntem: Kuduz süşhesi ile ölmüş 5 kedi değerlendirildi. Şüpheli kedilerin beyinleri toplandı. Direct Fluorescent Antibody Test (dFAT), Hematoxylin & Eosin (H&E) ve Immunohistochemistry (IHC) histopatolojik diagnostik testler uygulandı.Bulgular: Kedilerin salya ve beyinlerine dFAT testi uygulandı ve 4 kedide kuduz virüsü açısından pozitif bulundu. Ölümden sonra Hematoxylin & Eosin (H&E) ve Immunohistochemistry (IHC) teknikleri uygulandı. Vakanın birinde meningitis ve Negri cisimciği gözlendi. Hipokampüs ve serebellumun her bölgesinde sayılan 100 hücrede H&E ve IHC açısından viral antijen varlığı gözlendi. Serebellumun Purkinje hücrelerinin %69’unda ve hipokampüs pramidial hücrelerinin %87’sinde kuduz viral antijeni belirlendi. Öneri: IHC testi dFAT testinin yerine kullanılabilen değerli bir diagnostik testtir. IHC orta dereceli enfeksiyonlar pozitiftir ve retrospektif çalışmalar için oldukça değerlidir. Ayrıca kuduz pozitif beyin örneklerinin taşınmasında halk sağlığı açısından minimum risk taşımaktadır.

Anahtar Kelimeler:

Beyin, direct fluorescent antibody test, histopatoloji, immunohistochemistry, kuduz

Comparison of various techniques used for diagnosis of rabies in cats

Aim: Objectives of this study was to compare and evaluate the best method for diagnosis of rabies in cats. Materials and Methods: Antemortem examination of 5 suspected cats were evaluated. Brains were collected from the suspected cats. Direct Fluorescent Antibody Test (dFAT), Hematoxylin and Eosin (H & E) and Immunohistochemistry (IHC) histopathological diagnostic tests were applied.Results: The dFAT test was conducted on the saliva and fresh brain impression smear of all cats, among that 4 cats showed positive for rabies virus. After death of cats, Hematoxylin & Eosin (H&E) and Immunohistochemistry (IHC) techniques were carried out. One case showed meningitis and remaining showed Negri bodies. Viral antigen depositions were observed by counting 100 cells each region of hippocampus and cerebellum by H&E and IHC. Hippocampus pyramidal cell showed 87% and cerebellum Purkinje cells showed 69% of rabies viral antigen deposition. Conclusion: The IHC can be used as reliable diagnostic technique in addition to dFAT. IHC shows positivity in mildly infected cases and having immense value for retrospective studies. It also minimizes the risk of public health hazard during shipping of rabid positive brain samples.

Keywords:

Brain, direct fluorescent antibody test, histopathology, immunohistochemistry, rabies,

