title: The use of PCR/Electrospray Ionization-Time-of-Flight-Mass Spectrometry (PCR/ESI-TOF-MS) to detect bacterial and fungal colonization in healthy military service members.
abstract: The role of microbial colonization in disease is complex. Novel molecular tools to detect colonization offer theoretical improvements over traditional methods. We evaluated PCR/Electrospray Ionization-Time-of-Flight-Mass Spectrometry (PCR/ESI-TOF-MS) as a screening tool to study colonization of healthy military service members.
  
  
  
  We assessed 101 healthy Soldiers using PCR/ESI-TOF-MS on nares, oropharynx, and groin specimens for the presence of gram-positive and gram-negative bacteria (GNB), fungi, and antibiotic resistance genes. A second set of swabs was processed by traditional culture, followed by identification using the BD Phoenix automated system; comparison between PCR/ESI-TOF-MS and culture was carried out only for GNB.
  
  
  
  Using PCR/ESI-TOF-MS, at least one colonizing organism was found on each individual: mean (SD) number of organisms per subject of 11.8(2.8). The mean number of organisms in the nares, groin and oropharynx was 3.8(1.3), 3.8(1.4) and 4.2(2), respectively. The most commonly detected organisms were aerobic gram-positive bacteria: primarily coagulase-negative Staphylococcus (101 subjects: 341 organisms), Streptococcus pneumoniae (54 subjects: 57 organisms), Staphylococcus aureus (58 subjects: 80 organisms) and Nocardia asteroides (45 subjects: 50 organisms). The mecA gene was found in 96 subjects. The most commonly found GNB was Haemophilus influenzae (20 subjects: 21 organisms) and the most common anaerobe was Propionibacterium acnes (59 subjects). Saccharomyces species (30 subjects) were the most common fungi detected. Only one GNB (nares E. coli) was identified in the same subject by both diagnostic systems.
  
  
  
  PCR/ESI-TOF-MS detected common colonizing organisms and identified more typically-virulent bacteria in asymptomatic, healthy adults. PCR/ESI-TOF-MS appears to be a useful method for detecting bacterial and fungal organisms, but further clinical correlation and validation studies are needed.
authors.affiliation: San Antonio Military Medical Center, JBSA Fort Sam Houston, San Antonio, TX, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Vetor
authors.affiliation: San Antonio Military Medical Center, JBSA Fort Sam Houston, San Antonio, TX, USA.|Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.
authors.forename: Clinton K
authors.initials: CK
authors.lastname: Murray
authors.affiliation: San Antonio Military Medical Center, JBSA Fort Sam Houston, San Antonio, TX, USA.|Infectious Disease Clinical Research Program, Uniformed Services University of the Health Sciences, Bethesda, MD, USA.
authors.forename: Katrin
authors.initials: K
authors.lastname: Mende
authors.affiliation: Center for Genomic Sciences, Allegheny Singer Research Institute, Pittsburgh, PA, USA.
authors.forename: Rachel
authors.initials: R
authors.lastname: Melton-Kreft
authors.affiliation: Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA.|US Army Institute of Surgical Research, Fort Sam Houston, San Antonio, TX, USA.
authors.forename: Kevin S
authors.initials: KS
authors.lastname: Akers
authors.affiliation: US Army Institute of Surgical Research, Fort Sam Houston, San Antonio, TX, USA.
authors.forename: Joseph
authors.initials: J
authors.lastname: Wenke
authors.affiliation: Center for Genomic Sciences, Allegheny Singer Research Institute, Pittsburgh, PA, USA.
authors.forename: Tracy
authors.initials: T
authors.lastname: Spirk
authors.affiliation: US Army Institute of Surgical Research, Fort Sam Houston, San Antonio, TX, USA.
authors.forename: Charles
authors.initials: C
authors.lastname: Guymon
authors.affiliation: San Antonio Military Medical Center, JBSA Fort Sam Houston, San Antonio, TX, USA.|Infectious Disease Clinical Research Program, Uniformed Services University of the Health Sciences, Bethesda, MD, USA.
authors.forename: Wendy
authors.initials: W
authors.lastname: Zera
authors.affiliation: San Antonio Military Medical Center, JBSA Fort Sam Houston, San Antonio, TX, USA.
authors.forename: Miriam L
authors.initials: ML
authors.lastname: Beckius
authors.affiliation: Landstuhl Regional Medical Center, Landstuhl, Germany.
authors.forename: Elizabeth R
authors.initials: ER
authors.lastname: Schnaubelt
authors.affiliation: Center for Genomic Sciences, Allegheny Singer Research Institute, Pittsburgh, PA, USA.
authors.forename: Garth
authors.initials: G
authors.lastname: Ehrlich
authors.affiliation: San Antonio Military Medical Center, JBSA Fort Sam Houston, San Antonio, TX, USA. todd.j.vento.mil@mail.mil.|Uniformed Services University of the Health Sciences, Bethesda, Maryland, USA. todd.j.vento.mil@mail.mil.|Infectious Disease Service, (MCHE-MDI), Brooke Army Medical Center, 3551 Roger Brooke Drive, JBSA Fort Sam Houston, 78234, Texas, USA. todd.j.vento.mil@mail.mil.
authors.forename: Todd J
authors.initials: TJ
authors.lastname: Vento
mesh_terms: D000328:Adult; D001419:Bacteria / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D004926:Escherichia coli / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D005260:Female; D005658:Fungi / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D006090:Gram-Negative Bacteria / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D006262:Health; D006801:Humans; D008297:Male; D008828:Microbiological Techniques / Q000379:methods; D064307:Microbiota; D008889:Military Personnel; D025202:Molecular Diagnostic Techniques / Q000379:methods; D010865:Pilot Projects; D016133:Polymerase Chain Reaction / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013211:Staphylococcus aureus / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification; D055815:Young Adult
keywords: Bacterial; Colonization; Electrospray ionization time-of-flight mass spectrometry; Fungal; Military; Molecular diagnostics; PCR