Validation of <i>Pf</i>SNP-LAMP-Lateral Flow Dipstick for Detection of Single Nucleotide Polymorphism Associated with Pyrimethamine Resistance in <i>Plasmodium falciparum</i>
The loop-mediated isothermal amplification coupled with lateral flow dipstick (<i>Pf</i>SNP-LAMP-LFD) was recently developed to detect single nucleotide polymorphism (A<b>A</b>T → A<b>T</b>T), corresponding to substitution of asparagine to isoleucine at amino acid...
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MDPI AG
2020-11-01
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| Series: | Diagnostics |
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| Online Access: | https://www.mdpi.com/2075-4418/10/11/948 |
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| author | Suganya Yongkiettrakul Fassou René Kolié Darin Kongkasuriyachai Jetsumon Sattabongkot Wang Nguitragool Namfon Nawattanapaibool Chayanut Suansomjit Saradee Warit Niwat Kangwanrangsan Sureemas Buates |
| author_facet | Suganya Yongkiettrakul Fassou René Kolié Darin Kongkasuriyachai Jetsumon Sattabongkot Wang Nguitragool Namfon Nawattanapaibool Chayanut Suansomjit Saradee Warit Niwat Kangwanrangsan Sureemas Buates |
| author_sort | Suganya Yongkiettrakul |
| collection | DOAJ |
| description | The loop-mediated isothermal amplification coupled with lateral flow dipstick (<i>Pf</i>SNP-LAMP-LFD) was recently developed to detect single nucleotide polymorphism (A<b>A</b>T → A<b>T</b>T), corresponding to substitution of asparagine to isoleucine at amino acid position 51 in the <i>P. falciparum</i><i>dhfr-ts</i> gene associated with antifolate resistance. In this present study, the <i>Pf</i>SNP-LAMP-LFD was validated on 128 clinical malaria samples of broad ranged parasite densities (10 to 87,634 parasites per microliter of blood). The results showed 100% accuracy for the detection of single nucleotide polymorphism for N51I mutation. Indeed, the high prevalence of N51I in the <i>Pfdhfr-ts</i> gene detected in the clinical samples is in line with reports of widespread antifolate resistant <i>P. falciparum</i> in Thailand. The relationship between enzyme choice and reaction time was observed to have an effect on <i>Pf</i>SNP-LAMP-LFD specificity; however, the method yielded consistent results once the conditions have been optimized. The results demonstrate that <i>Pf</i>SNP-LAMP-LFD is a simple method with sufficient sensitivity and specificity to be deployed in routine surveillance of antifolate resistance molecular marker and inform antimalarial management policy. |
| first_indexed | 2024-03-10T14:51:37Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2075-4418 |
| language | English |
| last_indexed | 2024-03-10T14:51:37Z |
| publishDate | 2020-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Diagnostics |
| spelling | doaj.art-37b9b7173c7a40aa841fe6c0bf3e82602023-11-20T20:53:43ZengMDPI AGDiagnostics2075-44182020-11-01101194810.3390/diagnostics10110948Validation of <i>Pf</i>SNP-LAMP-Lateral Flow Dipstick for Detection of Single Nucleotide Polymorphism Associated with Pyrimethamine Resistance in <i>Plasmodium falciparum</i>Suganya Yongkiettrakul0Fassou René Kolié1Darin Kongkasuriyachai2Jetsumon Sattabongkot3Wang Nguitragool4Namfon Nawattanapaibool5Chayanut Suansomjit6Saradee Warit7Niwat Kangwanrangsan8Sureemas Buates9National Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, ThailandDepartment of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, ThailandNational Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, ThailandMahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, ThailandDepartment of Molecular Tropical Medicine and Genetics, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, ThailandDepartment of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, ThailandMahidol Vivax Research Unit, Faculty of Tropical Medicine, Mahidol