title: Comparative analysis of two broad-range PCR assays for pathogen detection in positive-blood-culture bottles: PCR-high-resolution melting analysis versus PCR-mass spectrometry.
abstract: Detection of pathogens in bloodstream infections is important for directing antimicrobial treatment, but current culture-based approaches can be problematic. Broad-range PCR assays which target conserved genomic motifs for postamplification amplicon analysis permit detection of sepsis-causing pathogens. Comparison of different broad-range assays is important for informing future implementation strategies. In this study, we compared positive-blood-culture bottles processed by PCR coupled to high-resolution melting curve analysis (PCR/HRMA) and PCR coupled to electrospray ionization-mass spectrometry (PCR/ESI-MS) to microbiology culture results. Genus-level concordance was 90% (confidence interval [CI], 80 to 96%) for PCR/HRMA and 94% (CI, 85 to 98%) for PCR/ESI-MS. Species-level concordance was 90% (CI, 80 to 96%) for PCR/HRMA and 86% (CI, 75 to 93%) for PCR/ESI-MS. Unlike PCR/HRMA, PCR/ESI-MS was able to resolve polymicrobial samples. Our results demonstrated that the two assays have similar overall concordance rates but may have different roles as potential adjunctive tests with standard blood culture, since each method has different capabilities, advantages, and disadvantages.
authors.affiliation: Johns Hopkins University, Baltimore, MD, USA. kyj2@duke.edu
authors.forename: Kevin
authors.initials: K
authors.lastname: Jeng
authors.forename: Charlotte A
authors.initials: CA
authors.lastname: Gaydos
authors.forename: Lawrence B
authors.initials: LB
authors.lastname: Blyn
authors.forename: Samuel
authors.initials: S
authors.lastname: Yang
authors.forename: Helen
authors.initials: H
authors.lastname: Won
authors.forename: Heather
authors.initials: H
authors.lastname: Matthews
authors.forename: Donna
authors.initials: D
authors.lastname: Toleno
authors.forename: Yu-Hsiang
authors.initials: YH
authors.lastname: Hsieh
authors.forename: Karen C
authors.initials: KC
authors.lastname: Carroll
authors.forename: Justin
authors.initials: J
authors.lastname: Hardick
authors.forename: Billy
authors.initials: B
authors.lastname: Masek
authors.forename: Alexander
authors.initials: A
authors.lastname: Kecojevic
authors.forename: Rangarajan
authors.initials: R
authors.lastname: Sampath
authors.forename: Stephen
authors.initials: S
authors.lastname: Peterson
authors.forename: Richard E
authors.initials: RE
authors.lastname: Rothman
mesh_terms: D001769:Blood / Q000382:microbiology*; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008828:Microbiological Techniques / Q000379:methods*; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D009691:Nucleic Acid Denaturation; D010865:Pilot Projects; D016133:Polymerase Chain Reaction / Q000379:methods*; D012189:Retrospective Studies; D012680:Sensitivity and Specificity; D018805:Sepsis / Q000175:diagnosis* / Q000382:microbiology*; D044366:Transition Temperature