Point-of-Care Diagnosis of Malaria Using a Simple, Purification-Free DNA Extraction Method Coupled with Loop-Mediated Isothermal Amplification-Lateral Flow
We propose a protocol suitable for point-of-care diagnosis of malaria utilizing a simple and purification-free DNA extraction method with the combination of loop-mediated isothermal amplification assay and lateral flow (LAMP-LF). The multiplex LAMP-LF platform developed here can simultaneously detec...
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| Format: | Article |
| Language: | English |
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MDPI AG
2023-03-01
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| Series: | Tropical Medicine and Infectious Disease |
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| Online Access: | https://www.mdpi.com/2414-6366/8/4/199 |
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| author | Meng Yee Lai Lee Phone Youth Zen Mohd Hafizi Abdul Hamid Jenarun Jelip Rose Nani Mudin Vun Jan Shui Ivan Lee Ngie Ping Francis Izreena Saihidi Yee Ling Lau |
| author_facet | Meng Yee Lai Lee Phone Youth Zen Mohd Hafizi Abdul Hamid Jenarun Jelip Rose Nani Mudin Vun Jan Shui Ivan Lee Ngie Ping Francis Izreena Saihidi Yee Ling Lau |
| author_sort | Meng Yee Lai |
| collection | DOAJ |
| description | We propose a protocol suitable for point-of-care diagnosis of malaria utilizing a simple and purification-free DNA extraction method with the combination of loop-mediated isothermal amplification assay and lateral flow (LAMP-LF). The multiplex LAMP-LF platform developed here can simultaneously detect <i>Plasmodium knowlesi, P. vivax, P. falciparum,</i> and <i>Plasmodium</i> genus (for <i>P. malariae</i> and <i>P. ovale</i>). Through the capillary effect, the results can be observed by the red band signal on the test and control lines within 5 min. The developed multiplex LAMP-LF was tested with 86 clinical blood samples on-site at Hospital Kapit, Sarawak, Malaysia. By using microscopy as the reference method, the multiplex LAMP-LF showed 100% sensitivity (95% confidence interval (CI): 91.4 to 100.00%) and 97.8% specificity (95% CI: 88.2% to 99.9%). The high sensitivity and specificity of multiplex LAMP-LF make it ideal for use as a point-of-care diagnostic tool. The simple and purification-free DNA extraction protocol can be employed as an alternative DNA extraction method for malaria diagnosis in resource-limited settings. By combining the simple DNA extraction protocol and multiplex LAMP-LF approach, we aim to develop a simple-to-handle and easy-to-read molecular diagnostic tool for malaria in both laboratory and on-site settings. |
| first_indexed | 2024-03-11T04:28:20Z |
| format | Article |
| id | doaj.art-aaa664de86f24d9698932147fd0701f0 |
| institution | Directory Open Access Journal |
| issn | 2414-6366 |
| language | English |
| last_indexed | 2024-03-11T04:28:20Z |
| publishDate | 2023-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Tropical Medicine and Infectious Disease |
| spelling | doaj.art-aaa664de86f24d9698932147fd0701f02023-11-17T21:39:30ZengMDPI AGTropical Medicine and Infectious Disease2414-63662023-03-018419910.3390/tropicalmed8040199Point-of-Care Diagnosis of Malaria Using a Simple, Purification-Free DNA Extraction Method Coupled with Loop-Mediated Isothermal Amplification-Lateral FlowMeng Yee Lai0Lee Phone Youth Zen1Mohd Hafizi Abdul Hamid2Jenarun Jelip3Rose Nani Mudin4Vun Jan Shui Ivan5Lee Ngie Ping Francis6Izreena Saihidi7Yee Ling Lau8Department of Parasitology, Faculty of Medicine, Universiti Malaya, Kuala Lumpur 50603, MalaysiaDepartment of Parasitology, Faculty of Medicine, Universiti Malaya, Kuala Lumpur 