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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

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 × 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).

Main Article

Page created: July 10, 2024
Page updated: August 20, 2024
Page reviewed: August 20, 2024
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