Şap virüsü şuşlarına karşı poliklonal antikor üretimi

Antikorlar tanı laboratuvarlarında daima önemli bir yer tutmaktadır. Enzim işaretli bağışıklık testleri (ELISA), immunofloresan, antikor-mikroarray platformları, immunoblotlar, X-ışını kristalografisi gibi birçok tanı tekniği kullanımı antikorların varlığını gerektirir. Şap hastalığı serolojisinde antikorların kullanıldığı temel test ELISA’dır. ELISA testleri Şap virusu antijenini veya şap virusuna karşı organizmada oluşan antikorları tespit etmek için poliklonal veya monoklonal antikorlara ihtiyaç duyar. Bu amaçla, solid faz kompetetif ELISA (SPCE) ve likit faz bloking ELISA (LPBE) ve NSP ELISAlar kullanılır. Şap virusunun yapısal proteinlerine (SP) karşı oluşan antikorları tespit etmek için ise SPCE ve LPBE testleri kullanılır. Bu çalışma, ANep84 (genotip VII), O TUR07 (O Pan Asia II), ATUR11 (A Iran05) şap virusu suşlarına karşı (LPBE) testinde kullanılmak üzere poliklonal antikor üretmek amaçlandı. Bu amaçla, her bir FMDV serotipi için 4 kobay ve 6 tavşan kullanıldı. Antikorlar üretildikten sonra, dama tahtası (checkerboard ELISA titration) testi ile üretimi yapılan poliklonal antikorlar için optimal dilusyonlar belirlendi. Şap virusu suşlarına karşı oluşabilecek çapraz reaksiyonlar ve background reaksiyonları kontrol edildi. Sonuç olarak, bu çalışmada şap virusu ANep84 (genotip VII) ve O Tur07 (O Pan Asia II), ATUR11 (AIran05) suşlarına karşı poliklonal antikorlar üretildi ve LPBE testi için standardizasyonları yapıldı.

Anahtar Kelimeler:

şap hastalığı, üretim, poliklonal antikorlar, ELISA

Production of polyclonal antibodies against Turkey FMDV strains

Antibodies(Abs) have been always a major place in diagnostic laboratories. Many diagnostic techniques like Enzyme-Linked Immuno Sorbent Assays (ELISA), immunofluorescence, Ab-microarray platforms, immunoblots, X-ray crystallography require the Abs. ELISA is the main test that used Abs for Foot and Mouth Disease (FMDV) serology. It needs polyclonal or monoclonal Abs to detect FMDV antigen or Abs. For this purpose, solid-phase competitive ELISA (SPCE) or liquid phase blocking ELISA (LPBE) and non-structural protein (NSP) ELISA are used. SPCE and LPBE have mainly used FMDV structural protein (SP)-Ab detection. In this study, it was aimed to produce a polyclonal Ab against FMDV ANep84 (Genotype VII) and OTUR07 (OPanAsia II), ATUR11 (A Iran05) strains for LPBE, FMDV SP-antibody detection. For this purpose, 4 guinea pigs and 6 rabbits were used for each serotype of FMDV. After producing Abs, checkerboard ELISA titration was performed to determine the optimal test dilution of Abs. Backgrounds, cross-reactions against three strains of FMDV were also checked. In conclusion, polyclonal Abs were produced against FMDV ANep84 (Genotype VII) and O Tur07 (O Pan Asia II), ATUR11 (A Iran05) strains, and standardized for LPBE test.

Keywords:

FMD, Production, polyclonal antibodies, ELISA,

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