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Bernard KW, 1985. Clinical rabies in humans. In: Rabies concepts for medica professionals, Edition Winkler WG, Merieux Institute, Miami, FL, USA, pp: 45-46.
Birgham J, Van der Merwe M, 2002. Distribution of rabies antigen in infected brain material: determining the reliability of different regions of the brain for the rabies fluorescent antibody test. J Virol Methods, 101, 85-94.
Bourhy H, Rollin PE, Vincent J, Sureau P, 1989. Comparative field evaluation of the fluorescent antibody test, virus isolation from tissue culture, and enzymes immunodiagnosis for rapid laboratory diagnosis of rabies. J Clin Microbiol, 27, 519-523.
Deborah M, Haines E, Clark G, 1991. Enzyme immunohistochemical staining of formalin fixed tissues for diagnosis in veterinary pathology. Can Vet J, 32, 295-202.
Eng TR, Fishbein DB, 1990. Epidemiologic factors, clinical findings and vaccination status of rabies in cats and dogs in the United States in 1988. JAVMA, 197, 201.
Faizee N, Hailat, NQ, Ababneh MM, Hananeh WM, Muhaidat A, 2012. Pathological, immunological and molecular diagnosis of rabies in clinically suspected animals of different species using four detection techniques in Jordan. Transbound Emerg Dis, 59, 154-164.
Feiden W, Kaiser E, Gerhard L, Dahme E, Gylstorff B, Wandeler A, Ehrensberger F, 1988. Immunohistochemical staining of rabies virus antigen with monoclonal and polyclonal antibodies in paraffin tissue sections. J Vet Med, 35, 247-255.
Fekadu M, 1991. Latency and aborted rabies. In: The Natural History of Rabies, Ed: Baer GM, CRC Press, Boca Raton, Florida, USA, pp: 191-198.
Fogelman V, Fischman H, Horman JT, Grigor JK, 1993. Epidemiologic and clinical characteristics of rabies in cats. J Am Vet Med Assoc, 202, 1829- 1838.
Gunawardena GSP de S, Blakemore WF, 2007. Immunohistochemical detection of rabies virus antigen in the brainstem and spinal cord of rabid dogs in Sri Lanka. Proceedings of the Peradeniya University Research Sessions, Sri Lanka, 12, 168-169.
Jogai S, Radotra BD, Banerjee AK, 2001. Immunohistochemical study of human rabies. Neuropathology, 20, 197-203.
Jubb KVE, Huxtable CR, 1996. The nervous system. In: Pathology of Domestic Animals, volume 1, 4th edition, Eds; Jubb KVF, Kennedy PC, Palmer N, Academic Press Inc. Orlando, Florida, USA, pp: 404-05.
Krebs JW, Mark l, Wilson, K, James E, 1995. Rabies-epidemiology, prevention and future research. J Mammal, 76, 681-694.
Last RD, Jardine JE, Smit MM, van der Lugt JJ, 1994. Application of immunoperoxidase techniques to formalin-fixed brain tissue for the diagnosis of rabies in southern Africa. Onderstepoort J Vet Res, 61, 183-187.
Lembo T, Niezgoda M, Velasco-Villa A, Cleaveland S, Ernest E, Rupprecht CE, 2006. Evaluation of a direct, rapid immunohistochemical test for rabies diagnosis. Emerg Infect Dis, 12, 310-13.
Luna LG, 1968. Manual of Histologic Staining Methods, 3rd edition, Armed Forces Institute of Pathology, McGraw-HillToronto, USA, pp: 155-157.
Maxie MG, Youssef S, 2007. Nervous system. In: Jubb, Kennedy, Palmer’s Pathology of Domestic Animals, Ed; Maxie MG, Elsevier, Philadelphia, USA, pp: 281-457.
Meslin M, Koprowski H, Kaplan MM, 1996. Laboratory Techniques in Rabies, 4th edition, WHO, Geneva, Italy, pp: 53-55.
Metze K, Feiden W, 1991. Immunohistochemical detection of rabies virus antigen in the cardiac ganglia of dogs in paraffin sections. Tierarztl Prax, 19, 247-250.
Miranda NL, Robles CG, 1991. A comparative evaluation of a new immunozymatic test (RREID) with currently used diagnostic tests (DME and FAT) for dog rabies. Southeast Asian J Trop Med Public Health, 22, 46- 50.
Murphy FA, Bell JF, Bauer SP, Gardner JJ, Moore GJ, Harrison AR, Coe JE, 1980. Experimental chronic rabies in the cat. Lab Invest, 43, 231-241. Nietfeld JC, Rakich PM, Tyler DE, 1989. Bauer RW: Rabies-like inclusions in dogs. J Vet Diag Invest, 4, 333-338.
O’Brien DP, Axlund TW, 2005. Brain diseases. In: Textbook of Veterinary Internal Medicine, Diseases of the Dogs and Cat, Eds; Ettinger SJ, Feldman EC, Elsevier Saunders, St Louis, Missouri, USA, pp: 803-835.
Palmer DG, Ossent P, Suter MM, Ferrari E, 1985. Demonstration of rabies viral antigen in paraffin tissue sections: Comparison of the immunofluorescence technique with the unlabeled antibody enzyme method. Am J Vet Res, 46, 283-286.
Pedroso PMO, Pescador CA, Bandarra PM, Raymundo DL, Borba MR, Wouters F, Bezerra-Junior PS, Driemeier D, 2008. Standardization of immunohistochemistry technique for detection of rabies virus in formalin-fixed and paraffin-embedded tissue samples from central nervous system of cattle. Pesqui Vet Bras, 28, 627-632.
Pedroso PMO, Colodel EM, Pescador CA, Arrudo LP, Driemeier D, 2009. Clinical and pathological aspects in cattle affected by rabies with special reference to the rabies antigen mapping by immunohistochemistry. Pesqui Vet Bras, 29, 899-904.
Roseveare CW, Goolsby WD, Foppa IM, 2009. Potential and actual terrestrial rabies exposures in people and domestic animals, upstate south Carolina, 1994-2004: A people and domestic animals, upstate South Carolina, 1994-2004: A surveillance study. Potential and Actual Terrestrial, BMC Public Health, 9, 65.
Rudd RJ, Smith JS, Yager PA, Orciari LA, Trimarchi CV, 2005. A need for standardized rabies-virus diagnostic procedures: Effect of cover-glass mountant on the reliability of antigen detection by the fluorescent antibody test. Virus Res, 111, 83.
Rupprecht CE, Childs JE, 1996. Feline rabies. Feline Pract, 24, 15-19. Sinchaisri TA, Nagata T, Yoshikawa Y, Kai C, Yamanouchi K, 1992. Immunohistochemical and histopathological study of experimental rabies infection in mice. J Vet Med Sci, 54, 409-416.
Suja MS, Mahadevan A, Sundaram C, Mani J, Sagar BC, Hemachudha T, Wacharapluesadee S, Madhusudana SN, Shankar SK, 2004. Rabies encephalitis following fox bite, histological and immuno-histochemical evaluation of lesions caused by virus. Clin Neuropathol, 23, 271-276.
Sumedha 2010. Antimortem and post-pmortem detection of Rabies virus antigen in natural cases of rabies in animals-an immunopathological study. M.V.Sc. thesis, Guru Angad Dev Veterinary and animal Sciences University, Ludhiana, India.
Woldehiwet Z, 2005. Clinical laboratory advances in the detection of rabies virus. Clin Chim Acta, 351, 49-63.