Comparison of fluorescence and resonance light scattering for highly sensitive microarray detection of bacterial pathogens.
Microarrays have emerged as potential tools for bacterial detection and identification. Given their high parallelism, they might represent a breakthrough in current diagnostic methods, provided they can be coupled to simplified labeling protocols and detected with adequate sensitivities. We describe here a technique to directly label total bacterial RNA, thus avoiding the multiple steps and possible biases associated with enzymatic amplification (e.g. PCR). We have then compared the performances of one white-light source and two laser-based fluorescence scanners for detection reliability and sensitivity. Our study reveals that nanoparticle-labeled bacterial RNA generates reproducible resonance light scattering signals that are at least 50 times more intense than state-of-the-art confocal-based fluorescence signals.
Francois
Patrice
P
Genomic Research Laboratory, Division of Infectious Diseases, University Hospitals of Geneva, CH-1211 Geneva 14, Switzerland. patrice.francois@genomic.ch
Bento
Manuela
M
Vaudaux
Pierre
P
Schrenzel
Jacques
J
D008027:Light; D008856:Microscopy, Fluorescence; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012329:RNA, Bacterial / Q000737:chemistry / Q000235:genetics; D015203:Reproducibility of Results; D012542:Scattering, Radiation; D012680:Sensitivity and Specificity; D013211:Staphylococcus aureus / Q000235:genetics* / Q000302:isolation & purification
D012329:RNA, Bacterial