酶标单抗阻断ELISA检测鸡白痢和鸡伤寒抗体

A Blocking ELISA for the Diagnosis of Pullorum Disease and Fowl Typhoid by Using Peroxidase-Labelled Monoclonal Antibody and Salmonella pullorum Lipopolysaccharide

以辣根过氧化物酶标记的沙门氏菌O9单抗3-47-0与包被的鸡白痢沙门氏菌脂多糖抗原建立了一种抗体阻断EL ISA以检测鸡白痢和鸡伤寒沙门氏菌感染。该方法基于待检血清样品抑制酶标单抗与脂多糖的结合,以检测鸡白痢和鸡伤寒特异性抗体。对72份已知鸡白痢阳性血清(抑制率为(98.5±4.0)%)、54份已知鸡白痢阴性血清(抑制率为(15.3±8.0)%)、42份SPF鸡血清(抑制率为(0.8±7.2)%)检测的结果表明,该方法具有很强的区分能力。在人工感染试验中,从第2周开始,该方法能从全部鸡只中检测到特异性抗体,比全血玻板凝集试验(PAT)早1周时间。对344份种鸡血清的检测结果表明,单抗阻断EL ISA比PAT特异性好、敏感性高。另外,该方法也能检测出与鸡白痢和鸡伤寒沙门氏菌有共同抗原的沙门氏菌感染产生的抗体。

A blocking ELISA was developed for diagnosing pullorum disease and fowl typhoid. The test measured the inhibition of binding between a labeled,monoclonal antibody 3-47-0 and Salmonella pullorum lipopolysaccharide （LPS）. Good discrimination was observed between 72 proven pullorum disease sera with inhibition rate of（98. 5±4.0）% and 54 sera negative for pullorum disease with inhibition rate of （15.3±8.0）% and 42 sera of SPF chicken with inhibition rate of（10. 8±7.2）%. Chickens infected with S. pullorum were monitored serologically by this assay from one through eight weeks. From the second week after infection, 100% of the chickens were serologically positive by blocking ELISA,which was one week earlier than PAT. The high specificity and sensitivity of blocking ELISA was further verified by detection of 344 clinical sera with parallelly conducted tests（PAT）. Meanwhile,the assay potentially detects those systemic infections caused by Salmonella pullorum and Salmonella gallinarum that possess common O antigen.