Identification of human DNA by loop-mediated isothermal amplification (LAMP) technique combined with white ring precipitation of Cu(OH)2

Abstract

Loop-mediated isothermal amplification (LAMP) analysis has been developed for human identification. DNA templates of 8 sample species including Homo sapiens (human) were subjected to LAMP for apolipoprotein L1 gene (ApoL1) amplification. After DNA amplification, gel electrophoresis was performed. Furthermore, CuSO4 was added in to the LAMP product for staining visible to the naked eye. The LAMP products were only present with human DNA. The limit of detection was reproducibly as low as 10pg of genomic DNA. Moreover, by adding CuSO4 to the LAMP product, the Cu(OH)2 precipitate, which formed a ring-shaped deposit, was not seen with the human DNA, but was seen with the LAMP negative samples. Our findings show the LAMP technique to be a powerful method for identifying the presence of human DNA. The addition of CuSO4 to the LAMP product is an alternative that can be used in field studies, and does not require access to gel-electrophoresis.

Keywords

• human identification
• loop-mediated isothermal amplification
• LAMP
• APOL1