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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 ZohdyComments to Author 
Author affiliations: US President’s Malaria Initiative, Centers for Disease Control and Prevention, Atlanta, Georgia, USA (C. Rafferty, S. Zohdy); Centers for Disease Control and Prevention, Atlanta (C. Rafferty, G. Raise, J. Scaife, S. Zohdy); Kenya Medical Research Institute (KEMRI), Kisumu, Kenya (B. Abongo, S. Omondi, S. Milanoi, M. Muchoki, B. Onyango, E. Ochomo)

Main Article

Figure 4

Validation of colorimetric loop-mediated isothermal amplification assay to detect invasive malaria vector Anopheles stephensi mosquitoes. A) Results using new assay. B) Results using existing An. stephensi PCR (13); yellow boxes indicate An. stephensi products. Conventional PCR resulted in difficult-to-interpret gel bands for An. longipalpis C, Aedes aegypti, and An. coustani samples, similar to An. stephensi samples, and inconsistently produced double bands (positive detection) on sequence-confirmed An. stephensi samples from Kenya. Asterisks in key indicate samples from insectary-reared mosquitoes. Samples 12–23 came from sequence-confirmed field-collected specimens. Sample 25 contained a pool of assorted mosquito DNA species, in which An. stephensi was represented 1:10. For both assays, 1 µL extracted DNA was used. NDC, no DNA template control; UCI, An. stephensi laboratory colony (BEI Resources, https://www.beiresources.org).

Figure 4. Validation of colorimetric loop-mediated isothermal amplification assay to detect invasive malaria vector Anopheles stephensi mosquitoes. A) Results using new assay. B) Results using existing An. stephensi PCR (13); yellow boxes indicate An. stephensi products. Conventional PCR resulted in difficult-to-interpret gel bands for An. longipalpis C, Aedes aegypti, and An. coustani samples, similar to An. stephensi samples, and inconsistently produced double bands (positive detection) on sequence-confirmed An. stephensi samples from Kenya. Asterisks in key indicate samples from insectary-reared mosquitoes. Samples 12–23 came from sequence-confirmed field-collected specimens. Sample 25 contained a pool of assorted mosquito DNA species, in which An. stephensi was represented 1:10. For both assays, 1 µL extracted DNA was used. NDC, no DNA template control; UCI, An. stephensi laboratory colony (BEI Resources, https://www.beiresources.org).

Main Article

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Page created: July 10, 2024
Page updated: July 10, 2024
Page reviewed: July 10, 2024
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