Detection and differentiation of avian mycoplasmas by surface-enhanced Raman spectroscopy based on a silver nanorod array.
Mycoplasma gallisepticum is a bacterial pathogen of poultry that is estimated to cause annual losses exceeding $780 million. The National Poultry Improvement Plan guidelines recommend regular surveillance and intervention strategies to contain M. gallisepticum infections and ensure mycoplasma-free avian stocks, but several factors make detection of M. gallisepticum and diagnosis of M. gallisepticum infection a major challenge. Current techniques are laborious, require special expertise, and are typically plagued by false results. In this study, we describe a novel detection strategy which uses silver nanorod array-surface-enhanced Raman spectroscopy (NA-SERS) for direct detection of avian mycoplasmas. As a proof of concept for use in avian diagnostics, we used NA-SERS to detect and differentiate multiple strains of avian mycoplasma species, including Acholeplasma laidlawii, Mycoplasma gallinarum, Mycoplasma gallinaceum, Mycoplasma synoviae, and M. gallisepticum, including vaccine strains 6/85, F, and ts-11. Chemometric multivariate analysis of spectral data was used to classify these species rapidly and accurately, with >93% sensitivity and specificity. Furthermore, NA-SERS had a lower limit of detection that was 100-fold greater than that of standard PCR and comparable to that of real-time quantitative PCR. Detection of M. gallisepticum in choanal cleft swabs from experimentally infected birds yielded good sensitivity and specificity, suggesting that NA-SERS is applicable for clinical detection.
Department of Microbiology, University of Georgia, Athens, Georgia, USA.
Suzanne L
SL
Hennigan
Jeremy D
JD
Driskell
Naola
N
Ferguson-Noel
Richard A
RA
Dluhy
Yiping
Y
Zhao
Ralph A
RA
Tripp
Duncan C
DC
Krause
D000127:Acholeplasma / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification*; D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods*; D001717:Birds / Q000382:microbiology*; D046228:Microarray Analysis / Q000379:methods; D009174:Mycoplasma / Q000737:chemistry / Q000145:classification / Q000302:isolation & purification*; D043942:Nanotubes; D012680:Sensitivity and Specificity; D012834:Silver / Q000378:metabolism*; D013059:Spectrum Analysis, Raman / Q000379:methods*
D012834:Silver