将鸡伤寒沙门氏菌LPS抗原包被于渗滤盒检测区(T区),针对沙门氏菌O9抗原单克隆抗体3-47-0包被在质控区(C区),用胶体金标记鸡伤寒沙门氏菌LPS抗原,建立了一种抗原夹心法快速检测鸡白痢、鸡伤寒沙门氏菌抗体的斑点免疫金渗滤法(DIGFA)。DIGFA比玻板凝集试验(PAT)灵敏度高,人工感染鸡试验表明在感染后第7d即可从32只鸡中的7只检测到抗体,在时间上早于PAT。722份现场血清样品的检疫结果表明,DIGFA的阳性检出率稍高于PAT,阳性样品抗体几何平均滴度DIGFA大于PAT法(P〈0.05)。DIGFA的结果得到细菌分离试验的验证。这些结果表明DIGFA快速、灵敏、准确,为鸡伤寒和鸡白痢的检疫提供了一个新的有效手段。
Dot immuno-gold filtration assay (DIGFA) to detect antibodies against Salmonella pullorum/gallinarum was established by using lipopolysaccharide (LPS) from Salmonella gallinarum reference strain as coating and labeling antigen. DIGFA was more sensitive than two commercially available plate agglutination test (PAT) reagents, as shown by positive sample dilution experiment and artificial infection experiment. 722 samples were detected by DIGFA and two PAT reagents, the results showed DIGFA was more sensitive, which was further demonstrated by the positive serum antibody arithmetical average titer. The result of DIGFA was confirmed by bacterial isolation and identification. In summary, DIGFA is rapid, sensitive, specific, which offers a new method to screen fowl pullorosis and typhoid disease in chickens.