High-performance LAMP-based method for human sex identification using Y chromosome-specific genetic markers

Finding fast and cheap strategies for DNA typing and human sample identification is of interest in forensic science. We report a new versatile alternative for molecular sex determination using Y-specific targets (TSPY, TTTY, alphoid regions, and Y-Amelogenin). This system uses an isothermal loop-med...

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Bibliographic Details
Published in:Forensic science international. Genetics supplement series 2022-12, Vol.8, p.225-227
Main Authors: Burgos, Germán, Barrionuevo-Pérez, Katherin, Restrepo, Tomás, Tejera, Eduardo, de Waard, Jacobus H., Garzón-Salazar, Alejandra, Gusmão, Leonor
Format: Article
Language:English
Subjects:
Alphoid repeats
LAMP
Sensitivity
TSPY
TTTY
Y-Amelogenin
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Summary:Finding fast and cheap strategies for DNA typing and human sample identification is of interest in forensic science. We report a new versatile alternative for molecular sex determination using Y-specific targets (TSPY, TTTY, alphoid regions, and Y-Amelogenin). This system uses an isothermal loop-mediated DNA amplification (LAMP) with a set of 6 primers for each target, designed to improve sensibility and specificity, and reducing detection time to only 45 min. Furthermore, detecting the different targets on the Y chromosome either individually or in combination revealed accurate results. Assay sensitivity was determined with a mixture of human female and male DNA at different concentrations to mimic forensic samples. Single primer sets showed high sensitivity at DNA concentrations ranging from 58.6 to 3.7 pg/µL. When a combined primers set was used, sensitivity yielded a detection as low as 0.1 pg/µL of male DNA, making it 10 times more sensitive than qPCR-DNA quantification kits. Finally, high specificity was observed when tested against 6 domestic species.
ISSN:1875-1768
1875-175X
DOI:10.1016/j.fsigss.2022.10.043