Development of Loop mediated isothermal amplification (LAMP) of SRY gene in  human blood samples for sex determination

Panan Kanchanaphum; Titinan Sarataphan; Wirat Thirasan; Gun Anatasomboon

Authors

Panan Kanchanaphum Biochemistry Unit, Faculty of Science, Rangsit University, Patumthani 12000, Thailand
Titinan Sarataphan Biochemistry Unit, Faculty of Science, Rangsit University, Patumthani 12000, Thailand
Wirat Thirasan Biochemistry Unit, Faculty of Science, Rangsit University, Patumthani 12000, Thailand
Gun Anatasomboon Anatomy Unit, Faculty of Science, Rangsit University, Patumthani 12000, Thailand

Keywords:

sex determination, Loop-mediated isothermal amplification, LAMP, SRY gene, blood stained sample

Abstract

Sex determination of the source of forensic DNA evidence is an importance step in crime investigation. LAMP method is a novel molecular technique for determine the gender. SRY gene is male specific gene and used to indicate male genotype. In this study, we develop the new method to determine gender by using Loop mediated isothermal amplification (LAMP) technique from fresh blood sample and blood stained sample. DNA templates from fresh blood samples were extracted and added to a LAMP reaction. In addition, DNA extracted from blood stained sample in filter papers or cloths were used for LAMP reaction. Blood stained samples in filter and cloths were stored at room temperature for 0, 1, 7, 14, 30, and 45 days. Sex determination was achieved with LAMP technique for fresh blood samples and both blood stained samples on filter papers and cloths. LAMP products showed smear bands only in the male sample. LAMP products of samples from blood stained on filter papers kept at room temperature for up to 7 days were still detected while those of blood samples stained on cloth were detected after being kept at room temperature for up to 30 days. We have developed the LAMP method to amplify the SRY gene from fresh blood samples and blood stained samples on filter paper and cloth to determine the gender. Blood samples stained on cloths are more stable than those on filter papers. The authors of this study are confident that this finding will lead to a new practical application in the field of forensic and crime investigations.

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