不同亚型禽流感病毒红细胞凝集特性分析

目的:了解不同亚型禽流感病毒与不同动物来源红细胞的凝集特性，为流感环境样本监测工作选择更适宜的检测用红细胞。方法:选取2009-2016年我国禽流感环境监测中分离到的不同亚型的禽流感毒株，采用红细胞凝集试验，选用5种动物红细胞(鸡、火鸡、豚鼠、马和绵羊)进行检测；应用流式细胞仪检测不同动物红细胞表面唾液酸受体的表达及类...

Analysis of erythrocyte agglutination characteristics of different subtypes of avian influenza virus

Objective To understand the agglutination characteristics of different subtypes of avian influenza viruses,we selected erythrocytes from different sources to find suitable erythrocytes for influenza environmental sample detection.Methods Different subtypes of avian influenza viruses,which were isolated from environmental sample between 2009 and 2016 in China,were selected to do hemagglutination assay using 5 animal erythrocytes(chicken,turkey,guinea pig,horse,and sheep).Flow cytometry was used to detect expression level and type of sialic acid receptors of different erythrocytes,and the characteristics of the receptor binding domain(RBD)of the viral hemagglutinin protein were analyzed by amino acid sequence.Results In this study,a total of 28 strains of avian influenza virus including 14 subtypes were detected.The result showed that all viruses could agglutinate with turkey and guinea pig erythrocytes and the rest three erythrocytes were unable to produce agglutination with some viruses;among them,one H9N2 virus(A/environment/Anhui/43762/2015)did not agglutinate with chicken erythrocytes,one H1N1 virus(A/environment/Shandong/76972/2014)and two H9N2 viruses(A/environment/Chongqing/79449/2014 and A/environment/Anhui/43762/2015)did not agglutinate with horse erythrocytes,two viruses of H9N2(A/environment/Chongqing/79449/2014 and A/environment/Anhui/43762/2015)and two viruses of H13N8(A/environment/Qinghai Lake/166/2012 and A/environment/Qinghai Lake/13/2012)did not agglutinate with sheep erythrocytes.The result of flow cytometry showed that two sialic acid receptors,α-2,3 andα-2,6,were detected on the surface of erythrocytes of turkey,chicken and guinea pig,but the expression ratios of the two receptors were different.Only the expression ofα-2,3 sialic acid receptors was detected in horse and sheep erythrocytes.Sequence analysis suggested that amino acid substitution in key regions of viral hemagglutinin protein RBD may be an important factor affecting the binding properties of different erythrocytes.Conclusions Our result suggested that turkey and guinea pig erythrocytes are the most sensitive in the hemagglutination test.Receptor expression and type of erythrocytes from different sources can significantly affect the agglutination reaction of different subtypes of avian influenza virus,and the amino acid changes in key regions of RBD can also affect the result of agglutination reaction.