流感病毒受体结合特性鉴别方法的建立

Development of A Method for Identification of Receptor Binding Specificity of Influenza Virus

目的建立一种快速鉴别流感病毒受体结合特性的方法。方法应用流式细胞仪技术,通过地高辛标记的植物凝集素DIG-SNA和DIG-MAA,检测正常鸡红细胞、绵羊红细胞和经α-2,3唾液酸酶处理的鸡红细胞表面受体。分别应用两种受体类型不同的红细胞进行微量血凝试验,检测流感病毒的受体结合特性,并检测醛化红细胞对其受体结合特性的影响。结果流式细胞检测结果表明,鸡红细胞表面既有SAα2,6Gal受体,也有SAα2,3Gal受体,绵羊红细胞表面只有SAα2,3Gal受体,处理后的鸡红细胞表面只有SAα2,6Gal受体。经分别检测人禽流感病毒毒株,证实该方法能快速准确地鉴别流感病毒的受体结合特性,醛化红细胞未影响其受体结合特性。结论已成功建立了流感病毒受体结合特性的鉴别方法。

Objective To develop a method for rapid identification of receptor binding specificity of influenza virus. Methods The receptors on surfaces of normal chicken red blood cells （RBCs）, the chicken RBCs treated with α-2, 3 sialidase and sheep RBCs were tested by flow cytometry with phytoagglutinin labeled by digoxin, i.e. DIG-SNA and DIG-MAA. Determine the receptor binding specificity of influenza virus, as well as influence of hydroformylation of RBCs on receptor binding specificity, by hemagglutination（HA） test with RBCs with two types of receptors. Results Flow cytometry showed both SAα2, 6Gal and SAα2, 3Gal receptors on surface of chicken RBCs, SAα2, 3Gal receptor on surface of sheep RBCs, and SAα2, 6Gal receptor on surface of chicken RBCs treated with α-2, 3 sialidase. Both the determination results of human and avian influenza virus strains proved that the developed method may be used for rapid and accurate identification of receptor binding specificity of influenza virus. The hydroformylation of RBCs showed no significant influence on receptor binding specificity. Conclusion A method for identification of receptor binding specificity of influenza virus was successfully developed.