A laboratory practice that uses the polymerase chain reaction‐sequence specific priming technique to rapidly screen for HLA‐DR2 allotype from germline DNA in …

LX Li, W Tian, J Zhang - Biochemistry and Molecular Biology …, 2020 - Wiley Online Library
LX Li, W Tian, J Zhang
Biochemistry and Molecular Biology Education, 2020Wiley Online Library
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 …
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.
Wiley Online Library