Target discrimination by surface-immobilized molecular beacons designed to detect Francisella tularensis.
A molecular beacon (MB) array was designed based on unique regions of the 16S rRNA of the bacterium Francisella tularensis. Nucleic acid molecular beacons undergo a spontaneous fluorogenic conformational change when they hybridize to specific complementary targets. The array was printed on aldehyde glass or hydrogel slides and evaluated for functioning in presence of complementary oligonucleotide sequences, single-nucleotide mismatch sequences and multiple nucleotide mismatch sequences. Discriminating true target from mismatched targets was found to be dependent on type, number, and location of mismatches within the beacon (i.e. located in the stem or loop regions). Optimal conditions for molecular beacon deposition, and target hybridization were determined for oligonucleotide target mismatch discrimination. The beacon array was stable upon recharging by exposure to an alkaline solution, and repeatedly used. In addition, performance of the beacon array biosensor was compared with molecular beacons in homogeneous solution.
Ramachandran
Akhilesh
A
Department of Physiological Sciences, College of Veterinary Medicine, Oklahoma State University, Stillwater, OK 74078, USA.
Flinchbaugh
James
J
Ayoubi
Patricia
P
Olah
Glenn A
GA
Malayer
Jerry R
JR
D000327:Adsorption; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005604:Francisella tularensis / Q000235:genetics / Q000302:isolation & purification*; D005819:Genetic Markers / Q000235:genetics; D017404:In Situ Hybridization, Fluorescence / Q000295:instrumentation* / Q000379:methods; D015347:RNA Probes / Q000032:analysis / Q000235:genetics; D012336:RNA, Ribosomal, 16S / Q000032:analysis* / Q000235:genetics; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D005819:Genetic Markers; D015347:RNA Probes; D012336:RNA, Ribosomal, 16S