Volume 30, Number 9—September 2024
Research
Loop-Mediated Isothermal Amplification Assay to Detect Invasive Malaria Vector Anopheles stephensi Mosquitoes
Cristina Rafferty, Gloria Raise, JeNyiah Scaife, Bernard Abongo, Seline Omondi, Sylvia Milanoi, Margaret Muchoki, Brenda Onyango, Eric Ochomo, and Sarah Zohdy
Table 6
Testing of wild Anopheles spp. mosquitoes collected in Marsabit, Kenya, in 2022 and 2023 that failed to amplify during routine species assays as part of development of colorimetric LAMP assay to detect invasive malaria vector Anopheles stephensi mosquitoes*
| Site | Collection method | Date of collection | Sequence-confirmed species | Colorimetric LAMP assay result |
|---|---|---|---|---|
| Marsabit | Light trap | 2023 Feb | An. gambiae senus lato | Negative (0/2) |
| Marsabit | Light trap | 2023 Feb | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 Feb | An. dthali | Negative (0/2) |
| Marsabit | Larvae | 2023 Feb | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 Feb | An. dthali | Negative (0/2) |
| Marsabit | Larvae | 2023 Feb | Culex peresiguus | Negative (0/2) |
| Marsabit | Larvae | 2023 Feb | An. dthali | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | NA | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | An. stephensi | Positive (2/2) |
| Marsabit | Adults | 2022 Dec | NA | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | An. dthali | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | NA | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | NA | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | NA | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | An. dthali | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | An. stephensi | Positive (2/2) |
| Marsabit | Adults | 2022 Dec | NA | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | NA | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | Other | Negative (0/2) |
| Marsabit | Adults | 2022 Dec | NA | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. pretoriensis | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. stephensi | Positive (2/2) |
| Marsabit | Larvae | 2023 May | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. pretoriensis | Negative (0/2) |
| Marsabit | Larvae | 2023 May | NA | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. stephensi | Positive (2/2) |
| Marsabit | Larvae | 2023 May | An. stephensi | Positive (2/2) |
| Marsabit | Larvae | 2023 May | Other | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. stephensi | Positive (2/2) |
| Marsabit | Larvae | 2023 May | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. stephensi | Positive (2/2) |
| Marsabit | Larvae | 2023 May | NA | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. pretoriensis | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. pretoriensis | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. pretoriensis | Negative (0/2) |
| Marsabit | Larvae | 2023 May | NA | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. stephensi | Positive (2/2) |
| Marsabit | Larvae | 2023 May | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. stephensi | Positive (2/2) |
| Marsabit | Larvae | 2023 May | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. pretoriensis | Negative (0/2) |
| Marsabit | Larvae | 2023 May | Other | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 May | NA | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. gambiae s.l. | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. pretoriensis | Negative (0/2) |
| Marsabit | Larvae | 2023 May | An. pretoriensis | Negative (0/2) |
*Samples were sequenced using cytochrome c oxidase subunit I sequence testing to confirm species identification, and then DNA extract from the samples was run in duplicate through the colorimetric LAMP assay. Results showed 100% concurrence between the LAMP assay results and Sanger sequencing in determining which specimens were An. stephensi and which were not. LAMP, loop-mediated isothermal amplification; NA, no amplification occurred; other, other (non-Anopheles) mosquito species.
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