50603, MalaysiaVector Borne Disease Sector, Ministry of Health, Putrajaya 62590, MalaysiaVector Borne Disease Sector, Ministry of Health, Putrajaya 62590, MalaysiaVector Borne Disease Sector, Ministry of Health, Putrajaya 62590, MalaysiaHospital Kapit, Pathology Unit, Jalan Mamora, Kapit 96800, MalaysiaHospital Kapit, Pathology Unit, Jalan Mamora, Kapit 96800, MalaysiaHospital Kapit, Pathology Unit, Jalan Mamora, Kapit 96800, MalaysiaDepartment of Parasitology, Faculty of Medicine, Universiti Malaya, Kuala Lumpur 50603, MalaysiaWe propose a protocol suitable for point-of-care diagnosis of malaria utilizing a simple and purification-free DNA extraction method with the combination of loop-mediated isothermal amplification assay and lateral flow (LAMP-LF). The multiplex LAMP-LF platform developed here can simultaneously detect <i>Plasmodium knowlesi, P. vivax, P. falciparum,</i> and <i>Plasmodium</i> genus (for <i>P. malariae</i> and <i>P. ovale</i>). Through the capillary effect, the results can be observed by the red band signal on the test and control lines within 5 min. The developed multiplex LAMP-LF was tested with 86 clinical blood samples on-site at Hospital Kapit, Sarawak, Malaysia. By using microscopy as the reference method, the multiplex LAMP-LF showed 100% sensitivity (95% confidence interval (CI): 91.4 to 100.00%) and 97.8% specificity (95% CI: 88.2% to 99.9%). The high sensitivity and specificity of multiplex LAMP-LF make it ideal for use as a point-of-care diagnostic tool. The simple and purification-free DNA extraction protocol can be employed as an alternative DNA extraction method for malaria diagnosis in resource-limited settings. By combining the simple DNA extraction protocol and multiplex LAMP-LF approach, we aim to develop a simple-to-handle and easy-to-read molecular diagnostic tool for malaria in both laboratory and on-site settings.https://www.mdpi.com/2414-6366/8/4/199malariaDNA extractionLAMPmolecular diagnosisfilter paper-basedlateral flow |
| spellingShingle | Meng Yee Lai Lee Phone Youth Zen Mohd Hafizi Abdul Hamid Jenarun Jelip Rose Nani Mudin Vun Jan Shui Ivan Lee Ngie Ping Francis Izreena Saihidi Yee Ling Lau Point-of-Care Diagnosis of Malaria Using a Simple, Purification-Free DNA Extraction Method Coupled with Loop-Mediated Isothermal Amplification-Lateral Flow Tropical Medicine and Infectious Disease malaria DNA extraction LAMP molecular diagnosis filter paper-based lateral flow |
| title | Point-of-Care Diagnosis of Malaria Using a Simple, Purification-Free DNA Extraction Method Coupled with Loop-Mediated Isothermal Amplification-Lateral Flow |
| title_full | Point-of-Care Diagnosis of Malaria Using a Simple, Purification-Free DNA Extraction Method Coupled with Loop-Mediated Isothermal Amplification-Lateral Flow |
| title_fullStr | Point-of-Care Diagnosis of Malaria Using a Simple, Purification-Free DNA Extraction Method Coupled with Loop-Mediated Isothermal Amplification-Lateral Flow |
| title_full_unstemmed | Point-of-Care Diagnosis of Malaria Using a Simple, Purification-Free DNA Extraction Method Coupled with Loop-Mediated Isothermal Amplification-Lateral Flow |
| title_short | Point-of-Care Diagnosis of Malaria Using a Simple, Purification-Free DNA Extraction Method Coupled with Loop-Mediated Isothermal Amplification-Lateral Flow |
| title_sort | point of care diagnosis of malaria using a simple purification free dna extraction method coupled with loop mediated isothermal amplification lateral flow |
| topic | malaria DNA extraction LAMP molecular diagnosis filter paper-based lateral flow |
| url | https://www.mdpi.com/2414-6366/8/4/199 |
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