Development of LAMP-colorimetric assay for detecting hemoglobin Constant Spring and hemoglobin Pakse' v1
Wittaya Jomoui · 2025
A novel molecular detection method based on a colorimetric LAMP with phenol red assay was developed to detect Hb CS and Hb PS for the first time. All LAMP primer sets and probe in this study were designed using Primer Explorer V5 software (http://primerexplorer.jp /lampv5e/index.html). The common primer set including the outer primers (F3 and B3) and the inner primers (FIP and BIP) was shown in the Table. Furthermore, two loop probe (LP) used in this study were designed for specific with Hb CS (LP-CS) and Hb PS (LP-PS) as also shown in Table . These primers set and probes were protected according to petty patent submission number 2203001286 (Thailand). The developed LAMP colorimetric phenol red assay was performed with reaction mixture and temperature as detailed. The LAMP mixture was incubated at an isothermal temperature of 65 °C for 35 minutes. The positive result was observed as yellow color by the naked eye compared with negative with pink color.