title: Specificity and Strain-Typing Capabilities of Nanorod Array-Surface Enhanced Raman Spectroscopy for Mycoplasma pneumoniae Detection.
abstract: Mycoplasma pneumoniae is a cell wall-less bacterial pathogen of the human respiratory tract that accounts for > 20% of all community-acquired pneumonia (CAP). At present the most effective means for detection and strain-typing is quantitative polymerase chain reaction (qPCR), which can exhibit excellent sensitivity and specificity but requires separate tests for detection and genotyping, lacks standardization between available tests and between labs, and has limited practicality for widespread, point-of-care use. We have developed and previously described a silver nanorod array-surface enhanced Raman Spectroscopy (NA-SERS) biosensing platform capable of detecting M. pneumoniae with statistically significant specificity and sensitivity in simulated and true clinical throat swab samples, and the ability to distinguish between reference strains of the two main genotypes of M. pneumoniae. Furthermore, we have established a qualitative lower endpoint of detection for NA-SERS of < 1 genome equivalent (cell/μl) and a quantitative multivariate detection limit of 5.3 ± 1 cells/μl. Here we demonstrate using partial least squares- discriminatory analysis (PLS-DA) of sample spectra that NA-SERS correctly identified M. pneumoniae clinical isolates from globally diverse origins and distinguished these from a panel of 12 other human commensal and pathogenic mycoplasma species with 100% cross-validated statistical accuracy. Furthermore, PLS-DA correctly classified by strain type all 30 clinical isolates with 96% cross-validated accuracy for type 1 strains, 98% cross-validated accuracy for type 2 strains, and 90% cross-validated accuracy for type 2V strains.
authors.affiliation: Department of Microbiology, University of Georgia, Athens, GA, United States of America.
authors.forename: Kelley C
authors.initials: KC
authors.lastname: Henderson
authors.affiliation: Pneumonia Response and Surveillance Laboratory, Centers for Disease Control and Prevention, Atlanta, GA, United States of America.
authors.forename: Alvaro J
authors.initials: AJ
authors.lastname: Benitez
authors.affiliation: Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, United States of America.
authors.forename: Amy E
authors.initials: AE
authors.lastname: Ratliff
authors.affiliation: Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, United States of America.
authors.forename: Donna M
authors.initials: DM
authors.lastname: Crabb
authors.affiliation: Department of Microbiology, University of Georgia, Athens, GA, United States of America.
authors.forename: Edward S
authors.initials: ES
authors.lastname: Sheppard
authors.affiliation: Pneumonia Response and Surveillance Laboratory, Centers for Disease Control and Prevention, Atlanta, GA, United States of America.
authors.forename: Jonas M
authors.initials: JM
authors.lastname: Winchell
authors.affiliation: Department of Chemistry, University of Georgia, Athens, GA, United States of America.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Dluhy
authors.affiliation: Department of Pathology, University of Alabama at Birmingham, Birmingham, AL, United States of America.
authors.forename: Ken B
authors.initials: KB
authors.lastname: Waites
authors.affiliation: Department of Pediatrics, University of Alabama at Birmingham, Birmingham, AL, United States of America.
authors.forename: T Prescott
authors.initials: TP
authors.lastname: Atkinson
authors.affiliation: Department of Microbiology, University of Georgia, Athens, GA, United States of America.
authors.forename: Duncan C
authors.initials: DC
authors.lastname: Krause
mesh_terms: D006801:Humans; D057230:Limit of Detection; D009177:Mycoplasma pneumoniae / Q000235:genetics / Q000472:pathogenicity*; D043942:Nanotubes; D011019:Pneumonia, Mycoplasma / Q000382:microbiology*; D012680:Sensitivity and Specificity; D013059:Spectrum Analysis, Raman / Q000379:methods*