WHO, 2017. The World Malaria Report 2017. Geneva, Switzerland: World Health Organization.
Singh B, 2016. Plasmodium knowlesi: an update. Microbiol Aust 37: 39–42.
Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T, 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28: E63.
Nagamine K, Hase T, Notomi T, 2002. Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes 16: 223–229.
Yongkiettrakul S, Jaroenram W, Arunrut N, Chareanchim W, Pannengpetch S, Suebsing R, Kiatpathomchai W, Pornthanakasem W, Yuthavong Y, Kongkasuriyachai D, 2014. Application of loop-mediated isothermal amplification assay combined with lateral flow dipstick for detection of Plasmodium falciparum and Plasmodium vivax. Parasitol Int 63: 777–784.
Lau YL, Lai MY, Fong MY, Jelip J, Mahmud R, 2016. Loop-mediated isothermal amplification assay for identification of five human Plasmodium species in Malaysia. Am J Trop Med Hyg 94: 336–339.
Han ET, Watanabe R, Sattabongkot J, Khuntirat B, Sirichaisinthop J, Iriko H, Jin L, Takeo S, Tsuboi T, 2007. Detection of four Plasmodium species by genus- and species-specific loop-mediated isothermal amplification for clinical diagnosis. J Clin Microbiol 45: 2521–2528.
Li Y, Kumar N, Gopalakrishnan A, Ginocchio C, Manji R, Bythrow M, Lemieux B, Kong H, 2013. Detection and species identification of malaria parasites by isothermal tHDA amplification directly from human blood without sample preparation. J Mol Diagn 15: 634–641.
Zhang Y, Yao Y, Du W, Wu K, Xu W, Lin M, Tan H, Li J, 2017. Development of loop-mediated isothermal amplification with Plasmodium falciparum unique genes for molecular diagnosis of human malaria. Pathog Glob Health 111: 247–255.
Imai K et al. 2017. A novel diagnostic method for malaria using loop-mediated isothermal amplification (LAMP) and MinION™ nanopore sequencer. BMC Infect Dis 17: 621.
Piera KA, Aziz A, William T, Bell D, González IJ, Barber BE, Anstey NM, Grigg MJ, 2017. Detection of Plasmodium knowlesi, Plasmodium falciparum and Plasmodium vivax using loop-mediated isothermal amplification (LAMP) in a co-endemic area in Malaysia. Malar J 16: 29.
Patel JC et al. 2013. Real-time loop-mediated isothermal amplification (RealAmp) for the species-specific identification of Plasmodium vivax. PLoS One 8: e54986.
Sattabongkot J, Tsuboi T, Han ET, Bantuchai S, Buates S, 2014. Loop-mediated isothermal amplification assay for rapid diagnosis of malaria infections in an area of endemicity in Thailand. J Clin Microbiol 52: 1471–1477.
Lalle M, Possenti A, Dubey JP, Pozio E, 2017. Loop-mediated isothermal amplification-lateral-flow dipstick (LAMP-LFD) to detect Toxoplasma gondii oocyst in ready-to-eat salad. Food Microbiol 70: 137–142.
Wang Y, Li H, Wang Y, Zhang L, Xu J, Ye C, 2017. Loop-mediated isothermal amplification label-based gold nanoparticles lateral flow biosensor for detection of Enterococcus faecalis and Staphylococcus aureus. Front Microbiol 8: 192.
Foo FP, Chan YY, Mohamed M, Wong WK, Najian N, Lim BH, 2017. Development of a thermostabilised triplex LAMP assay with dry-reagent four target lateral flow dipstick for detection of Entamoeba histolytica and non-pathogenic Entamoeba spp. Anal Chim Acta 966: 71–80.
Al-Soud WA, Radstrom P, 2001. Purification and characterization of PCR inhibitory components in blood cells. J Clin Microbiol 39: 485–493.
Guthrie R, Susi A, 1963. A simple phenylalanine method for detecting phenylketonuria in large populations of newborn infants. Pediatrics 32: 338–343.
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The present study aims to develop a method for rapid diagnosis of malaria using loop-mediated isothermal amplification (LAMP) combined with a lateral flow device (LFD). By adding the biotin-labeled and fluorescein amidite-labeled loop primers to the LAMP reaction solution, the end product can be visualized on a LFD. The entire procedure takes approximately 42 minutes to complete, LAMP assay exhibited high sensitivity, as the detection limit was 0.01 pg/μL for all five Plasmodium species. It was demonstrated that all Plasmodium knowlesi (N = 90) and Plasmodium vivax (N = 56) were positively amplified by LAMP-LFD assay, whereas healthy donor samples (N = 8) were negative. However, not all mixed infections were positive, and other infected nonmalaria samples were negative. Loop-mediated isothermal amplification-LFD represents a robust approach with potential suitability for use in resource-constrained laboratories. We believe that LAMP-LFD has a potential to be developed as point-of-care diagnostic tool in future.
These authors contributed equally to this work.
Financial support: This study was supported by University Malaya PPP Research Grant (PG066-2016A) and Department of Biotechnology under SIBRI Project on Malarial Diagnostics (File no: BT/SBIRI/358/57-B6/2007).
