LAMP Detection of Virus-Derived DNA of Zika Virus in Vector Mosquito

Manabu Ote,Hiroka Aonuma,Hirotaka Kanuka,Chun-Hong Chen,Jian-Chiuan Li,Shigeru Tajima,Itoe Iizuka,Masayuki Saijo

doi:10.3389/fitd.2022.759375

Abstract

Detection of infectious viruses in mosquitoes is one of the prerequisite measures to monitor the prevalence of vector-borne viral diseases. Determining which mosquitoes are currently infected with arboviruses such as Zika, dengue, and chikungunya virus is not yet practical in endemic areas due to multiple causes including the difficulty of dealing with the virus’ unstable RNA. In this study, instead of handling viral RNA, virus-derived DNA (vDNA) was introduced as a target template for nucleic acid amplification. In combination with loop-mediated isothermal amplification (LAMP), we examined a LAMP-based vDNA detection assay (vDNA-LAMP) targeting Zika virus (ZIKV). The vDNA-LAMP reaction amplifying part of the NS3 region of ZIKV successfully detected its vDNA from crude DNA purified from artificially infected cultured cells and Aedes mosquitoes. This rapid, simple, and versatile method may provide a promising field-surveillance method for arbovirus circulation via vector mosquitoes.

Highlights

Zika virus (ZIKV) is one of the family Flaviviridae, genus flavivirus, which is closely related to dengue virus
We demonstrated that loop-mediated isothermal amplification (LAMP) detected virus-derived DNA (vDNA) of ZIKV from both cultured cells and vector mosquitoes, making LAMP targeting vDNA a candidate method applicable for on-site surveillance of mosquitoes carrying arboviruses
The target region of ZIKV was amplified from all cDNA samples, suggesting that the primer set can be applicable to vDNA-LAMP (Supplementary Figure 1B)

Summary

Introduction

Zika virus (ZIKV) is one of the family Flaviviridae, genus flavivirus, which is closely related to dengue virus. Infection with ZIKV in humans passes with an asymptomatic course in most cases. ZIKV infection during pregnancy causes microcephaly and adverse fetal outcomes [1]. For which the natural vertebrate host is humans, ZIKV outbreaks are often a result of spillover from sylvatic reservoirs [2]. ZIKV transmission is expected to be induced primarily by blood feeding of Aedes mosquitoes including Aedes aegypti and Ae. albopictus. Vertical transmission of ZIKV in Aedes mosquitoes, like other flaviviruses including dengue, has been demonstrated by laboratory studies [3,4,5]. It has been reported that wild Ae. aegypti larvae carried ZIKV, presenting natural vertical transmission [6]. Hindering the authentic and alternative ZIKV transmission cycles via mosquitoes may mitigate this increasing health burden

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Discussion

Conclusion