MHC-B单倍型BW/G系鸡群同种异型抗血清的制备及应用  

Preparation and Application of Allotype Antisera for MHC-B Complex in BW/G Lines Chicken

作　　者：杨柳[1,2] 高彩霞[2] 武永淑[2] 李海超[1,2] 杨超[2] 廉传江[2] 韩凌霞[2] 

机构地区：[1]东北农业大学,黑龙江哈尔滨150030 [2]中国农业科学院哈尔滨兽医研究所兽医生物技术国家重点实验室/实验动物中心,黑龙江哈尔滨150001

出　　处：《中国家禽》2013年第14期14-18,共5页China Poultry

基　　金：国家自然科学基金项目(31101800)

摘　　要：BW/G(n)系鸡群是主要组织相容性复合体核心区纯合的SPF鸡群,包括BW/G1、BW/G2、BW/G3、BW/G5、BW/G6和BW/G76个亚系,分别属于B13、B15、B2、B5、B21、B19单倍型。本研究以BW/G(n)系鸡的外周血红细胞作为免疫原,静脉接种其它不同单倍型鸡,制备了同种异型高免抗血清。参照经典的鸡血型鉴定方法,初步建立了单倍型鸡血型鉴定红细胞凝集方法,B13、B15、B5、B21、B2和B19的同种异型抗血清血凝效价分别为1∶80、1∶80、1∶40、1∶20、1∶16和1∶32,利用PCR对24只鸡的BF基因外显子2和3进行测序,确定其中22只为B5/B15杂合、2只为B15/B15纯合。利用制备的B15和B5抗血清分别对这24只鸡进行血凝试验,结果表明:B15抗血清与B5/B15个体的血凝效价为0～1∶160,与B15/B15个体的血凝效价分别为1∶80和1∶160;B5抗血清与B5/B15个体的血凝效价为1∶40～1∶80,与B15/B15个体的血凝效价分别为1∶10和1∶20。本研究制备的6种MHC单倍型鸡同种异型抗血清,为鸡的血型鉴定及免疫遗传基因纯合个体的筛选提供了物质基础。Specific pathogen-free （SPF）BW/G （n）chicken lines,termed as BW/G1,BW/G2, BW/G3,BW/G5,BW/G6,BW/G7,were major histocompatibility complex B core areas homozygous chickens,belonging to B13, B15, B2, B5,B21, B19 haplotypes, respectively. In this study, allotype antisera were prepared by immunizing haplotype chickens with peripheral red blood ceils derived from other haplotype chickens. A hemagglutination assay （HA）was established according to modified classic chicken blood type identification method,the antisera titers of B13,B15,B5,B21,B2 and B19 were 1 ： 80,1 ： 80,1 ： 40,1 ： 20,1 ： 16 and 1 ： 32, respectively. While HA titers of B2, B21, B 19 were 0,0,1 ： 80 when detected with B15 antisera, respectively. By directly sequencing the exon 2 and 3 of BF genes, 22 chickens were confirmed as B5/B15 heterozygosities and 2 were B15/B15 homozygosities. 24 chickens were then serotyped using the HA with the B5 and B15 antisera,resulted in B5/B15 individuals were 0 to 1：160 and 1：40 to 1：80 when detected with B15 and B5 antisurum,while two B15/B15 birds were 1：80 and 1：160 each with B15 antisurum,and 1：10 and 1：20 each with B5 antisurum. The prepared allotype antiserum would be beneficial for chicken blood typing and screening of MHC homozygous individuals.

关 键 词：鸡 MHC—B单倍型 B血型 红细胞凝集试验 

分 类 号：S432.41[农业科学—植物病理学]