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Tavuk Hastalıklarının Teşhisinde Kullanılan İmmunofloresans Tekniği İçin Konjugeyt Hazırlanması

Year 1997, Volume: 9 Issue: 1, 139 - 148, 01.12.1997

Abstract

İmmunfloresan veya floresan antikor metodu (IF) antijenleri identifiye etmek
antijenlerin yerini tayin etmek antikor tayin etmek ve mikroskop altına görünür
hale getirmek için işaretlenmiş antikor veya antijenlere uygulanan bir tekniktir.
işaretli antikorların kendi antijenine bağlanması, spestifik bir antijen-antikor
reaksiyonudur. IF tekniği, bir immunolojik reaksiyonun hassasiyet özelliğini
mikroskop doğruluğu ile birleştirir ( 1 5). Antijen-antikor bağlanmaları
immunfloresan tekniğinde değişik metodlarla yapılabilir. Direkt ve indirekt
teknikleri özellikle ve hassasiyet ve yönünden karşılaştırıldıklarında, floresan
madde ile işaretli antiserumla yapılan direkt teknikleri, anti inmmnoglobulin
kullanarak yapılan tekniklerden 2 kat daha az hassas olduğu ortaya konulmuştur
(4). IF metodu ilk defa Coons ve arkadaşları tarafından 1942 yılında bulunmuş
( 13) ve daha sonra geliştirilerek geçtiğimiz yıllarda ve günümüzde yaygın olarak
mikrobiyoloji, viroloji, immunoloji ve biyoloji alanlarında kullanılmaktadır.
Şimdiye kadar ülkemizde özellikle tavuk hastalıklarının teşhisinde kullanımı,
tekniğin ikinci safhasında kullanılan konjugeytin bulunmaması ve üretilmemesi
yüzünden sınırlı kalmıştır. Keza bu tür reagentleri üreten şirketlerde az sayıdadır.
Bu çalışmada tavuk hastalıklarının indirekt floresan antikor tekniği ile teşhis
edilmesi amacıyla konjugeyt üretimi yapılmıştır. Ülkemizde tavuk hastalıklarının
teşhisi ve araştım1a çalışmalarında dışarıdan oldukça zor temin edilerek kullanılan
konjugeytin üretilmesiyle bu test daha yaygın bir şekilde kullanılacaktır.
Bu araştım1ada tavşan antitavuk iımnunoglobulinin floresan izotiyosiyanat
ile işaretlenme metodu çalışılmıştır. Çalışma ayrıca; tavuk lg G elde edilmesini,
anti-antikor üretecek tavşanların immunizasyonunu tavşan anti-tavuk IgG elde
edilmesini, FITC ile işaretlenmesi tekniklerini kapsamaktadır.
Tavuk Hastalıklannda İmınunofloresans-BAYRAKTAR-BAYRAKTAR-DAGISTAN-ALAY