Authors’ addresses: Prudhvi Chand Mallepaddi and Rathnagiri Polavarapu, Genomix Molecular Diagnostics Pvt. Ltd., Hyderabad, India, E-mails: prudhvi.mallepaddi@genomixbiotech.com and giri@genomixbiotech.com. Meng-Yee Lai, Jonathan Wee-Kent Liew, and Yee-Ling Lau, Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia, E-mails: mengylai11@yahoo.com, jon_wkent@hotmail.com, and lauyeeling@um.edu.my. Sudhakar Podha, Department of Biotechnology, Acharya Nagarjuna University, Guntur, India, E-mail: sudhakarpodha@gmail.com. Choo-Huck Ooi, Sarawak State Health Department, Jalan Diplomatik, Kuching, Malaysia, E-mail: ooi.choo.huck@gmail.com.
WHO, 2017. The World Malaria Report 2017. Geneva, Switzerland: World Health Organization.
Singh B, 2016. Plasmodium knowlesi: an update. Microbiol Aust 37: 39–42.
Notomi T, Okayama H, Masubuchi H, Yonekawa T, Watanabe K, Amino N, Hase T, 2000. Loop-mediated isothermal amplification of DNA. Nucleic Acids Res 28: E63.
Nagamine K, Hase T, Notomi T, 2002. Accelerated reaction by loop-mediated isothermal amplification using loop primers. Mol Cell Probes 16: 223–229.
Yongkiettrakul S, Jaroenram W, Arunrut N, Chareanchim W, Pannengpetch S, Suebsing R, Kiatpathomchai W, Pornthanakasem W, Yuthavong Y, Kongkasuriyachai D, 2014. Application of loop-mediated isothermal amplification assay combined with lateral flow dipstick for detection of Plasmodium falciparum and Plasmodium vivax. Parasitol Int 63: 777–784.
Lau YL, Lai MY, Fong MY, Jelip J, Mahmud R, 2016. Loop-mediated isothermal amplification assay for identification of five human Plasmodium species in Malaysia. Am J Trop Med Hyg 94: 336–339.
Han ET, Watanabe R, Sattabongkot J, Khuntirat B, Sirichaisinthop J, Iriko H, Jin L, Takeo S, Tsuboi T, 2007. Detection of four Plasmodium species by genus- and species-specific loop-mediated isothermal amplification for clinical diagnosis. J Clin Microbiol 45: 2521–2528.
Li Y, Kumar N, Gopalakrishnan A, Ginocchio C, Manji R, Bythrow M, Lemieux B, Kong H, 2013. Detection and species identification of malaria parasites by isothermal tHDA amplification directly from human blood without sample preparation. J Mol Diagn 15: 634–641.
Zhang Y, Yao Y, Du W, Wu K, Xu W, Lin M, Tan H, Li J, 2017. Development of loop-mediated isothermal amplification with Plasmodium falciparum unique genes for molecular diagnosis of human malaria. Pathog Glob Health 111: 247–255.
Imai K et al. 2017. A novel diagnostic method for malaria using loop-mediated isothermal amplification (LAMP) and MinION™ nanopore sequencer. BMC Infect Dis 17: 621.
Piera KA, Aziz A, William T, Bell D, González IJ, Barber BE, Anstey NM, Grigg MJ, 2017. Detection of Plasmodium knowlesi, Plasmodium falciparum and Plasmodium vivax using loop-mediated isothermal amplification (LAMP) in a co-endemic area in Malaysia. Malar J 16: 29.
Patel JC et al. 2013. Real-time loop-mediated isothermal amplification (RealAmp) for the species-specific identification of Plasmodium vivax. PLoS One 8: e54986.
Sattabongkot J, Tsuboi T, Han ET, Bantuchai S, Buates S, 2014. Loop-mediated isothermal amplification assay for rapid diagnosis of malaria infections in an area of endemicity in Thailand. J Clin Microbiol 52: 1471–1477.
Lalle M, Possenti A, Dubey JP, Pozio E, 2017. Loop-mediated isothermal amplification-lateral-flow dipstick (LAMP-LFD) to detect Toxoplasma gondii oocyst in ready-to-eat salad. Food Microbiol 70: 137–142.
Wang Y, Li H, Wang Y, Zhang L, Xu J, Ye C, 2017. Loop-mediated isothermal amplification label-based gold nanoparticles lateral flow biosensor for detection of Enterococcus faecalis and Staphylococcus aureus. Front Microbiol 8: 192.
Foo FP, Chan YY, Mohamed M, Wong WK, Najian N, Lim BH, 2017. Development of a thermostabilised triplex LAMP assay with dry-reagent four target lateral flow dipstick for detection of Entamoeba histolytica and non-pathogenic Entamoeba spp. Anal Chim Acta 966: 71–80.
Al-Soud WA, Radstrom P, 2001. Purification and characterization of PCR inhibitory components in blood cells. J Clin Microbiol 39: 485–493.
Guthrie R, Susi A, 1963. A simple phenylalanine method for detecting phenylketonuria in large populations of newborn infants. Pediatrics 32: 338–343.
Past two years | Past Year | Past 30 Days | |
---|---|---|---|
Abstract Views | 2692 | 2429 | 1531 |
Full Text Views | 949 | 23 | 15 |
PDF Downloads | 407 | 14 | 5 |