title: [Establishment and evaluation of loop-mediated isothermal amplification based on Plasmodium falciparum unique genes coding PHIST proteins].
abstract: OBJECTIVE
  To establish a novel convenient loop-mediated isothermal amplification (LAMP) method with the unique genes coding Plasmodium helical interspersed sub-telomeric superfamily (PHIST) for the rapid molecular diagnosis of P. falciparum.
  
  
  METHODS
  The unique genes coding PHIST with high expression mRNA profile during the ring form or schizont period of P. falciparum were screened and selected from the PlasmoDB database. The LAMP primers of targeted genes were designed by the online software (PrimerExplorer V4). The LAMP assay was executed by the color-displaying method with SYBR Green. The dried blood spots of P. falciparum from clinical isolates were collected and the genomic DNA (gDNA) was extracted. For evaluation of sensitivity, the gDNA was diluted to four gradients (10⁻¹, 10⁻², 10⁻³, and 10⁻⁴). For assessment of specificity, the gDNA (s) of P. vivax, P. yoelii, Taenia saginata, and Schistosoma japonicum were also extracted.
  
  
  RESULTS
  Totally, 61 P. falciparum unique genes coding PHIST were found. The PF3D7_1372300 with high expression value during the ring form and PF3D7_1401600 with high expression value during the schizont period were selected for LAMP assay. The lowest detectable limits of PF3D7_1372300 and PF3D7_1401600 were 130.5 parasite/µl and 1305.3 parasite/µL, respectively. Specific tests showed the amplified products of P. falciparum was positive and all the others including P. vivax, P. yoelii, T. saginata, and S. japonicum were negative.
  
  
  CONCLUSIONS
  The established LAMP method with PF3D7_1372300 gene is sensitive, specific, simple and useful. It can be applied to the field investigation and clinical diagnosis for falciparum malaria.
authors.forename: Yi-jing
authors.initials: YJ
authors.lastname: Zhang
authors.forename: Bin
authors.initials: B
authors.lastname: Sun
authors.forename: Hua-fei
authors.initials: HF
authors.lastname: Shen
authors.forename: Kai
authors.initials: K
authors.lastname: Wu
authors.forename: Li-jun
authors.initials: LJ
authors.lastname: Song
authors.forename: Shuang
authors.initials: S
authors.lastname: Shen
authors.forename: Kai
authors.initials: K
authors.lastname: Li
authors.forename: Wen-yue
authors.initials: WY
authors.lastname: Xu
authors.forename: Yang
authors.initials: Y
authors.lastname: Dai
authors.forename: Min
authors.initials: M
authors.lastname: Lin
authors.forename: Shan
authors.initials: S
authors.lastname: Li
authors.forename: Wan-jun
authors.initials: WJ
authors.lastname: Wu
authors.forename: E-ping
authors.initials: EP
authors.lastname: Guo
authors.forename: Bei
authors.initials: B
authors.lastname: Li
authors.forename: Jian
authors.initials: J
authors.lastname: Li
mesh_terms: D001483:Base Sequence; D019295:Computational Biology; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D010963:Plasmodium falciparum / Q000235:genetics*; D015800:Protozoan Proteins / Q000235:genetics*
chemical_list: D015800:Protozoan Proteins