Rapid On-Site Detection of Arboviruses by a Direct RT-qPCR Assay

Biosensors (Basel). 2023 Dec 16;13(12):1035. doi: 10.3390/bios13121035.

Abstract

Arthropod-borne diseases currently constitute a source of major health concerns worldwide. They account for about 50% of global infectious diseases and cause nearly 700,000 deaths every year. Their rapid increase and spread constitute a huge challenge for public health, highlighting the need for early detection during epidemics, to curtail the virus spread, and to enhance outbreak management. Here, we compared a standard quantitative polymerase chain reaction (RT-qPCR) and a direct RT-qPCR assay for the detection of Zika (ZIKV), Chikungunya (CHIKV), and Rift Valley Fever (RVFV) viruses from experimentally infected-mosquitoes. The direct RT-qPCR could be completed within 1.5 h and required 1 µL of viral supernatant from homogenized mosquito body pools. Results showed that the direct RT-qPCR can detect 85.71%, 89%, and 100% of CHIKV, RVFV, and ZIKV samples by direct amplifications compared to the standard method. The use of 1:10 diluted supernatant is suggested for CHIKV and RVFV direct RT-qPCR. Despite a slight drop in sensitivity for direct PCR, our technique is more affordable, less time-consuming, and provides a better option for qualitative field diagnosis during outbreak management. It represents an alternative when extraction and purification steps are not possible because of insufficient sample volume or biosecurity issues.

Keywords: arbovirus; direct RT-qPCR; field diagnosis; infected mosquitoes; supernatant.

MeSH terms

  • Animals
  • Arboviruses*
  • Chikungunya Fever* / diagnosis
  • Chikungunya Fever* / epidemiology
  • Chikungunya virus* / genetics
  • Culicidae*
  • Dengue Virus*
  • Zika Virus Infection* / diagnosis
  • Zika Virus* / genetics