References

  • ARDA, M. (1971): Hastalık etkenlerinin titrasyon ve nötralizasyon testlerinde uygulanan laboratuvar metodları . A . Ü . Vet.Fak.Yayın no:273 .
  • BRADFORD, M. M. ( 1976) : A.rapid and sensitive method for the quantitation of microgram quantities of protein Utilizing the principle of Protein Dye binding. Analytical Biochemistıy 72 :248-254.
  • GOLDMAN, M. (1968): Fluorcscent Antibody methds. Bionctics Research, USA. Academic Press . 87-148.
  • HAAIJMAN, J. J., SLINGERLAD, T. ( 1978) : Equipmend and Preparative procedures in lmmuno-fluorescense microscopy: Quantitative studies . lmmunofluorcscense and rclated staining technigues. Elsevier/ NortHolland Biomcdical Press . 1 1-28 .
  • HARLOW, E., LANE, D. ( 1988) : Labeline antibodies with fluorochromes antibodies. A laboratory Manual. 2nd Edition Cold Spring Harborn laboratory 353-358.
  • HEBERT, G. A., PATRICIA, L. P. , PITTMAN, B.( 1973) : Dctermination of the optimal Ammonium Sulphate Concentration for the Fractionation of Rabbit, Sheep, Horse and Coat Antisera, Applied Microbiology. 25 : 26-36 .
  • HOLBOROW, E.J., JOHNSON, G.D. ( 1 976) : Application of Imımınofluoresccnsc . Immunofluorescense ll. Ed. Elsevier Press. Chov : 18.
  • JEFFREY, LS. ( 1 982) : Fluorescencemicroscopy. International Laboratory Vol: 12. No. 8:46-52.
  • JOHNSON, G.D., DOLLHOPF, F.L. ( 1968) : Filter systems for flurescent Antibody techniques. lmmınology 15 : 619-621 .
  • NAIRN, R.C. (1969): Fluorochromes and their conjugation with proteins. Fluorescent protein Tracing. E.S.Livingstone LTD.3.rd Edition
  • NOWOTNY, A. 1969: Bacic Exercises in lmmunochemistry. Tample University. scool of Medicine, Philadelpia. 2nd Edition .
  • PHYLAXIA Veterinary Research Institute, Hungary. ( 1987) : Viral products methods booklet 92-1 18.
  • SKVARIL, F., FRAGNER, J. ( 1965): Experiences with the prcparation of Animal Antiglobulins labcllcd with tlurcsccin isothiocyanate (FITC).Journay of Hygicne, Epidcıniology, M icrobiologoy and lımnunology 9:5 15-523.
  • TOMLINSON, A.H. ( 1 967) : Quartz-iodine illumination. Imınnology 13 : 323-324.
  • TRAUTWEIN, G. (1986): Quatine oflınımınofluoresccnse. Handbook of laboratory techniques. Institute of Pathology Veterinary school Hannaver FR.G.28-57 .
  • VİLLEGAS, P. ( 1987): Application of lmmunofluorescence. Avin Vırus Disease, Laboratory Manual. College of veterinary Medicine, Universitiy of Georgia. 55-70.
  • VUNAKIS, H. V., LANGüN E, J.J. ( 1980) : Immunochemical Techniques. Methods in Enzymology. Academic Press. 49-104.
  • WEYBRİDGE Central Yeterinary Laboratory, U. K. (1988) : Diagnostic products control seetion method shcets. 18-35 .
  • WlLAMELIN, J.P., LENOIR, G. ( 1978): Prcparation and conjugation to fluorescein isothio-cyanatc of immunoglobulins. Mamıal of tcclmiques in immunology· and virology for canccr research workcrs post graduate course booklet. lntcr. Ageney for Res. on cancer, Lyon-Francc, 239-254.
  • WOLLAGE, J.C.,DUGUID, J.P., FRASER,A.G., MARMION, B.P. (1989) : Labelled Antibodies, Practical Medical Microbiology. 13. th edition. Churchill livingstone Press, London, Vol :2 197-200.

The preparation of conjugate for the immunofluoresans techniques of the diagnosis of poultry diseases.

Year 1997, Volume: 9 Issue: 1, 139 - 148, 01.12.1997

Abstract

The method of immunofluorescence or fluorescent antibody is an
inununological teclınique which applies antibodies labelled with a fluorcscent
dye in order to visualize, identify or locate antigens. The binding of labelled
antibody its antigen is a specific antigen-antibody rection. The IF technique
combines the sensitivity and specifity of an inununological reaction with the
precision of microscopy ( 1 5). The IF method which was elaborated by Coons ct
al (13) in 1942 and was improved later, has come to be used very widely in
recent years especially in some of the western countries It is used in microbiology,
immunology and general biology. The binding of antibadies to antigens may be
visualized in immunofluoresccnce.
In this study, we have produced conjugate for indirect FAT methods for
poultry diseases. With the aim of making the conjugate for diagnosis of poultry
diseases and research activities in Turkey will be available more easily.
A method of FITC labelled anti IgG is described in here. The technique
consists in the preperation of chicken lgG, immunization of producer animals,
the preparation of rabbit antichicken IgG and the conjugation of rabbit antichicken
IgG with FITC and fınal treatment of the preparation. The quality of the fınal
product is regularly tested in standart methods.

