The classical class I and class II molecules of the major histocompatibility complex (MHC) play crucial roles in immune responses to infectious pathogens and vaccines as well as being important for autoimmunity, allergy, cancer and reproduction. These classical MHC genes are the most polymorphic known, with roughly 10,000 alleles in humans. In chickens, the MHC (also known as the BF-BL region) determines decisive resistance and susceptibility to infectious pathogens, but relatively few MHC alleles and haplotypes have been described in any detail. We describe a typing protocol for classical chicken class I (BF) and class II B (BLB) genes based on a hybridization method called reference strand-mediated conformational analysis (RSCA). We optimize the various steps, validate the analysis using well-characterized chicken MHC haplotypes, apply the system to type some experimental lines and discover a new chicken class I allele. This work establishes a basis for typing the MHC genes of chickens worldwide and provides an opportunity to correlate with microsatellite and with single nucleotide polymorphism (SNP) typing for approaches involving imputation.

中文翻译：

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杂交技术的开发和优化，用于对鸡MHC的经典I类和II B类基因进行分型。

主要组织相容性复合物（MHC）的经典I类和II类分子在对传染性病原体和疫苗的免疫反应中起着至关重要的作用，并且对自身免疫，过敏，癌症和生殖也很重要。这些经典的MHC基因是已知最多态的，在人类中大约有10,000个等位基因。在鸡中，MHC（也称为BF-BL区）决定了对感染性病原体的决定性抗药性和易感性，但是相对详细地描述了相对较少的MHC等位基因和单倍型。我们描述了一种基于称为参考链介导的构象分析（RSCA）的杂交方法的经典I类（BF）和II B类（BLB）基因鸡的打字协议。我们优化了各个步骤，使用特征明确的鸡MHC单倍型验证了分析，应用系统键入一些实验系，并发现一个新的I类鸡等位基因。这项工作为全世界鸡的MHC基因分型奠定了基础，并为与微卫星和单核苷酸多态性（SNP）分型相关的插补方法提供了机会。