A multiplex PCR assay for six Aedini species, including Aedes albopictus
Abstract Background Mosquitoes, as vectors of various human pathogens, are significant drivers of serious human illness. In particular, those species in the Aedini tribe, which typically transmit dengue virus, Chikungunya fever virus, and Zika virus, are increasing their range because of climate cha...
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doaj-f2f0a6b25efe4bd1b81eb757cf83741d2021-08-01T11:28:19ZengBMCParasites & Vectors1756-33052021-07-011411910.1186/s13071-021-04871-7A multiplex PCR assay for six Aedini species, including Aedes albopictusWoo Jun Bang0Min Hyeok Won1Seong Tae Cho2Jihun Ryu3Kwang Shik Choi4School of Life Sciences, Kyungpook National UniversitySchool of Life Sciences, Kyungpook National UniversitySchool of Life Sciences, Kyungpook National UniversitySchool of Life Sciences, Kyungpook National UniversitySchool of Life Sciences, Kyungpook National UniversityAbstract Background Mosquitoes, as vectors of various human pathogens, are significant drivers of serious human illness. In particular, those species in the Aedini tribe, which typically transmit dengue virus, Chikungunya fever virus, and Zika virus, are increasing their range because of climate change and international commerce. In order to evaluate the risk of disease transmission, accurate mosquito species identification and monitoring are needed. The goal of this work was to develop a rapid and simple molecular diagnostic method for six morphologically similar Aedini species (Aedes flavopictus, Aedes albopictus, Ochlerotatus koreicus, Ochlerotatus japonicus, Ochlerotatus togoi and Ochlerotatus hatorii) in Korea. Methods A total of 109 samples were assayed in this study. The internal transcribed spacer 2 (ITS2) regions from all six species were amplified, sequenced and analyzed using Mega 6. Following the identification of regions that were consistently different in terms of sequence between all six species, multiplex primers were designed to amplify these regions to generate species-specific fragments distinguishable by their size. Results Uniquely sized fragments were generated in Ae. flavopictus (495 bp), Ae. albopictus (438 bp), Oc. koreicus (361 bp), Oc. togoi (283 bp), Oc. hatorii (220 bp) and Oc. japonicus (160 bp). Pairwise distance analysis showed that the difference was 35.0 ± 1.5% between Aedes spp. and Ochlerotatus spp., 17.4 ± 0.2% between Ae. albopictus and Ae. flavopictus and 11.1 ± 0.3% between Oc. koreicus and Oc. japonicus. Conclusions In this study, a multiplex PCR assay for six species of the Aedini tribe was developed. This assay is more accurate than morphological identification and will be useful for monitoring and controlling these vector mosquitoes. Graphical Abstracthttps://doi.org/10.1186/s13071-021-04871-7AediniAedes albopictusInternal transcribed spacer 2Multiplex PCR assay |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Woo Jun Bang Min Hyeok Won Seong Tae Cho Jihun Ryu Kwang Shik Choi |
spellingShingle |
Woo Jun Bang Min Hyeok Won Seong Tae Cho Jihun Ryu Kwang Shik Choi A multiplex PCR assay for six Aedini species, including Aedes albopictus Parasites & Vectors Aedini Aedes albopictus Internal transcribed spacer 2 Multiplex PCR assay |
author_facet |
Woo Jun Bang Min Hyeok Won Seong Tae Cho Jihun Ryu Kwang Shik Choi |
author_sort |
Woo Jun Bang |
title |
A multiplex PCR assay for six Aedini species, including Aedes albopictus |
title_short |
A multiplex PCR assay for six Aedini species, including Aedes albopictus |
title_full |
A multiplex PCR assay for six Aedini species, including Aedes albopictus |
title_fullStr |
A multiplex PCR assay for six Aedini species, including Aedes albopictus |
title_full_unstemmed |
A multiplex PCR assay for six Aedini species, including Aedes albopictus |
title_sort |
multiplex pcr assay for six aedini species, including aedes albopictus |
publisher |
BMC |
series |
Parasites & Vectors |
issn |
1756-3305 |
publishDate |
2021-07-01 |
description |
Abstract Background Mosquitoes, as vectors of various human pathogens, are significant drivers of serious human illness. In particular, those species in the Aedini tribe, which typically transmit dengue virus, Chikungunya fever virus, and Zika virus, are increasing their range because of climate change and international commerce. In order to evaluate the risk of disease transmission, accurate mosquito species identification and monitoring are needed. The goal of this work was to develop a rapid and simple molecular diagnostic method for six morphologically similar Aedini species (Aedes flavopictus, Aedes albopictus, Ochlerotatus koreicus, Ochlerotatus japonicus, Ochlerotatus togoi and Ochlerotatus hatorii) in Korea. Methods A total of 109 samples were assayed in this study. The internal transcribed spacer 2 (ITS2) regions from all six species were amplified, sequenced and analyzed using Mega 6. Following the identification of regions that were consistently different in terms of sequence between all six species, multiplex primers were designed to amplify these regions to generate species-specific fragments distinguishable by their size. Results Uniquely sized fragments were generated in Ae. flavopictus (495 bp), Ae. albopictus (438 bp), Oc. koreicus (361 bp), Oc. togoi (283 bp), Oc. hatorii (220 bp) and Oc. japonicus (160 bp). Pairwise distance analysis showed that the difference was 35.0 ± 1.5% between Aedes spp. and Ochlerotatus spp., 17.4 ± 0.2% between Ae. albopictus and Ae. flavopictus and 11.1 ± 0.3% between Oc. koreicus and Oc. japonicus. Conclusions In this study, a multiplex PCR assay for six species of the Aedini tribe was developed. This assay is more accurate than morphological identification and will be useful for monitoring and controlling these vector mosquitoes. Graphical Abstract |
topic |
Aedini Aedes albopictus Internal transcribed spacer 2 Multiplex PCR assay |
url |
https://doi.org/10.1186/s13071-021-04871-7 |
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