Detection and identification of Ehrlichia species in blood by use of PCR and electrospray ionization mass spectrometry.
Rapid detection and identification of Ehrlichia species improves clinical outcome for patients suspected of ehrlichiosis. We describe an assay that employs multilocus PCR and electrospray ionization mass spectrometry (PCR/ESI-MS) to detect and identify Ehrlichia species directly from blood specimens. The results were compared to those of a colorimetric microtiter PCR enzyme immunoassay (PCR-EIA) used as a diagnostic assay. Among 213 whole-blood samples collected from patients who were clinically suspected of ehrlichiosis from 1 May to 1 August 2008 at Vanderbilt University Hospital, 40 were positive for an Ehrlichia species by PCR/ESI-MS, giving a positive rate of 18.8%. In comparison to the PCR-EIA, PCR/ESI-MS possessed a sensitivity, a specificity, and positive and negative predictive values of 95.0%, 98.8%, 95.0%, and 98.8%, respectively. The 38 specimens that were positive for Ehrlichia by both PCR/ESI-MS and the PCR-EIA were further characterized to the species level, with 100% agreement between the two assays. In addition, Rickettsia rickettsii was detected by PCR/ESI-MS from four specimens that were confirmed retrospectively by serology and PCR-EIA. In three specimens, the PCR/ESI-MS assay identified Pseudomonas aeruginosa, Neisseria meningitidis, and Staphylococcus aureus; these were confirmed by culture and/or clinical diagnosis as being clinically relevant. From specimen processing to result reporting, the PCR/ESI-MS assay can be completed within 6 h, providing another laboratory tool for the diagnosis of ehrlichiosis. Moreover, this system may provide rapid detection and identification of additional pathogens directly from blood specimens.
Eshoo
Mark W
MW
Ibis Biosciences, Carlsbad, California 92008, USA.
Crowder
Chris D
CD
Li
Haijing
H
Matthews
Heather E
HE
Meng
Shufang
S
Sefers
Susan E
SE
Sampath
Rangarajan
R
Stratton
Charles W
CW
Blyn
Lawrence B
LB
Ecker
David J
DJ
Tang
Yi-Wei
YW
D016470:Bacteremia / Q000175:diagnosis* / Q000382:microbiology; D001431:Bacteriological Techniques / Q000379:methods*; D001769:Blood / Q000382:microbiology*; D004536:Ehrlichia / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D016873:Ehrlichiosis / Q000175:diagnosis* / Q000382:microbiology; D006801:Humans; D009345:Neisseria meningitidis / Q000302:isolation & purification; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D011550:Pseudomonas aeruginosa / Q000302:isolation & purification; D012284:Rickettsia rickettsii / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D013211:Staphylococcus aureus / Q000302:isolation & purification; D013997:Time Factors