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

Moufid Mhamadi (1) , Giulia Mencattelli (1) , Alioune Gaye (2) , El Hadji Ndiaye (2) , Aïssatou Aïcha Sow (1) , Martin Faye (1) , Marie Henriette Dior Ndione (1) , Moussa Moïse Diagne (1) , Moundhir Mhamadi (3) , Ousmane Faye (1) , Manfred Weidmann (4) , Oumar Faye (1) , Mawlouth Diallo (3) , Cheikh Tidiane Diagne (1, 3)

1 Département de virologie - Virology Department [Dakar]
2 Pôle Zoologie médicale [Dakar]
3 Diatropix [Dakar, Sénégal]
4 MHB Theodor Fontane - Medizinische Hochschule Brandenburg Theodor Fontane / Brandenburg Medical School Theodor-Fontane

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.

Mots clés: direct RT-qPCR supernatant arbovirus infected mosquitoes field diagnosis; direct RT-qPCR; supernatant; arbovirus; infected mosquitoes; field diagnosis

Domaines: Sciences du Vivant [q-bio]

Moufid Mhamadi, Giulia Mencattelli, Alioune Gaye, El Hadji Ndiaye, Aïssatou Aïcha Sow, et al.. Rapid On-Site Detection of Arboviruses by a Direct RT-qPCR Assay. Biosensors, 2023, 13 (12), pp.1035. ⟨10.3390/bios13121035⟩. ⟨hal-04569994⟩