Flow Chart HLA-DQA1 Genotyping and Its Application to a Forensic Case

Abstract

The detection of genetic polymorphism has become increasingly important in forensic science as well as in medical genetics. In this report, we describe a systematic flow chart system for HLA-DQA1 genotyping by an improved PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphism) method coupled with the PRSM (PCR-mediated restriction site modification) method. This flow chart typing system can easily discriminate between a total of eight reported DQA1 alleles commonly found in Chinese. We have applied this flow chart typing system in a forensic case as well as in the determination of the frequencies of the eight DQA1 alleles in 121 unrelated Taiwan Chinese subjects. Our results show that the flow chart DQA1 genotyping is a simple, fast, and accurate system which, in the future, may be considered as an alternative method for routine individual identification in forensic casework, and for paternity testing and tissue typing in medical genetics.

Fidelity of polymerase chain reaction-direct sequencing analysis of damaged forensic samples

“…A 373A DNA Sequencer (Applied Biosystems, Foster City, CA, USA) was employed for sequence gel analysis and the traces were always visually checked independently by two operators. Allelic designation was achieved by comparing our sequencing data with the published sequences of the alleles at HLA DQA1 locus [35].…”

Forensic Science

“…The PM system has been validated for forensic case work (736). A systematic flow chart system for HLA-DQA1 genotyping has been proposed (737). Typing protocols are recommended for obtaining reliable results with the PM system (738).…”

Time‐Resolved Fluorometry Based Sandwich Hybridisation Assay for HLA‐DQA1 Typing

“…Various modifications of the method such as reverse dot blot and replacement of radioactive isotopes by alternative labels like digoxigenin have been made. Alternatives presented later on include PCR-RFLP (polymerase chain reaction restriction fragment length polymorphism) [18,25,26], sequence-specific amplification [1,2,5,7,23,24] and finally direct sequencing [29] of the amplification product. The final choice of the method for HLA typing depends on several aspects such as cost of the assay (equipment and reagents required), number of samples to be screened and assay related characteristics (specificity, sensitivity, reproducibility, degree of automation etc.…”