title: LAMP kit for diagnosis of non-falciparum malaria in Plasmodium ovale infected patients.
abstract: Microscopy and rapid diagnosis tests have a limited sensitivity in diagnosis of malaria by Plasmodium ovale. The LAMP kit (LoopAMP®) can be used in the field without special equipment and could have an important role in malaria control programmes in endemic areas and for malaria diagnosis in returned travellers. The performance of the Pan primer of the kit in detecting malaria by P. ovale was compared with the results of standard nPCR in samples of patients returning from P. ovale endemic areas.
  
  
  
  Plasmodium ovale positive samples (29, tested by PCR and/or microscopy) and malaria negative specimens (398, tested by microscopy and PCR) were collected in different hospitals of Europe from June 2014 to March 2016 and frozen at -20 °C. Boil and spin method was used to extract DNA from all samples and amplification was performed with LoopAMP® MALARIA kit (Eiken Chemical, Japan) in an automated turbidimeter (Eiken 500). The results of LAMP read by turbidimetry and with the naked eye were compared.
  
  
  
  The kit showed a sensitivity of 100% and a specificity of 97.24% with positive and negative predictive values of 72.5 and 100%, respectively. Naked eyed readings were in accordance with turbidimetry readings (sensitivity, 92.5%, specificity, 98.96% and positive and negative predictive values, respectively, 90.24 and 99.22%). The limit of detection of LAMP assay for P. ovale was between 0.8 and 2 parasites/µl.
  
  
  
  The Pan primer of the Malaria kit LoopAMP® can detect P. ovale at very low-levels and showed a predictive negative value of 100%. This tool can be useful in malaria control and elimination programmes and in returned travellers from P. ovale endemic areas. Naked eye readings are equivalent to automated turbidimeter readings in specimens obtained with EDTA.
authors.affiliation: Department of Clinical Microbiology and Parasitology, Hospital Príncipe de Asturias, 28805, Alcalá de Henares, Madrid, Spain. jcuadros48@gmail.com.
authors.forename: Juan
authors.identifier: 0000-0003-4098-4377
authors.initials: J
authors.lastname: Cuadros
authors.affiliation: Department of Clinical Microbiology and Parasitology, Hospital de la Princesa, Madrid, Spain.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Martin Ramírez
authors.affiliation: Foundation for Innovative New Diagnostics (FIND), Geneva, Switzerland.
authors.forename: Iveth J
authors.initials: IJ
authors.lastname: González
authors.affiliation: Foundation for Innovative New Diagnostics (FIND), Geneva, Switzerland.
authors.forename: Xavier C
authors.initials: XC
authors.lastname: Ding
authors.affiliation: Department of Medicine and Laboratory, Gambo Rural General Hospital, Kore, West-Arsi, Gambo, Ethiopia.|Department of Otorhinolaryngology-Head and Neck Surgery, University of Helsinki and Helsinki University Hospital, Helsinki, Finland.
authors.forename: Ramon
authors.initials: R
authors.lastname: Perez Tanoira
authors.affiliation: Department of Internal Medicine, Hospital Príncipe de Asturias, Madrid, Spain.
authors.forename: Gerardo
authors.initials: G
authors.lastname: Rojo-Marcos
authors.affiliation: Department of Clinical Microbiology and Parasitology, Hospital Príncipe de Asturias, 28805, Alcalá de Henares, Madrid, Spain.
authors.forename: Peña
authors.initials: P
authors.lastname: Gómez-Herruz
authors.affiliation: Malaria and Emergent Protozoa Laboratory, National Center for Microbiology of Majadahonda, Madrid, Spain.
authors.forename: Jose Miguel
authors.initials: JM
authors.lastname: Rubio
mesh_terms: D005060:Europe; D006801:Humans; D008288:Malaria / Q000175:diagnosis* / Q000469:parasitology*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D021141:Nucleic Acid Amplification Techniques / Q000379:methods*; D041122:Plasmodium ovale / Q000235:genetics / Q000302:isolation & purification*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity
keywords: LAMP; Loop-mediated isothermal amplification; Malaria; Plasmodium ovale