title: Validation of a loop-mediated isothermal amplification assay for rapid diagnosis of pertussis infection in nasopharyngeal samples.
abstract: OBJECTIVE
  To develop and validate a novel loop-mediated amplification (LAMP) assay for rapid diagnosis (<1 hour) of whooping cough in nasopharyngeal samples versus the gold standard: real-time PCR.
  
  
  METHODS
  The study included all nasopharyngeal samples (n = 213) collected from children with clinical suspicion of pertussis admitted to Children's University Hospital Sant Joan de Déu (Barcelona, Spain) during July-December 2014. Fresh samples were routinely analyzed by real-time PCR and stored for retrospective LAMP analysis, following an easy 30 minute DNA extraction step by Chelex-100.
  
  
  RESULTS
  Performance results of the LAMP assay were: linearity, 10(5)-10(1) CFU/ml; Limit of Detection, 2 CFU/ml; precision (mean CV), 7.38%; diagnostic sensitivity, 96.55%; diagnostic specificity, 99.46%; time to detection, 12-30 minutes.
  
  
  CONCLUSIONS
  The new test was shown to be 2.5-fold faster than real-time PCR while maintaining similar levels of analytical and clinical performance. Therefore it could become a useful diagnostic tool for molecular point-of-care testing.
authors.affiliation: a Department of Molecular Microbiology , Hospital Sant Joan de Déu, University of Barcelona , Barcelona , Spain.
authors.forename: Pedro
authors.initials: P
authors.lastname: Brotons
authors.affiliation: a Department of Molecular Microbiology , Hospital Sant Joan de Déu, University of Barcelona , Barcelona , Spain.
authors.forename: Hector D
authors.initials: HD
authors.lastname: de Paz
authors.affiliation: a Department of Molecular Microbiology , Hospital Sant Joan de Déu, University of Barcelona , Barcelona , Spain.
authors.forename: Cristina
authors.initials: C
authors.lastname: Esteva
authors.affiliation: a Department of Molecular Microbiology , Hospital Sant Joan de Déu, University of Barcelona , Barcelona , Spain.
authors.forename: Irene
authors.initials: I
authors.lastname: Latorre
authors.affiliation: a Department of Molecular Microbiology , Hospital Sant Joan de Déu, University of Barcelona , Barcelona , Spain.
authors.forename: Carmen
authors.initials: C
authors.lastname: Muñoz-Almagro
mesh_terms: D000293:Adolescent; D001886:Bordetella pertussis / Q000302:isolation & purification* / Q000472:pathogenicity; D002648:Child; D002675:Child, Preschool; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D006801:Humans; D008297:Male; D021141:Nucleic Acid Amplification Techniques / Q000379:methods; D057089:Pathology, Molecular; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D014917:Whooping Cough / Q000175:diagnosis* / Q000235:genetics / Q000382:microbiology / Q000473:pathology
chemical_list: D004269:DNA, Bacterial
keywords: Bordetella pertussis; children; loop-mediated isothermal amplification; point of care; rapid diagnosis; validation