Christ R, Hunsmann G, Sauermann U
German Primate Center, Department of Virology and Immunology, Göttingen, FRG.
Tissue Antigens. 1994 Oct;44(4):241-7. doi: 10.1111/j.1399-0039.1994.tb02389.x.
In spite of the widespread use of rhesus monkeys (Macaca mulatta) in biomedical research, MHC typing of this species is not yet routine. Since suitable antibodies are lacking, serological typing of Mamu-DQA1 is not feasible. We developed a typing protocol for MhcMamu-DQA1 from published sequences of the second exon of Mamu-DQA1. This protocol is based on the amplification of the second exon of Mamu-DQA1 with one specific primer pair followed by a "diagnostic" digestion of the PCR products with, at most, 5 different restriction endonucleases. This modified PCR-RFLP permits the rapid identification of 11 out of 13 Mamu-DQA1 alleles in homozygous and heterozygous combinations. The protocol was validated by cloning and sequencing the PCR-products of several animals of different geographic origin. In addition, an as yet unknown allele was detected by PCR-RFLP and was subsequently cloned and its nucleotide sequence determined. All examined sequences except the new allele were identical to those previously published. Therefore, we assume that many of the Mamu-DQA1 alleles of rhesus monkeys have been identified molecularly and that the typing technique presented here can reliably identify Mamu-DQA1 alleles.
尽管恒河猴（猕猴）在生物医学研究中被广泛使用，但该物种的MHC分型尚未成为常规操作。由于缺乏合适的抗体，对Mamu - DQA1进行血清学分型是不可行的。我们根据已发表的Mamu - DQA1第二外显子序列开发了一种MhcMamu - DQA1分型方案。该方案基于使用一对特异性引物对扩增Mamu - DQA1的第二外显子，然后用最多5种不同的限制性内切酶对PCR产物进行“诊断性”消化。这种改良的PCR - RFLP方法能够快速鉴定纯合和杂合组合中13个Mamu - DQA1等位基因中的11个。通过对几只不同地理来源动物的PCR产物进行克隆和测序，验证了该方案。此外，通过PCR - RFLP检测到一个未知等位基因，随后对其进行克隆并确定了核苷酸序列。除新等位基因外，所有检测序列均与先前发表的序列相同。因此，我们认为恒河猴的许多Mamu - DQA1等位基因已通过分子方法鉴定出来，并且这里介绍的分型技术能够可靠地鉴定Mamu - DQA1等位基因。
