Mükerrem GÜLEY, Mehmet TUNUS, Mehmet TOKER, Mediha ŞENTÜRK

Floresan Antikor Tekniği ile Kuduz Teşhisi

Bu mesaide, Floresan Antikor tekniği ile çalışmak için lüzumlu olan maddelerin istihsalleri ve kuduzdan şüpheli hayvan ve insan beyinlerinin FA tekniği ile boyanma metodundan bahsedilmektedir. .. Üç merkepten istihsal edilen kuduz antiserumlan titre edilerek uygun titre verenlerden gamma globulin elde edilmiş, bu gamma globulin iki ayrı florescein izotiocyanate ile konjuge edilerek, normal beyaz farelerden hazırlanan beyin ve karaciğer doku pudraları ile absorbe edilmiştir. Absorbsiyonu müteakip titresi yapılmış ve sonraki boypmalarda bu titre kullanılmıştır. Muhtelif cins hayvana, 13 adedi insana ait olmak üzere 809 adet marazi materyal floresan antikor tekniği, Sellers boyası ve fare iriokülasyonu ile muayeneye tabi tutulmuştur. Neticeler virus izoİasyonu esas alınmak suretiyle değerlendirilmiştir. 809 marazi materyalin 450 adedi, virus İzolasyonu, 443 adedi FA tekniği ile 374 adedi ise Sellers .metodu ile müsbet sonuç vermiştir. Burada Negri cisimlerinin tesbiti fare inokülasyonu ile %' 83.1 mutabakat gösterdği hadle, FA tekniği %98.4 mutabakat göstermiştir. Gliserin veya herhangi bir antiseptik içinde gönderilen marazi materyalin FA tekniği ile boyanınasında aspesifik reaksiyonların çokluğu yanıltıcı sonuçlar vermektedir. Buna mukabil tefessühten dolayı diğer metodlarla muayeneye elverişli olmıyan marazi materyal FA tekniği ile boyanabilmekte ve mükemmel neticeler alınmaktadır. Mevcut araştırmalar ve bizim çalışmamızın sonucu olarak diyebiliriz ki FA tekniği yetişmiş eleman, uygun ışık kaynağı ve iyi hazırlanmış bir konjugat sayesinde kuduz teşhis laboratuvarları için ideal bir metoddur.

Anahtar Kelimeler:

Floresan Antikor Tekniği, Kuduz, Kuduz Teşhisi

Rabies Diagnosis by Flurescent Antibody Technic

The diffuculty of the routine use of the FRA technique is the unavailability of commercially produced reagent. For this reason we started in producing conjugate, using three different löts of antisera and two different lots of fluorescein izothiocyanate, then preparation of tissue powders, sorption of serum globulins and titration of conjugate, also the procedure for staining rabies virus antigens with fluorescent labeled antibody in the brain of naturally infected animals is described. The usefulness of FRA test for routine diagnosis of rabies laboratory, particularly in the case of Negri - negative animals is emphasized. The brain tissues of 809 specimen, including dogs, cats cows, buffalos, donkies, horses, sheep, goats, wolves, squirells; mice, rabbits, hens, foxes, weasel, pig, hamster and 13 men were processed with fluorescent antibody. In addition to this test Sellers staining was carried out for Negri bodies and virus isolation studies were made on the brains. Results were evaluated on the base of virus izolation test. Totally 450 speciinen were found positives by virus isolation from the brains and 443 positives detected by FRA test, giving a 98:4 percent correlation. In contrast with this, we found 374 out of 450 positives. or 83.1 percent Negri bodies were demonstrable. If the brain tissues are preserved in glycerol- saline or in any other ·antiseptic, give aspecific reaction with FRA examination. On the other hand, this test gives satisfying result, even with decomposed specimens. Although same authors reports FRA test is 100 percent in accordaiıe with virus isolation, We could not get the same result. Because at the beginning we had not enough experience. But as soon as our experience improwed,. we got. nearly the· same· result with virus isolation. According to our study and recent literatures, FRA test is an ideal method for rabies diagnosis. A good result depends on a spesific light, well prepared reagets and experience4 persons.

