title: Development and performance evaluation of a novel immunofluorescence chromatographic assay for histidine-rich protein 2 of Plasmodium falciparum.
abstract: BACKGROUND
  The low sensitivity and specificity of Plasmodium falciparum diagnostic tests pose a serious health threat to people living in endemic areas. The objective of the study was to develop a rapid assay for the detection of histidine-rich protein 2 (HRP2) of P. falciparum in whole blood by immunofluorescence chromatographic technology.
  
  
  METHODS
  A total of 1163 positive and negative blood samples were screened. The double-antibody sandwich assay was used to establish the kit and its performance was evaluated for sensitivity, specificity, accuracy, precision, stability, and clinical effectiveness.
  
  
  RESULTS
  The cut-off level of detection of the kit was 25 parasites/μl. Common interfering substances in human blood specimens, such as bilirubin, triglyceride and cholesterol had no significant effect on HRP2 antigen detection. The precision of the kit was run with different concentration of standard calibrators and the values were less than 10 %. The performance of this diagnostic kit in the detection of the calibrators has shown that a shelf life of about 12 months gives a more reliable result. Among clinical samples tested, the HRP2 test kit and the reference products had good coincidence rate in a parallel experiment and this test kit had a more sensitive detecting level to the target protein than the reference kits used in this study. The specificity and sensitivity for this test were 99.6 % (800/803) and 99.7 % (1160/1163), respectively.
  
  
  CONCLUSIONS
  A novel HRP2 immunofluorescence detection method was developed in this study. Overall performance evaluation indicated that the kit has a rapid, high sensitivity and on-spot method for detecting P. falciparum.
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006, Guangdong, China. keren_kang@hotmail.com.|National Engineering Laboratory of Rapid Diagnostic Tests, Guangzhou Wondfo Biotech Co Ltd, Guangzhou, 510663, Guangdong, China. keren_kang@hotmail.com.
authors.forename: Keren
authors.initials: K
authors.lastname: Kang
authors.affiliation: Department of Molecular Biology and Biotechnology, School of Biological Sciences, College of Agriculture and Natural Sciences, University of Cape Coast, Cape Coast, Ghana. deek2083@yahoo.co.uk.
authors.forename: Emmanuel E
authors.initials: EE
authors.lastname: Dzakah
authors.affiliation: National Engineering Laboratory of Rapid Diagnostic Tests, Guangzhou Wondfo Biotech Co Ltd, Guangzhou, 510663, Guangdong, China. 1060876084@qq.com.
authors.forename: Yongping
authors.initials: Y
authors.lastname: Huang
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006, Guangdong, China. xiemq@gdstc.gov.cn.
authors.forename: Mingquan
authors.initials: M
authors.lastname: Xie
authors.affiliation: School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006, Guangdong, China. xcluo@scut.edu.cn.
authors.forename: Xiaochun
authors.initials: X
authors.lastname: Luo
authors.affiliation: National Engineering Laboratory of Rapid Diagnostic Tests, Guangzhou Wondfo Biotech Co Ltd, Guangzhou, 510663, Guangdong, China. lwm@wondfo.com.cn.
authors.forename: Wenmei
authors.initials: W
authors.lastname: Li
authors.affiliation: National Engineering Laboratory of Rapid Diagnostic Tests, Guangzhou Wondfo Biotech Co Ltd, Guangzhou, 510663, Guangdong, China. wjh@wondfo.com.cn.
authors.forename: Jihua
authors.initials: J
authors.lastname: Wang
mesh_terms: D000953:Antigens, Protozoan / Q000302:isolation & purification*; D002845:Chromatography / Q000379:methods*; D003955:Diagnostic Tests, Routine / Q000379:methods*; D005455:Fluorescent Antibody Technique; D016778:Malaria, Falciparum / Q000175:diagnosis*; D010963:Plasmodium falciparum / Q000502:physiology*; D015800:Protozoan Proteins / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
chemical_list: D000953:Antigens, Protozoan; C514457:HRP-2 antigen, Plasmodium falciparum; D015800:Protozoan Proteins