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Volume 30, Number 9—September 2024
Research
Loop-Mediated Isothermal Amplification Assay to Detect Invasive Malaria Vector Anopheles stephensi Mosquitoes
Table 4
Assessment of assay sensitivity for development of colorimetric loop-mediated isothermal amplification assay to detect invasive malaria vector Anopheles stephensi mosquitoes*
Template dilution | Template concentration, ng | UCI |
---|---|---|
NTC | NA | 3/3 negative |
1 | 311.6 | 3/3 positive |
10 | 31.16 | 3/3 positive |
1 × 102 | 3.12 | 3/3 positive |
1 × 103 | 0.312 | 3/3 positive |
1 × 104 | 0.031 | 3/3 positive |
1 × 105 | 0.003 | 3/3 positive |
1 × 106 | 0.0003 | 3/3 positive |
1 × 107 | 0.00003 | 1/3 positive, 2/3 negative |
*Assessment was done using a serial (1:10) dilution from DNA extract containing 311.6 ng of Anopheles stephensi (UCI) whole mosquito DNA and resulted in 100% positive color change down to 0.0003 ng. Each dilution test was performed in triplicate. Color change at 1 x 107 was inconsistent, and samples with DNA concentrations <0.0003 were less likely to result in a positive color change and did not display as clear pink or yellow. NA, not applicable; NTC, no template control; UCI, An. stephensi laboratory colony (BEI Resources, https://www.beiresources.org).