A standardized protocol for the detection of arboviruses in different Aedes mosquito species in North Borneo Sabah, Malaysia
Arboviruses, including dengue, Zika and chikungunya viruses, are mainly transmitted by Aedes mosquitoes and pose a threat to public health. The viruses are transmitted by the primary vector, Aedes aegypti, which is more commonly found in urban environments. However, with increasing urbanization, the...
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Elsevier
2025-12-01
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| Series: | MethodsX |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016125003930 |
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| author | Adlar Ryan Ngiam Saiful Anuar Ju Ahmad Mohd Farid Alias Mohd Arshil Moideen Norsyahida Sulaiman Song-Quan Ong Eric Chong Tzyy Jiann Ping-Chin Lee Sylvia Daim Jodi M. Fiorenzano Noel Cote Andrew G. Letizia |
| author_facet | Adlar Ryan Ngiam Saiful Anuar Ju Ahmad Mohd Farid Alias Mohd Arshil Moideen Norsyahida Sulaiman Song-Quan Ong Eric Chong Tzyy Jiann Ping-Chin Lee Sylvia Daim Jodi M. Fiorenzano Noel Cote Andrew G. Letizia |
| author_sort | Adlar Ryan Ngiam |
| collection | DOAJ |
| description | Arboviruses, including dengue, Zika and chikungunya viruses, are mainly transmitted by Aedes mosquitoes and pose a threat to public health. The viruses are transmitted by the primary vector, Aedes aegypti, which is more commonly found in urban environments. However, with increasing urbanization, the overlap of rural and forested areas where different Aedes species are found could also contribute to transmission. Nevertheless, most extraction methods focus on human blood samples or on Ae. aegypti, which limits standardization of virus detection in a variety of less common Aedes populations, especially sylvatic species.In this study, we demonstrated a standardized protocol for extracting sufficient amounts of RNA for detection from a single mosquito sample. We validated the protocol by extracting arboviruses from six different Aedes species collected in the field in Sabah, Malaysia: Ae. aegypti, Ae. albopictus, Ae. poecilus, Ae. butleri, Ae. niveus, and Ae. vexans. Multiplex real-time PCR detection yielded consistent cycle threshold (Ct) values across species (range: 18.9–40.3), with a positivity cut-off of Ct < 41. Our results show that this protocol improves current practice by extending the target sample to different Aedes mosquito species, ultimately contributing to more efficient virus detection and supporting more comprehensive surveillance, even in ecologically diverse environments. |
| format | Article |
| id | doaj-art-2ee4b6889c56421dbd6592c7a0588af9 |
| institution | Kabale University |
| issn | 2215-0161 |
| language | English |
| publishDate | 2025-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj-art-2ee4b6889c56421dbd6592c7a0588af92025-08-20T03:38:58ZengElsevierMethodsX2215-01612025-12-011510354910.1016/j.mex.2025.103549A standardized protocol for the detection of arboviruses in different Aedes mosquito species in North Borneo Sabah, MalaysiaAdlar Ryan Ngiam0Saiful Anuar Ju Ahmad1Mohd Farid Alias2Mohd Arshil Moideen3Norsyahida Sulaiman4Song-Quan Ong5Eric Chong Tzyy Jiann6Ping-Chin Lee7Sylvia Daim8Jodi M. Fiorenzano9Noel Cote10Andrew G. Letizia11Faculty of Science and Natural Resources, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, MalaysiaMalaysian Armed Forces (MAF), MalaysiaMalaysian Armed Forces (MAF), MalaysiaFaculty of Medicine & Defence Health, National Defence University Malaysia, MalaysiaFaculty of Science and Natural Resources, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, MalaysiaInstitute for Tropical Biology and Conservation, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah Malaysia; Vysnova Partners, LLC, Alexandria, VA 22314, USA; Corresponding authors at: Institute for Tropical Biology and Conservation, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, Malaysia.Biotechnology Research Institute, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, MalaysiaFaculty of Science and Natural Resources, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, Malaysia; Biotechnology Research Institute, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, Malaysia; Vysnova Partners, LLC, Alexandria, VA 22314, USA; Corresponding authors at: Institute for Tropical Biology and Conservation, Universiti Malaysia Sabah, Jalan UMS, 88400 Kota Kinabalu, Sabah, Malaysia.Faculty of Medicine and Health Sciences, Universiti Malaysia Sabah, Kota Kinabalu, Sabah, MalaysiaUS. Naval Medical Research Unit INDO PACIFIC, SingaporeUS. Naval Medical Research Unit INDO PACIFIC, SingaporeUS. Naval Medical Research Unit INDO PACIFIC, SingaporeArboviruses, including dengue, Zika and chikungunya viruses, are mainly transmitted by Aedes mosquitoes and pose a threat to public health. The viruses are transmitted by the primary vector, Aedes aegypti, which is more commonly found in urban environments. However, with increasing urbanization, the overlap of rural and forested areas where different Aedes species are found could also contribute to transmission. Nevertheless, most extraction methods focus on human blood samples or on Ae. aegypti, which limits standardization of virus detection in a variety of less common Aedes populations, especially sylvatic species.In this study, we demonstrated a standardized protocol for extracting sufficient amounts of RNA for detection from a single mosquito sample. We validated the protocol by extracting arboviruses from six different Aedes species collected in the field in Sabah, Malaysia: Ae. aegypti, Ae. albopictus, Ae. poecilus, Ae. butleri, Ae. niveus, and Ae. vexans. Multiplex real-time PCR detection yielded consistent cycle threshold (Ct) values across species (range: 18.9–40.3), with a positivity cut-off of Ct < 41. Our results show that this protocol improves current practice by extending the target sample to different Aedes mosquito species, ultimately contributing to more efficient virus detection and supporting more comprehensive surveillance, even in ecologically diverse environments.http://www.sciencedirect.com/science/article/pii/S2215016125003930RNA extractionAedes mosquitoesMultiplex real-time PCRDisease surveillanceDengueZika |
| spellingShingle | Adlar Ryan Ngiam Saiful Anuar Ju Ahmad Mohd Farid Alias Mohd Arshil Moideen Norsyahida Sulaiman Song-Quan Ong Eric Chong Tzyy Jiann Ping-Chin Lee Sylvia Daim Jodi M. Fiorenzano Noel Cote Andrew G. Letizia A standardized protocol for the detection of arboviruses in different Aedes mosquito species in North Borneo Sabah, Malaysia MethodsX RNA extraction Aedes mosquitoes Multiplex real-time PCR Disease surveillance Dengue Zika |
| title | A standardized protocol for the detection of arboviruses in different Aedes mosquito species in North Borneo Sabah, Malaysia |
| title_full | A standardized protocol for the detection of arboviruses in different Aedes mosquito species in North Borneo Sabah, Malaysia |
| title_fullStr | A standardized protocol for the detection of arboviruses in different Aedes mosquito species in North Borneo Sabah, Malaysia |
| title_full_unstemmed | A standardized protocol for the detection of arboviruses in different Aedes mosquito species in North Borneo Sabah, Malaysia |
| title_short | A standardized protocol for the detection of arboviruses in different Aedes mosquito species in North Borneo Sabah, Malaysia |
| title_sort | standardized protocol for the detection of arboviruses in different aedes mosquito species in north borneo sabah malaysia |
| topic | RNA extraction Aedes mosquitoes Multiplex real-time PCR Disease surveillance Dengue Zika |
| url | http://www.sciencedirect.com/science/article/pii/S2215016125003930 |
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