Abstract

This study aimed to apply the polymerase chain reaction-nucleic acid lateral flow immunoassay (PCR-NALFIA) and AS-PCR-NALFIA assays to detect human DNA and determine human gender based on the human sex-determining region Y (SRY) gene and to detect G1138A mutation in the fibroblast growth factor receptor 3 (FGFR3) gene causing achondroplasia (ACH), respectively. PCR primers were designed to target the human SRY gene. Furthermore, AS-PCR primers were modified to specify the G1138A mutation of the FGFR3 gene. Primers were labeled with biotin at 5′ end of forward primer and FAM at the 5′ end of reverse primer to detect the target gene, followed by detection of the double labeled amplicon by NALFIA system. The result from PCR could be interpreted within 5 min by the naked eye. The limit of detection of these assays was up to 1 ng of DNA template. These results were also target gene-specific and no cross-contamination with non-target genes was observed. It is an innovative new tool that shows potential for DNA identification and is easily adaptable to other target detection.

Key words: FGFR3, G1138A, Nucleic acid lateral flow immunoassay (NALFIA), PCR, SRY