In this article, we describe an in-house polymerase chain reaction-sequence specific priming (PCR-SSP) assay designed for undergraduate medical students as part of the experimental pathogen biology and immunology (EPBI) course. It screens human leukocyte antigen (HLA)-DR2 allotype from genomic DNA samples using a rapid and single-tube PCR technique, yielding definitive typing result without conventional post-amplification step like probing or Sanger sequencing. This laboratory exercise offers the undergraduate medical students an opportunity to learn about current molecular biology techniques in HLA genotyping with limited effort and cost, in addition to a better understanding of concepts presented in the classroom lectures. Upon completing this experiment module, the students show statistically significant improvement in several key indexes, such as the knowledge about the mainstream HLA DNA typing techniques, awareness of the relevance of this knowledge for their future scientific research, immunogenetics-related basic laboratory skills they acquire, and interest and desire for mastering this assay (all p < .05). This easy to implement set of experiments is composed of a two-session lab module occupying eight teaching hours, and has been run successfully in our laboratory.

译文

在本文中,我们描述了一种内部聚合酶链反应-序列特异性引发 (pcr-ssp) 测定法,该测定法是为本科生设计的实验病原体生物学和免疫学 (EPBI) 课程的一部分。它使用快速和单管PCR技术从基因组DNA样品中筛选人类白细胞抗原 (HLA)-DR2同种异型,无需常规的扩增后步骤 (如探测或Sanger测序) 即可产生确定的分型结果。这个实验室练习为本科生提供了一个机会,以有限的努力和成本来学习HLA基因分型中的当前分子生物学技术,此外,还可以更好地理解课堂讲座中提出的概念。完成本实验模块后,学生在几个关键指标上显示出统计学上的显着改善,例如有关主流HLA DNA分型技术的知识,对这些知识与未来科学研究的相关性的认识,与免疫遗传学相关的基本实验室技能,以及对掌握这种测定的兴趣和渴望 (所有p  < .05)。这组易于实现的实验由一个两阶段的实验室模块组成,该模块占用了八个教学小时,并已在我们的实验室中成功运行。

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