High sensitivity of one-step real-time reverse transcription quantitative PCR to detect low virus titers in large mosquito pools
Tang, Zhaoyang (Huzhou University. Key Laboratory of Vector Biology and Pathogen Control of Zhejiang Province)
Yamada, Hanano (Vienna International Centre. Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture)
Kraupa, Carina (Vienna International Centre. Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture)
Canic, Sumejja (Vienna International Centre. Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture)
Busquets, Núria (Institut de Recerca i Tecnologia Agroalimentàries. Centre de Recerca en Sanitat Animal)
Talavera Forcades, Sandra (Institut de Recerca i Tecnologia Agroalimentàries. Centre de Recerca en Sanitat Animal)
Jiolle, Davy (Institut de Recherche pour le Développement. Maladies Infectieuses et Vecteurs)
Vreysen, Marc J. B. (Vienna International Centre. Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture)
Bouyer, Jérémy (Vienna International Centre. Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture)
Abd-Alla, Adly M. M. (Vienna International Centre. Joint FAO/IAEA Division of Nuclear Techniques in Food and Agriculture)

Fecha:	2020
Resumen:	Mosquitoes are the deadliest animals in the world. Their ability to carry and spread diseases to humans causes millions of deaths every year. Due to the lack of efficient vaccines, the control of mosquito-borne diseases primarily relies on the management of the vector. Traditional control methods are insufficient to control mosquito populations. The sterile insect technique (SIT) is an additional control method that can be combined with other control tactics to suppress specific mosquito populations. The SIT requires the mass-rearing and release of sterile males with the aim to induce sterility in the wild female population. Samples collected from the environment for laboratory colonization, as well as the released males, should be free from mosquito-borne viruses (MBV). Therefore, efficient detection methods with defined detection limits for MBV are required. Although a one-step reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) method was developed to detect arboviruses in human and mosquito samples, its detection limit in mosquito samples has yet to be defined. We evaluated the detection sensitivity of one step RT-qPCR for targeted arboviruses in large mosquito pools, using pools of non-infected mosquitoes of various sizes (165, 320 and 1600 mosquitoes) containing one infected mosquito body with defined virus titers of chikungunya virus (CHIKV), usutu virus (USUV), West Nile virus (WNV) and Zika virus (ZIKV). CHIK, USUV, ZIKV, and WNV virus were detected in all tested pools using the RT-qPCR assay. Moreover, in the largest mosquito pools (1600 mosquitoes), RT-qPCR was able to detect the targeted viruses using different total RNA quantities (10, 1 and 0. 1 ng per reaction) as a template. Correlating the virus titer with the total RNA quantity allowed the prediction of the maximum number of mosquitoes per pool in which the RT-qPCR can theoretically detect the virus infection. Mosquito-borne viruses can be reliably detected by RT-qPCR assay in pools of mosquitoes exceeding 1000 specimens. This will represent an important step to expand pathogen-free colonies for mass-rearing sterile males for programmes that have a SIT component by reducing the time and the manpower needed to conduct this quality control process.
Ayudas:	European Commission 731060
Nota:	Altres ajuts: China Scholarship Council 201806850127
Derechos:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Lengua:	Anglès
Documento:	Article ; recerca ; Versió publicada
Materia:	Flavivirus ; Arbovirus ; Chikungunya virus (CHIKV) ; Usutu virus (USUV) ; West Nile virus (WNV) ; Zika virus (ZIKV) ; Pool size ; RT-qPCR
Publicado en:	Parasites & vectors, Vol. 13 (september 2020) , ISSN 1756-3305

DOI: 10.1186/s13071-020-04327-4
PMID: 32907625

13 p, 2.4 MB

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