ARZU FINDIK, ÖZLEM BÜYÜKTANIR YAŞ, Nevzat YURDUSEV

LPS and Flagellin-based models for serological screening and confirmation of Salmonella infections in chickens

Salmonella B ve D serogruplarına ait kanatlı salmonellosis’i halk sağlığı yönünden ve kanatlı endüstrisindeki ekonomik kayıplar nedeniyle büyük öneme sahiptir. Kanatlı salmonellozisinin etkin tanısı amacıyla, SE lipopolisakkarid (LPS) ve saflaştırılmış flagellin (Flg) tanı antijenlerine dayalı iki indirekt ELISA modeli geliştirilmesinde ticari Salmonella Enteritidis (SE) ELISA kiti ile değerlendirilmiş toplam 126 tavuk serumu kullanıldı. Optimizasyon sonrası, 154 serum indirekt LPS-ELISA ve 263 serum Flg-ELISA ile test edildi. Ticari LPS-ELISA kiti ile karşılaştırıldığında, indirekt LPS- ve Flg-ELISA’ların anti-Salmonella antikor tespit performansları yüksek ve benzer bulundu. Bununla birlikte, sınırlı sayıda serumun Flg- ve LPS-ELISA’da farklılık gösterdiği belirlendi. Bu uyumsuz serumların SE bakteri lizatı ile reaktif olduğu, buna karşın yalnızca LPS veya Flg antijenini tanıdıkları Western Blot analiziyle gösterildi. Bu çalışmadan, LPS- ve Flg-ELISA’nın eşzamanlı kullanımıyla kanatlılara özgü salmonellozisin flagellalı bakteri kökenli zoonotik Salmonella enfeksiyonlarından ayırıcı tanısının yapılabileceği ve Western blot modellerinin doğrulayıcı serolojik test olarak kullanılabileceği sonucuna varıldı.

Anahtar Kelimeler:

bağışıksal tanı, zoonoz, seroloji, salmonelloz, halk sağlığı, kümes hayvanları, tanı teknikleri, teşhis, antijenler, antikor, Salmonella enteritidis, ELISA

Tavuklarda Salmonella enfeksiyonlarının serolojik tanısı ve doğrulanmasına yönelik LPS ve flagelline dayalı modeller

Avian salmonellosis due to Salmonella B and D serogroups has a great importance because of the public health and the economical losses in poultry industry. For accurate diagnosis of avian salmonellosis, a total of 126 chicken sera evaluated with a commercial Salmonella Enteritidis (SE) ELISA kit have been used to develop two indirect ELISA models using lipopolysaccharide (LPS) and purified flagellin (Flg) of SE as diagnostic antigens. After optimization, 154 and 263 sera were tested with indirect LPS- and Flg-ELISA, respectively. The performance of both indirect LPS- and Flg-ELISA was found high and similar for detecting anti-Salmonella antibody in comparison with a commercial LPS-ELISA kit. Nevertheless, a limited number of chicken sera were found discordant between Flg- and LPS-ELISAs. Western-blot analyses of the discordant sera demonstrated that they were found reactive with bacterial lysate of SE but only recognized LPS or Flg antigen. It was concluded from this study that the simultaneous use of both LPS- and Flg-ELISAs would allow differential diagnosis of avian specific salmonellosis from zoonotic Salmonella infections due to flagellated bacteria and Western blot models can be used as confirmatory serological test.

Keywords:

immunodiagnosis, zoonoses, serology, salmonellosis, public health, poultry, diagnostic techniques, diagnosis, antigens, antibodies, Salmonella enteritidis, ELISA,

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