Development and Validation of Reverse Transcription Loop-Mediated Isothermal Amplification (RT-LAMP) for Rapid Detection of ZIKV in Mosquito Samples from Brazil

Sci Rep. 2019 Mar 14;9(1):4494. doi: 10.1038/s41598-019-40960-5.

Abstract

The rapid spread of Zika virus (ZIKV) represents a global public health problem, especially in areas that harbor several mosquito species responsible for virus transmission, such as Brazil. In these areas, improvement in mosquito control needs to be a top priority, but mosquito viral surveillance occurs inefficiently in ZIKV-endemic countries. Quantitative reverse transcription PCR (qRT-PCR) is the gold standard for molecular diagnostic of ZIKV in both human and mosquito samples. However, the technique presents high cost and limitations for Point-of-care (POC) diagnostics, which hampers its application for a large number of samples in entomological surveillance programs. Here, we developed and validated a one-step reverse transcription LAMP (RT-LAMP) platform for detection of ZIKV in mosquito samples. The RT-LAMP assay was highly specific for ZIKV and up to 10,000 times more sensitive than qRT-PCR. Assay validation was performed using 60 samples from Aedes aegypti and Culex quinquefasciatus mosquitoes collected in Pernambuco State, Brazil, which is at the epicenter of the Zika epidemic. The RT-LAMP had a sensitivity of 100%, specificity of 91.18%, and overall accuracy of 95.24%. Thus, our POC diagnostics is a powerful and inexpensive tool to monitor ZIKV in mosquito populations and will allow developing countries to establish better control strategies for this devastating pathogen.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aedes / virology
  • Animals
  • Brazil
  • Chlorocebus aethiops
  • Culex / virology
  • Culicidae / virology*
  • Female
  • Nucleic Acid Amplification Techniques / methods*
  • Point-of-Care Systems
  • Population Surveillance
  • RNA, Viral / genetics
  • Real-Time Polymerase Chain Reaction / methods*
  • Sensitivity and Specificity
  • Vero Cells
  • Zika Virus / genetics
  • Zika Virus / isolation & purification*
  • Zika Virus Infection / diagnosis*

Substances

  • RNA, Viral