Human short tandem repeat identification using a nanopore-based DNA sequencer: a pilot study

J Hum Genet. 2020 Jan;65(1):21-24. doi: 10.1038/s10038-019-0688-z. Epub 2019 Oct 24.

Abstract

Short tandem repeats (STRs) are repetitive DNA sequences that are highly polymorphic and widely used for personal identification in the field of forensic medicine. The standard method for determining the repeat number of STRs is capillary electrophoresis of PCR products; however, the use of DNA sequencing has increased because it can identify same-sized alleles with nucleotide substitutions (iso-alleles). In this study, we performed human STR genotyping using a portable nanopore-based DNA sequencer, the MinION, and evaluated its performance. Because the sequence quality obtained by MinION is considerably lower than those obtained with other DNA sequencers, we developed an original scoring scheme for judging the genotypes from MinION reads. Analysis of seven human samples for 21-45 STR loci yielded an average of 857 thousand reads per sample, and the accuracy of genotyping and iso-allele identification reached 75.7% and 82%, respectively. Although the accuracy is higher than that reported previously, further improvements are required before this method can be practically applied.

MeSH terms

  • Alleles
  • Female
  • Genotyping Techniques / instrumentation
  • Genotyping Techniques / methods
  • High-Throughput Nucleotide Sequencing / instrumentation
  • High-Throughput Nucleotide Sequencing / methods*
  • Humans
  • Male
  • Microsatellite Repeats / genetics*
  • Nanopore Sequencing / instrumentation
  • Nanopore Sequencing / methods*
  • Pilot Projects
  • Sequence Analysis, DNA / instrumentation
  • Sequence Analysis, DNA / methods*