Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels

Aybay, Cemalettin; Atak Yücel, Ayşegül

Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels

Cemalettin AYBAY, (Gazi Üniversitesi, Tıp Fakültesi, İmmünoloji Anabilim Dalı, Ankara, Türkiye)

AYŞEGÜL ATAK YÜCEL (Girilmedi)

Gazi Medical Journal

1 0

Yıl: 2002 Cilt: 13 Sayı: 4 Sayfa Aralığı: 177 - 186 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels

Öz:

Amaç: Tüm dünyada ve Türkiye'de uygulanan rutin aşılama programları nedeni ile difterinin çok nadiren görülen bir hastalık olduğu düşünülüyordu. Ancak, eski Sovyetler Birliği'nden ayrılan Yeni Bağımsız Devletler'de görülen epidemi toplumsal bağışıklığın yeterli düzeyde olmayabileceği ve bu nedenle hastalığın yayılabileceği kaygısını uyandırdı. Hastalığı ortadan kaldırmaya yönelik aşılama programlarını başlatabilmek için ilk önce tüm yaş gruplarında difteri toksinine (DT) karşı serum antikor seviyeleri standardize edilmiş bir metod ile ölçülmelidir. Ancak ticari anti-DT ELISA kitlerinin maliyeti çok yüksektir. Bu çalışmada hem tam hem de tarama amaçlı kullanılabilecek yüksek duyarlılıkta ve özgüllükte, ancak düşük maliyetli bir anti-DT ELISA sisteminin yapılması amaçlanmıştır. Yöntem: Katı faz antijen olarak kullanılan difteri toksini ile kaplanırken konjugat olarak da enzimle işaretli DT kullanıldı. Serum ya da diğer örneklerdeki anti-DT antikorlar katı fazdaki ve konjıtgattaki DT'ye iki adet Fab uçları ile bağlandığından, oluşturulan sistem farklı türlere ait serum örneklerindeki özgül antikorları da ölçebilmektedir. Laboratuvarda hazırlanan bu ELISA sistemi ayrıca ticari bir anti-DT ELISA kiti ile de karşılaştırıldı. Bulgular: Ticari anti-DT ELISA kiti 0.1 IU/ml'nin altındaki antikor seviyelerini ölçemediği halde, geliştirilen sistemin 0.01 IU/ml altındaki antikor seviyelerini ölçebildiği ve dinamik ölçüm aralığının 0.01-1 IU/ml arasında olduğu saptandı. Sonuç: Hazırlanan anti-DT ELISA çok duyarlı, güvenilir ve düşük maliyette bir sistem olup, difteriye karşı bağışıklık durumunun değerlendirilmesinde kullanılabilecektir. Ayrıca, humoral immün yanıtla ilgili çeşitli in vivo ve in vitro araştırma çalışmalarında da kullanılabilecektir.

Anahtar Kelime: Duyarlılık ve özgüllük Enzim bağlı immünosorbent tayini Difteri antitoksini Antikor formasyonu Difteri toksini Difteri

Konular: Genel ve Dahili Tıp

Difteri anti-toksin seviyelerinin ölçülmesi için tanı ve tarama amaçlı bir elisa sistemi geliştirilmesi

Öz:

Purpose: It had been thought that diphtheria was a very rare disease because of routine immunization programs conducted worldwide. However, the epidemic seen in the newly independent states of the former Soviet Union prompted worries that public immunity levels might not be sufficient and therefore the disease might spread. In order to conduct vaccination programs to eliminate the disease, first of all, the serum antibody levels of all age groups to diphtheria toxin (DT) should be measured with a standardized method. However, the cost of commercial anti-DT ELISA kits is very high. In this study we aimed to develop an anti-DT ELISA system that could be used for both diagnosis and screening, with high specificity and sensitivity but low cost. Methods: Solid phase was coated with diphtheria toxin while enzyme-labeled DT was used as conjugate. Since the anti-DT antibodies in serum or other samples to be measured will bind DT in the solid phase and conjugate with its two Fab ends, the system can be used for sera from other species as well. This in-house ELISA was also compared with a commercially available anti-DT ELISA kit. Results: Although the commercial anti-DT ELISA kit had restrictions for measuring antibody levels below 0.1 IUf ml, the in-house ELISA system managed to measure anti-DT antibody levels below O. Ol IU/ml with a dynamic analytical measuring range between O. OJ and 1IU/ml. Conclusion: The in-house anti-DT ELISA system is a highly sensitive, reliable and low-cost test technique that can be used in the diagnosis and screening of immunity to diphtheria. In addition, it can be used in many different in vivo and in vitro investigations of humoral immune responses.

Anahtar Kelime: Diphtheria Toxin Diphtheria Sensitivity and Specificity Enzyme-Linked Immunosorbent Assay Diphtheria Antitoxin Antibody Formation

Konular: Genel ve Dahili Tıp

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

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APA	Aybay C, Atak Yücel A (2002). Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels. , 177 - 186.
Chicago	Aybay Cemalettin,Atak Yücel Ayşegül Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels. (2002): 177 - 186.
MLA	Aybay Cemalettin,Atak Yücel Ayşegül Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels. , 2002, ss.177 - 186.
AMA	Aybay C,Atak Yücel A Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels. . 2002; 177 - 186.
Vancouver	Aybay C,Atak Yücel A Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels. . 2002; 177 - 186.
IEEE	Aybay C,Atak Yücel A "Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels." , ss.177 - 186, 2002.
ISNAD	Aybay, Cemalettin - Atak Yücel, Ayşegül. "Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels". (2002), 177-186.

APA	Aybay C, Atak Yücel A (2002). Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels. Gazi Medical Journal, 13(4), 177 - 186.
Chicago	Aybay Cemalettin,Atak Yücel Ayşegül Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels. Gazi Medical Journal 13, no.4 (2002): 177 - 186.
MLA	Aybay Cemalettin,Atak Yücel Ayşegül Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels. Gazi Medical Journal, vol.13, no.4, 2002, ss.177 - 186.
AMA	Aybay C,Atak Yücel A Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels. Gazi Medical Journal. 2002; 13(4): 177 - 186.
Vancouver	Aybay C,Atak Yücel A Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels. Gazi Medical Journal. 2002; 13(4): 177 - 186.
IEEE	Aybay C,Atak Yücel A "Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels." Gazi Medical Journal, 13, ss.177 - 186, 2002.
ISNAD	Aybay, Cemalettin - Atak Yücel, Ayşegül. "Development of a diagnostic and screening elisa system for measuring diphtheria anti-toxin levels". Gazi Medical Journal 13/4 (2002), 177-186.