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Methodology
Kwang Shik Choi
Kyungpook National University, Daehak-ro 80, Daegu
Corresponding Author
License:
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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 prevent mosquito-borne disease, 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
In total, 109 samples were used in this study. The internal transcribed spacer 2 (ITS2) regions from all six species were amplified, sequenced and analyzed using Mega 6. Once regions that were consistently different in sequence between all six species were identified, multiplex primers were designed to amplify them to generate species-specific fragments distinguishable by their size.
Results
Uniquely sized fragments were generated in Aedes flavopictus (495bp), Aedes albopictus (438bp), Ochlerotatus koreicus (361bp), Ochlerotatus togoi (283bp), Ochlerotatus hatorii (220bp), and Ochlerotatus japonicus (160bp). Pairwise distance analysis showed that the difference was 35.0±1.5% between Aedes spp. and Ochlerotatus spp., 17.4±0.2% between Aedes albopictus and Aedes flavopictus, and 11.1±0.3% between Ochlerotatus koreicus and Ochlerotatus 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.
Keywords
Aedini species, Aedes albopictus, Internal transcribed spacer 2(ITS2), multiplex PCR assay
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Peer Review Timeline
CURRENT STATUS: Published
View the published article at Parasites & Vectors.
Version 1
Posted 15 Dec, 2020
No community comments so far
Editorial decision: Major Revision
On 27 Feb, 2021
Review #2 received
Received 02 Feb, 2021
Reviewer #2 agreed
On 21 Jan, 2021
Review #1 received
Received 21 Jan, 2021
Reviewer #1 agreed
On 10 Jan, 2021
Reviewers invited
Invitations sent on 10 Jan, 2021
Editor invited
On 11 Dec, 2020
Editor assigned
On 11 Dec, 2020
Submission checks complete
On 11 Dec, 2020
First submitted
On 10 Dec, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Parasitology
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A new preprint design is coming!
We have improved readability, clarity, accessibility, and opportunities for engagement.
See the published version of this preprint at Parasites & Vectors.
This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Learn more about In Review
Methodology
Kwang Shik Choi
Kyungpook National University, Daehak-ro 80, Daegu
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
Peer Review Timeline
CURRENT STATUS: Published
View the published article at Parasites & Vectors.
Version 1
Posted 15 Dec, 2020
No community comments so far
Editorial decision: Major Revision
On 27 Feb, 2021
Review #2 received
Received 02 Feb, 2021
Reviewer #2 agreed
On 21 Jan, 2021
Review #1 received
Received 21 Jan, 2021
Reviewer #1 agreed
On 10 Jan, 2021
Reviewers invited
Invitations sent on 10 Jan, 2021
Editor invited
On 11 Dec, 2020
Editor assigned
On 11 Dec, 2020
Submission checks complete
On 11 Dec, 2020
First submitted
On 10 Dec, 2020
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
Subject Areas
Parasitology
License:
This work is licensed under a CC BY 4.0 License. Read Full License
View the published article at Parasites & Vectors.
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 prevent mosquito-borne disease, 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
In total, 109 samples were used in this study. The internal transcribed spacer 2 (ITS2) regions from all six species were amplified, sequenced and analyzed using Mega 6. Once regions that were consistently different in sequence between all six species were identified, multiplex primers were designed to amplify them to generate species-specific fragments distinguishable by their size.
Results
Uniquely sized fragments were generated in Aedes flavopictus (495bp), Aedes albopictus (438bp), Ochlerotatus koreicus (361bp), Ochlerotatus togoi (283bp), Ochlerotatus hatorii (220bp), and Ochlerotatus japonicus (160bp). Pairwise distance analysis showed that the difference was 35.0±1.5% between Aedes spp. and Ochlerotatus spp., 17.4±0.2% between Aedes albopictus and Aedes flavopictus, and 11.1±0.3% between Ochlerotatus koreicus and Ochlerotatus 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.
Figure 1
Figure 2
Figure 3
Figure 4
This is a list of supplementary files associated with this preprint. Click to download.
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