PDF

___

1- Atanas:u, P : Quantitative Ass ay and. Poi:.ency. Test of Antira bi es Se. ruın. Laboratory Teckn.icjues in Rabies.; W. H. 6 .ivfonograph ~eries, No. 23, S. 167, Genova (1966).
2 - ''Batty, I., 'Walker-, P . D.: 'The · Use the Fluorescent Labelled Antibody Technique for the Detection and Differentiation of Bacteria.l Species., Internatıonial Sym·p. of Iminunol. Methods of Bi oL Standardization, Royaumont 1965; . SymP- : Series Imnobiol. Standard., VoL 4, S. 73-96, (Karger, Basel/New~York . 1967).
3 - Beauregard, M., Boulonger, P. and \-Vebster;· V. A. : The' Use of Fluores: .,_cent Antibody Staining ın , the Diagnosis of Ra~ıes., Ca:ıad. Jour. Comp. ı.'fed., 29, 1~;1-147 (1965)
4 - Carski, M. D., Wilsnack, R. E. Sikes, R; K. Pathogenesis of Rabies in . Wildltfe .. II. · Finarescent Antibody ,Studies., Amer . . Jour. Vet. Res., 23, 1048-1051, (1962)
5 - Cheny,. W. B.,. et Al, . : ~'luorescent Antibody· Technlques, in .. the Dlagjo nosis of Communicable Diseases., Bureau of State Servlces Communicable Disease Center .. Atlanta, Georgia, (1960).
6 - Coons, A. H., et Al. : The Demoristration of Pneumococcal Antıgen in 'l'issues by the Use of Fluorescent Antibody., J. Immuno!. 45, 159-170, {1942).
7 - (;oons, A. H., anıt Kaplan, l\1. H. : Localizatlon of Antıgen in Tissue Cell3. IL Improvements in a Methode for the Detectıon of Antigen by Means of Fluorescent Antibody., Jour. Exp. Med., 91, 1-13, (1950) .
8 - Cooııs, A. H., et Al. : Localization of Antigen In Tlssue Cells. IV. Antig-!n of Rickettsiae and Mumps Virus., Jour. Exp. Med., 91, 31-37, (1950).
9 - Dean, D. J. : The Fluorescent Antibody Test., Laboratory Techniques in Rabies, World Health Organization, Monograph Serie!.l No. 23 Geneva, (1966).
10 - Etc.hebarne, M., Bernal, P. G., :Leyton, G. R. : Purificatıon of Rabies Antıbodies in Hors Serum and Diagnostic Importance of the Fluorescent
11- Goldwasser, R. A~, and Kissling, R. E.: Fluorescent Antibody- Staining of Street and Fixed Rabies Virus Antigens., Proc. Soc. Exp. Biol. and Med., 98, 219-223, (1958).
12- Goldwasser, R; A., Kissling, · R. E., . Carski, T. R. : Fluorescent antibody Staining of Rabies Antigens in the Salivary Glands of Rabid Animals., Reprinted From Bul!. Wld Hlth Org.", 20, 579-588 (1959).
13 - Güley, M. : Kuduz Laboratuvarının GÜnlük Çalışmaları., Etlik · Vet. Bakt. Enst. Der., ı, .38-42, (ı963).
14 - Gtilmezoğlu, E. : Çocuk İshallerinde Enteropathogenic E . Coli İdantifikasyonunda Floresan Antikor Tekniğinin Kullanılması., Çocuk Sağlığı ve Hastalıkları Dergisi, Cilt, 6 (4), 206, (ı963).
15 - Gtilmezoğlu, E. : Floresaıi Antıkor Tekniği., Türk Hijiyen ve Tecrübi Biöloji Dergisi, Cnt XXIV, S. 2, ı8ı-ı97, (ı964).
16-Johnson, H. N., Edited by Rivers, T. :M. : Viral and Rickettsial Infecttions of Man., S. 267-299, Lippincott Co., Philadelphia, (ı952).
17 - Kaplan, M. H., Coons, A. H., and Dean, H. \V.: Localization of Antigen in Tissue Cells. III. Cellular Distribution of Pneumococcal Polysaccarides Types Il anel III in the Mousa., Jour. Exp. Med ~ . 9ı, ı5-29, "(1950) .
18 - Lepine, P., Atanasıu, P~ : _Production of .Therapeutic Antisabies Serum., labora:tor,:y; tehniques in Rabies· World Health Organization, Monograph Series No. 23," Geneva, (ı966).
19 - Mars~all, . J. D., Eveıand , W. C.,. and Smith, C. W. : Superiority of FIIC (Riggs) for Fluorescent ~- Antibody Technic With a Modi!ica tion of !ts Application., Proc. Soc. Exp. Biol. and Med., .98, 898-900, (ı958).
20-- Mc Queen, J . L. : Rabies Diagnosis Special Applicatıon of Fluorescent Antibody Tecniques., Reprinted from Proceed!ng United States Livestock Sa,nitary Assqciati011., ( ı9Ş9) .
21- Mc QtİeeN,' .J. L. Le\ris~ A: L., and Schneider, N. J. : Rabies Diagnosis by Fluorescent Antibody. I. !ts Evaluation a Public Health Laboratory., Amer. Jo.ur. P~b~. - Health,- 50, 1743-1752, {1960). . .
22 ~ Rig~, J. L., et Al:: Izothiocyanate Compounds as ·Fluorescent Labeling Agents for Immune Serum., Arner. Jour. Path., 34, ıo8ı, (1958) .
23 -Sikes, R~ K.·: Avaluiı.tion of Anti•Rabies Conjugates .. Reprint, · Aslı bulunamadı;
24 - Wilsna.ck, R. E., : The Fluorescent Antıbody Diagnosis of Rabies, Reprinted . from Jour. Amer .. Vet. Med. Assoc., Vol. ı37, No. 5, ı, (ı960). Antibody Technlque., Jour. Immuno!., 84, 6-10, (1960).

Etlik Veteriner Mikrobiyoloji Dergisi-Cover

ISSN: 1016-3573
Yayın Aralığı: Yılda 2 Sayı
Başlangıç: 1960
Yayıncı: Veteriner Kontrol Merkez Araştırma Enstitüsü Müdürlüğü

Arşiv