Molecular beacons as diagnostic tools: technology and applications.
Molecular beacons are single-stranded, fluorophore-labeled nucleic acid probes that are capable of generating a fluorescent signal in the presence of target, but are dark in the absence of target. Molecular beacons allow multiplex detection of PCR products in real time in a homogeneous assay format. Real time detection is inherently quantitative and affords a greater dynamic range than end-point detection methods. Reactions in a homogeneous assay format are sealed before amplification takes place, providing improved contamination control. A single cycler/reader instrument, coupled with automated sample preparation, results in higher throughput and greater ease of use. A multiplex qualitative assay that detects Chlamydia trachomatis and Neisseria gonorrhoeae, along with an internal control, has been developed. High specificity is achieved through careful selection of primers, probes and assay conditions. Quantitative HIV, HCV, and HBV viral load assays, with sensitivities of 50 copies/ml, 20 IU/ml, and 50 copies/ml, respectively, are achievable. The viral load assays are designed to quantitate all subtype and genotype specimens equivalently. A molecular beacon assay has been designed to detect a single nucleotide polymorphism in the beta2 adrenergic receptor gene.
Abravaya
Klara
K
Molecular Diagnostics, Abbott Laboratories, Abbott Park, Illinois 60064-6014, USA.
Huff
Jeffrey
J
Marshall
Ron
R
Merchant
Barbara
B
Mullen
Carolyn
C
Schneider
George
G
Robinson
John
J
D002692:Chlamydia trachomatis / Q000235:genetics; D004252:DNA Mutational Analysis; D004269:DNA, Bacterial / Q000032:analysis*; D004279:DNA, Viral / Q000032:analysis*; D005456:Fluorescent Dyes; D016680:Genome, Bacterial; D016679:Genome, Viral; D015497:HIV-1 / Q000235:genetics; D016174:Hepacivirus / Q000235:genetics; D006515:Hepatitis B virus / Q000235:genetics; D006801:Humans; D025202:Molecular Diagnostic Techniques; D009344:Neisseria gonorrhoeae / Q000235:genetics; D015341:Nucleic Acid Probes / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D020641:Polymorphism, Single Nucleotide; D018343:Receptors, Adrenergic, beta-2 / Q000235:genetics*; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence; D019562:Viral Load
D004269:DNA, Bacterial; D004279:DNA, Viral; D005456:Fluorescent Dyes; D015341:Nucleic Acid Probes; D018343:Receptors, Adrenergic, beta-2