title: [Quantitative detection and species identificaton of human Plasmodium spp. by using SYBR Green I based real-time PCR].
abstract: OBJECTIVE
  To develop a real-time PCR method for human Plasmodium spp. qantitative detection and species identificaton.
  
  
  METHODS
  According to the sequence of Plasmodium 18S rRNA, the primer set was designed based on the genus-specific region around the species-specfic region. The PCR products were amplified and cloned into pGEM-T vector to produce standard plasmids of real-time PCR, and melting curve analysis was conducted following real-time PCR for Plasmodium species indentification.
  
  
  RESULTS
  By using the primer set, specific PCR products were produced from all of 4 human malaria parasites. The correlation of real-time PCR standard curve was good enough (r = -1.00) for quantitation. According to the melting curve analysis, the melting temperatures (Tm) of Plasmodium malariae, falciparum, ovale and vivax were significantly different, being 71.3, 72.8, 74.6 degrees C and 75.8 degrees C, respectively.
  
  
  CONCLUSIONS
  The SYBR Green I based real-time PCR method developed in this study can be used for human Plasmodium spp. quantitative detection and species identificaton.
authors.affiliation: Wuxi Entry-exit Inspection and Quarantine Bureau of PRC, Wuxi 214000, China.
authors.forename: Sheng-Qiang
authors.initials: SQ
authors.lastname: Wang
authors.forename: Hua-Yun
authors.initials: HY
authors.lastname: Zhou
authors.forename: Zong
authors.initials: Z
authors.lastname: Li
authors.forename: Yao-Bao
authors.initials: YB
authors.lastname: Liu
authors.forename: Xu-Feng
authors.initials: XF
authors.lastname: Fu
authors.forename: Jing-Jing
authors.initials: JJ
authors.lastname: Zhu
authors.forename: Jun
authors.initials: J
authors.lastname: Cao
authors.forename: Qi
authors.initials: Q
authors.lastname: Gao
mesh_terms: D001483:Base Sequence; D052160:Benzothiazoles; D016054:DNA, Protozoan / Q000235:genetics; D003959:Diamines; D006801:Humans; D008969:Molecular Sequence Data; D009930:Organic Chemicals / Q000737:chemistry; D010961:Plasmodium / Q000235:genetics / Q000302:isolation & purification*; D010965:Plasmodium malariae / Q000469:parasitology*; D011804:Quinolines; D060888:Real-Time Polymerase Chain Reaction / Q000295:instrumentation / Q000379:methods*
chemical_list: D052160:Benzothiazoles; D016054:DNA, Protozoan; D003959:Diamines; D009930:Organic Chemicals; D011804:Quinolines; C098022:SYBR Green I