Multiplex PCR Assay for the Identification of Four Species of the <i>Anopheles</i> Leucosphyrus Sub-Group in Malaysia
The Leucosphyrus Group of mosquitoes are the major simian malaria vectors in Malaysia. Accurate species identification is required to help in curbing the spread of simian malaria. The aim of the study is to provide an accurate molecular method for identifying the four important <i>Anopheles<...
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MDPI AG
2022-02-01
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| Series: | Insects |
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| Online Access: | https://www.mdpi.com/2075-4450/13/2/195 |
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| author | Sandthya Pramasivan Jonathan Wee Kent Liew Nantha Kumar Jeyaprakasam Van Lun Low Romano Ngui Indra Vythilingam |
| author_facet | Sandthya Pramasivan Jonathan Wee Kent Liew Nantha Kumar Jeyaprakasam Van Lun Low Romano Ngui Indra Vythilingam |
| author_sort | Sandthya Pramasivan |
| collection | DOAJ |
| description | The Leucosphyrus Group of mosquitoes are the major simian malaria vectors in Malaysia. Accurate species identification is required to help in curbing the spread of simian malaria. The aim of the study is to provide an accurate molecular method for identifying the four important <i>Anopheles</i> vector species found in Malaysia. Mosquito specimens were collected from various localities in Malaysia, where simian malaria cases were reported. DNA from 122 mosquito specimens was tested to develop a multiplex polymerase chain reaction (PCR) assay. The specificity of this assay was tested against other mosquito species. Molecular identification of the species was further confirmed by analysing the internal transcribed spacer 2 (<i>ITS2</i>) DNA region of the specimens. <i>Anopheles balabacensis</i> and <i>An. latens</i> showed two distinct clades in the phylogenetic tree. The multiplex PCR assay was developed based on the <i>ITS2</i> region for the identification of <i>Anopheles introlatus</i> (298–299 bp), <i>Anopheles latens</i> (197–198 bp), <i>Anopheles cracens</i> (421–426 bp), and <i>Anopheles balabacensis</i> (224–228 bp). This method will be useful to accurately identify the major <i>Anopheles</i> Leucosphyrus Group species in Malaysia, which are difficult to identify morphologically, to determine the correct vector as well as its geographical distribution. |
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| language | English |
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| spelling | doaj.art-1fded42245a547ab9fa758e7fec03d252023-11-23T20:27:42ZengMDPI AGInsects2075-44502022-02-0113219510.3390/insects13020195Multiplex PCR Assay for the Identification of Four Species of the <i>Anopheles</i> Leucosphyrus Sub-Group in MalaysiaSandthya Pramasivan0Jonathan Wee Kent Liew1Nantha Kumar Jeyaprakasam2Van Lun Low3Romano Ngui4Indra Vythilingam5Department of Parasitology, Faculty of Medicine, University Malaya, Kuala Lumpur 50603, MalaysiaDepartment of Parasitology, Faculty of Medicine, University Malaya, Kuala Lumpur 50603, MalaysiaDepartment of Parasitology, Faculty of Medicine, University Malaya, Kuala Lumpur 50603, MalaysiaTropical Infectious Diseases Research & Education Centre (TIDREC), University Malaya, Kuala Lumpur 50603, MalaysiaDepartment of Parasitology, Faculty of Medicine, University Malaya, Kuala Lumpur 50603, MalaysiaDepartment of Parasitology, Faculty of Medicine, University Malaya, Kuala Lumpur 50603, MalaysiaThe Leucosphyrus Group of mosquitoes are the major simian malaria vectors in Malaysia. Accurate species identification is required to help in curbing the spread of simian malaria. The aim of the study is to provide an accurate molecular method for identifying the four important <i>Anopheles</i> vector species found in Malaysia. Mosquito specimens were collected from various localities in Malaysia, where simian malaria cases were reported. DNA from 122 mosquito specimens was tested to develop a multiplex polymerase chain reaction (PCR) assay. The specificity of this assay was tested against other mosquito species. Molecular identification of the species was further confirmed by analysing the internal transcribed spacer 2 (<i>ITS2</i>) DNA region of the specimens. <i>Anopheles balabacensis</i> and <i>An. latens</i> showed two distinct clades in the phylogenetic tree. The multiplex PCR assay was developed based on the <i>ITS2</i> region for the identification of <i>Anopheles introlatus</i> (298–299 bp), <i>Anopheles latens</i> (197–198 bp), <i>Anopheles cracens</i> (421–426 bp), and <i>Anopheles balabacensis</i> (224–228 bp). This method will be useful to accurately identify the major <i>Anopheles</i> Leucosphyrus Group species in Malaysia, which are difficult to identify morphologically, to determine the correct vector as well as its geographical distribution.https://www.mdpi.com/2075-4450/13/2/195species identificationmultiplex PCR assay<i>Anopheles</i>simian malaria<i>ITS2</i>Malaysia |
| spellingShingle | Sandthya Pramasivan Jonathan Wee Kent Liew Nantha Kumar Jeyaprakasam Van Lun Low Romano Ngui Indra Vythilingam Multiplex PCR Assay for the Identification of Four Species of the <i>Anopheles</i> Leucosphyrus Sub-Group in Malaysia Insects species identification multiplex PCR assay <i>Anopheles</i> simian malaria <i>ITS2</i> Malaysia |
| title | Multiplex PCR Assay for the Identification of Four Species of the <i>Anopheles</i> Leucosphyrus Sub-Group in Malaysia |
| title_full | Multiplex PCR Assay for the Identification of Four Species of the <i>Anopheles</i> Leucosphyrus Sub-Group in Malaysia |
| title_fullStr | Multiplex PCR Assay for the Identification of Four Species of the <i>Anopheles</i> Leucosphyrus Sub-Group in Malaysia |
| title_full_unstemmed | Multiplex PCR Assay for the Identification of Four Species of the <i>Anopheles</i> Leucosphyrus Sub-Group in Malaysia |
| title_short | Multiplex PCR Assay for the Identification of Four Species of the <i>Anopheles</i> Leucosphyrus Sub-Group in Malaysia |
| title_sort | multiplex pcr assay for the identification of four species of the i anopheles i leucosphyrus sub group in malaysia |
| topic | species identification multiplex PCR assay <i>Anopheles</i> simian malaria <i>ITS2</i> Malaysia |
| url | https://www.mdpi.com/2075-4450/13/2/195 |
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