title: Enhanced Sensitivity for Detection of Plasmodium falciparum gametocytes by magnetic nanoparticles combined with enzyme substrate system.
abstract: The highly sensitive and specific detection of Pfg377 gene of Plasmodium falciparum gametocyte using Magnetic Nanoparticles PCR Enzyme-Linked Gene Assay (MELGA) was successfully developed. The MELGA included amplification of the Pfg377 gene by polymerase chain reaction (PCR) using magnetic nanoparticles (MNPs)-conjugated forward primer and biotinylated reverse primer, followed by post-analytical process using horseradish peroxidase (HRP)-conjugated streptavidin (SA). The complexes of MELGA product were incubated with the peroxidase substrate and hydrogen peroxide to produce the signal for colorimetric measurement. Altogether, the MELGA technique provided a highly sensitive and specific detection at 1 P. falciparum gametocyte/µL, which was more efficient than that of microscopic examination and rapid diagnostic tests (RDTs). Additionally, the MELGA could detect target gene at femtogram level, which was greater sensitive than the conventional PCR, nested PCR and loop-mediated isothermal amplification (LAMP). The MELGA technique could become a novel and practical method that overcome limitation of traditional gametocyte detection.
authors.affiliation: Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand.
authors.forename: Tienrat
authors.initials: T
authors.lastname: Tangchaikeeree
authors.affiliation: Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand.
authors.forename: Piamsiri
authors.initials: P
authors.lastname: Sawaisorn
authors.affiliation: Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Thammasat University, Pathum Thani 12120, Thailand.
authors.forename: Sangdao
authors.initials: S
authors.lastname: Somsri
authors.affiliation: National Nanotechnology Center, National Science and Technology Development Agency (NSTDA), Thailand Science Park, Pathum Thani 12120, Thailand.
authors.forename: Duangporn
authors.initials: D
authors.lastname: Polpanich
authors.affiliation: Molecular Biology of Malaria and Opportunistic Parasites Research Unit, Department of Parasitology, Faculty of Medicine, Chulalongkorn University, Bangkok 10330, Thailand.
authors.forename: Chaturong
authors.initials: C
authors.lastname: Putaporntip
authors.affiliation: Department of Chemistry, Faculty of Science, Mahidol University, Bangkok 10400, Thailand.
authors.forename: Pramuan
authors.initials: P
authors.lastname: Tangboriboonrat
authors.affiliation: Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand.
authors.forename: Rachanee
authors.initials: R
authors.lastname: Udomsangpetch
authors.affiliation: Center for Research and Innovation, Faculty of Medical Technology, Mahidol University, Bangkok 10700, Thailand. Electronic address: kulachart.jan@mahidol.ac.th.
authors.forename: Kulachart
authors.initials: K
authors.lastname: Jangpatarapongsa
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D017125:Genes, Protozoan / Q000235:genetics; D006735:Horseradish Peroxidase / Q000378:metabolism*; D057230:Limit of Detection; D059346:Magnets / Q000737:chemistry*; D053758:Nanoparticles / Q000737:chemistry*; D010963:Plasmodium falciparum / Q000166:cytology* / Q000235:genetics*
chemical_list: D017931:DNA Primers; D006735:Horseradish Peroxidase
keywords: Colorimetric system; Magnetic nanoparticles; Malaria gametocyte; Molecular detection; Plasmodium falciparum