References

  • ARDA, M. (1971): Hastalık etkenlerinin titrasyon ve nötralizasyon testlerinde uygulanan laboratuvar metodları . A . Ü . Vet.Fak.Yayın no:273 .
  • BRADFORD, M. M. ( 1976) : A.rapid and sensitive method for the quantitation of microgram quantities of protein Utilizing the principle of Protein Dye binding. Analytical Biochemistıy 72 :248-254.
  • GOLDMAN, M. (1968): Fluorcscent Antibody methds. Bionctics Research, USA. Academic Press . 87-148.
  • HAAIJMAN, J. J., SLINGERLAD, T. ( 1978) : Equipmend and Preparative procedures in lmmuno-fluorescense microscopy: Quantitative studies . lmmunofluorcscense and rclated staining technigues. Elsevier/ NortHolland Biomcdical Press . 1 1-28 .
  • HARLOW, E., LANE, D. ( 1988) : Labeline antibodies with fluorochromes antibodies. A laboratory Manual. 2nd Edition Cold Spring Harborn laboratory 353-358.
  • HEBERT, G. A., PATRICIA, L. P. , PITTMAN, B.( 1973) : Dctermination of the optimal Ammonium Sulphate Concentration for the Fractionation of Rabbit, Sheep, Horse and Coat Antisera, Applied Microbiology. 25 : 26-36 .
  • HOLBOROW, E.J., JOHNSON, G.D. ( 1 976) : Application of Imımınofluoresccnsc . Immunofluorescense ll. Ed. Elsevier Press. Chov : 18.
  • JEFFREY, LS. ( 1 982) : Fluorescencemicroscopy. International Laboratory Vol: 12. No. 8:46-52.
  • JOHNSON, G.D., DOLLHOPF, F.L. ( 1968) : Filter systems for flurescent Antibody techniques. lmmınology 15 : 619-621 .
  • NAIRN, R.C. (1969): Fluorochromes and their conjugation with proteins. Fluorescent protein Tracing. E.S.Livingstone LTD.3.rd Edition
  • NOWOTNY, A. 1969: Bacic Exercises in lmmunochemistry. Tample University. scool of Medicine, Philadelpia. 2nd Edition .
  • PHYLAXIA Veterinary Research Institute, Hungary. ( 1987) : Viral products methods booklet 92-1 18.
  • SKVARIL, F., FRAGNER, J. ( 1965): Experiences with the prcparation of Animal Antiglobulins labcllcd with tlurcsccin isothiocyanate (FITC).Journay of Hygicne, Epidcıniology, M icrobiologoy and lımnunology 9:5 15-523.
  • TOMLINSON, A.H. ( 1 967) : Quartz-iodine illumination. Imınnology 13 : 323-324.
  • TRAUTWEIN, G. (1986): Quatine oflınımınofluoresccnse. Handbook of laboratory techniques. Institute of Pathology Veterinary school Hannaver FR.G.28-57 .
  • VİLLEGAS, P. ( 1987): Application of lmmunofluorescence. Avin Vırus Disease, Laboratory Manual. College of veterinary Medicine, Universitiy of Georgia. 55-70.
  • VUNAKIS, H. V., LANGüN E, J.J. ( 1980) : Immunochemical Techniques. Methods in Enzymology. Academic Press. 49-104.
  • WEYBRİDGE Central Yeterinary Laboratory, U. K. (1988) : Diagnostic products control seetion method shcets. 18-35 .
  • WlLAMELIN, J.P., LENOIR, G. ( 1978): Prcparation and conjugation to fluorescein isothio-cyanatc of immunoglobulins. Mamıal of tcclmiques in immunology· and virology for canccr research workcrs post graduate course booklet. lntcr. Ageney for Res. on cancer, Lyon-Francc, 239-254.
  • WOLLAGE, J.C.,DUGUID, J.P., FRASER,A.G., MARMION, B.P. (1989) : Labelled Antibodies, Practical Medical Microbiology. 13. th edition. Churchill livingstone Press, London, Vol :2 197-200.
There are 20 citations in total.

Details

Primary Language Turkish
Subjects Veterinary Surgery
Journal Section Original Article
Authors

Ragıp Bayraktar This is me

Lale Bayraktar This is me

Mehmet Dagıstan This is me

Osman D. Alay This is me

Publication Date December 1, 1997
Submission Date December 1, 1997
Published in Issue Year 1997 Volume: 9 Issue: 1

Cite

APA Bayraktar, R., Bayraktar, L., Dagıstan, M., Alay, O. D. (1997). Tavuk Hastalıklarının Teşhisinde Kullanılan İmmunofloresans Tekniği İçin Konjugeyt Hazırlanması. Etlik Veteriner Mikrobiyoloji Dergisi, 9(1), 139-148.

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