A laboratory practice that uses the polymerase chain reaction-sequence specific priming technique to rapidly screen for HLA-DR2 allotype from germline DNA in immunology course for undergraduate medical students

Biochem Mol Biol Educ. 2020 Mar;48(2):175-180. doi: 10.1002/bmb.21322. Epub 2019 Dec 3.

Abstract

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.

Keywords: PCR-SSP; human leukocyte antigen; immunogenetics; laboratory exercises.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alleles
  • Curriculum*
  • DNA / genetics*
  • Education, Medical, Undergraduate
  • Germ Cells / metabolism*
  • HLA-DR2 Antigen / genetics*
  • Humans
  • Laboratories*
  • Polymerase Chain Reaction*
  • Sequence Analysis, DNA*
  • Students, Medical

Substances

  • HLA-DR2 Antigen
  • DNA