University, Bangkok 10400, ThailandNational Center for Genetic Engineering and Biotechnology (BIOTEC), National Science and Technology Development Agency (NSTDA), Pathum Thani 12120, ThailandDepartment of Pathobiology, Faculty of Science, Mahidol University, Bangkok 10400, ThailandDepartment of Microbiology, Faculty of Science, Mahidol University, Bangkok 10400, ThailandThe loop-mediated isothermal amplification coupled with lateral flow dipstick (<i>Pf</i>SNP-LAMP-LFD) was recently developed to detect single nucleotide polymorphism (A<b>A</b>T → A<b>T</b>T), corresponding to substitution of asparagine to isoleucine at amino acid position 51 in the <i>P. falciparum</i><i>dhfr-ts</i> gene associated with antifolate resistance. In this present study, the <i>Pf</i>SNP-LAMP-LFD was validated on 128 clinical malaria samples of broad ranged parasite densities (10 to 87,634 parasites per microliter of blood). The results showed 100% accuracy for the detection of single nucleotide polymorphism for N51I mutation. Indeed, the high prevalence of N51I in the <i>Pfdhfr-ts</i> gene detected in the clinical samples is in line with reports of widespread antifolate resistant <i>P. falciparum</i> in Thailand. The relationship between enzyme choice and reaction time was observed to have an effect on <i>Pf</i>SNP-LAMP-LFD specificity; however, the method yielded consistent results once the conditions have been optimized. The results demonstrate that <i>Pf</i>SNP-LAMP-LFD is a simple method with sufficient sensitivity and specificity to be deployed in routine surveillance of antifolate resistance molecular marker and inform antimalarial management policy.https://www.mdpi.com/2075-4418/10/11/948dihydrofolate reductaseloop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD)malaria detection<i>Plasmodium falciparum</i>pyrimethamineantifolate resistance |
| spellingShingle | Suganya Yongkiettrakul Fassou René Kolié Darin Kongkasuriyachai Jetsumon Sattabongkot Wang Nguitragool Namfon Nawattanapaibool Chayanut Suansomjit Saradee Warit Niwat Kangwanrangsan Sureemas Buates Validation of <i>Pf</i>SNP-LAMP-Lateral Flow Dipstick for Detection of Single Nucleotide Polymorphism Associated with Pyrimethamine Resistance in <i>Plasmodium falciparum</i> Diagnostics dihydrofolate reductase loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) malaria detection <i>Plasmodium falciparum</i> pyrimethamine antifolate resistance |
| title | Validation of <i>Pf</i>SNP-LAMP-Lateral Flow Dipstick for Detection of Single Nucleotide Polymorphism Associated with Pyrimethamine Resistance in <i>Plasmodium falciparum</i> |
| title_full | Validation of <i>Pf</i>SNP-LAMP-Lateral Flow Dipstick for Detection of Single Nucleotide Polymorphism Associated with Pyrimethamine Resistance in <i>Plasmodium falciparum</i> |
| title_fullStr | Validation of <i>Pf</i>SNP-LAMP-Lateral Flow Dipstick for Detection of Single Nucleotide Polymorphism Associated with Pyrimethamine Resistance in <i>Plasmodium falciparum</i> |
| title_full_unstemmed | Validation of <i>Pf</i>SNP-LAMP-Lateral Flow Dipstick for Detection of Single Nucleotide Polymorphism Associated with Pyrimethamine Resistance in <i>Plasmodium falciparum</i> |
| title_short | Validation of <i>Pf</i>SNP-LAMP-Lateral Flow Dipstick for Detection of Single Nucleotide Polymorphism Associated with Pyrimethamine Resistance in <i>Plasmodium falciparum</i> |
| title_sort | validation of i pf i snp lamp lateral flow dipstick for detection of single nucleotide polymorphism associated with pyrimethamine resistance in i plasmodium falciparum i |
| topic | dihydrofolate reductase loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) malaria detection <i>Plasmodium falciparum</i> pyrimethamine antifolate resistance |
| url | https://www.mdpi.com/2075-4418/10/11/948 |
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