Dengue serotype detection from wild caught mosquito by reverse transcription loop-mediated isothermal amplification

Abstract

Dengue viruses are mainly transmitted by the Aedes aegypti mosquito which grows in breeding containers maintained by rain or human activity. The Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) reaction was found more sensitive and can detect 10-copies of RNA template whereas; real-time PCR (qPCR) had detection limits of 100 copies. To detect the dengue serotypes from wild-caught Aedes aegypti mosquitoes using a molecular technique RT-LAMP, and compare with qPCR to detect the same dengue serotype. This prospective analytical study was conducted at the Department of Virology, BSMMU, Dhaka, Bangladesh from January 2017 to December 2017. RT-LAMP was performed to detect dengue serotypes from wild-caught mosquitoes. Four different visual detection methods of RT-LAMP were conducted simultaneously. To confirm the test result, qPCR was done. Out of the 1348 wild-caught mosquitoes, 217(16.10%) were found positive for the dengue virus by RT-LAMP assay. Among this DEN-2 serotype was by far the highest percentage 184(84.79%) whereas, DEN-1 was 33(15.21%). The RT-LAMP is a relatively inexpensive, rapid, and simple tool for the accurate detection and serotyping of DENV from wild-caught mosquitoes.