Jianhe Gaiping Zhiqiang Shuguang Lei Shouping Likun Huihui Xiaojing HU ZHANG SHEN LI WANG ZHANG CHENG ZHANG XIA

Diagnostik Antijen Olarak Rekombinant Sao-M Protein Kullanılarak Streptococcus suis'in Tanısı Amacıyla PPA-ELİSA Geliştirilmesi

Streptococcus suis, insanlarda enfeksiyona neden olan salgınlardan sorumlu önemli bir zoonotik ajandır. S. suis'in doğru ve hızlı bir şekilde tespit edilmesi enfeksiyona bağlı salgınları kontrol edebilir ve hastalığa bağlı etkileri azaltabilir. Bu çalışmada, Sao-M, S suis enfeksiyonunun teşhisi amacıyla peroksidaz enzim-bağlı stafilokokal protein A immün testi (PPA-ELİSA) için kullanılmıştır. Satranç tahtası titrasyon testinin sonuçları, kaplama antijeninin optimal konsantrasyonu ve serum sulandırılmasını sırasıyla 8 µg/mL ve 1:80 olarak göstermiştir. Pozitif cevap için eşik değeri OD450>=0.351 olarak belirlendi. Testin spesifitesi rSao-M'nin diğer 6 patojen türe karşı kullanılan antiserum ile çapraz reaksiyon vermediğini gösterdi. Tekrar testlerinde yığın içi ve yığınlar arası varyasyon katsayısı %9.5'ten az idi. Geleneksel ELİSA kiti ve kurulan GDH tabanlı ELİSA'nın karşılaştırmalı analizinde, mevcut PPA-ELİSA testinin GDH tabanlı ELİSA testinden daha yüksek spesifite ve sensitiviteye sahip olduğunu gösterdi. 500 domuz serumu örneğinde toplam seroprevalansın %6.6 olduğu ve bunun yöntemin S. suis enfeksiyonunun saptanmasında uygunluğunu gösterdi. Sonuçlar, PPA-ELİSA'nın hızlı, duyarlı ve spesifik bir tanı yöntemi olduğunu ve S. suis enfeksiyonunun serolojik tanısı için büyük kapsamlı epidemiyolojik araştırmalarda yeni bir araç olarak kullanılabilirliğini göstermektedir.

Development of PPA-ELISA for Diagnosing Streptococcus suis Infection Using Recombinant Sao-M Protein As Diagnostic Antigen

Streptococcus suis, an important zoonotic agent, is responsible for outbreaks of human infections. The accurate and rapid detection of S. suis may help control the potential outbreak and ameliorate patient outcomes. In the present study, Sao-M was used to establish a horseradish peroxidase enzyme-linked staphylococcal protein A immunosorbent assay (PPA-ELISA) for the diagnosis of S. suis infection. Results of chessboard titration test showed that the optimal concentration of coating antigen and dilution of serum were 8 µg/ml and 1:80, respectively. The cut-off was confirmed as OD450>=0.351 for positive response. The specificity of test indicated that rSao-M had no cross-reaction with antisera against the other 6 species of pathogens. The variation coefficient of intra-batch and inter-batch in the repeating tests was less than 9.5%. Comparative analysis by using conventional ELISA kit and established GDH-based ELISA showed that the present PPA-ELISA has higher specificity and sensitivity than GDH-based ELISA. A total seroprevalence of 6.6% in 500 pig serum samples indicated the method's applicability to detect S. suis infection. Cumulatively, the results suggested that the PPA-ELISA is a rapid, sensitive and specific diagnostic method and could be used as a new tool for large-scale epidemiological surveys and serological diagnosis of S. suis infection.

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