title: Evaluation of tetramethylrhodamine and black hole quencher 1 labeled probes and five commercial amplification mixes in TaqMan real-time RT-PCR assays for respiratory pathogens.
abstract: Tetramethylrhodamine (TAMRA) and black hole quencher 1 (BHQ1) quenched probes and five one-step RT-PCR kits were evaluated in TaqMan real-time RT-PCR assays for detection of respiratory pathogens. The intra-assay variability of the BHQ1 probes were 1.2-2.8-fold lower than those of the TAMRA probes. All kits amplified the specific targets, but differed in their sensitivity by up to 3 orders of magnitude. The AgPath-ID kit provided the best overall performance for all assay targets.
authors.affiliation: Centers for Disease Control and Prevention, Division of Bacterial Diseases, Respiratory Diseases Branch, Atlanta, GA 30333, USA. gyang@cdc.gov
authors.forename: Genyan Patrick
authors.initials: GP
authors.lastname: Yang
authors.forename: Dean D
authors.initials: DD
authors.lastname: Erdman
authors.forename: Maria L
authors.initials: ML
authors.lastname: Tondella
authors.forename: Barry S
authors.initials: BS
authors.lastname: Fields
mesh_terms: D001419:Bacteria / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D001424:Bacterial Infections / Q000821:virology; D015342:DNA Probes; D005456:Fluorescent Dyes / Q000145:classification / Q000378:metabolism; D006801:Humans; D011933:Reagent Kits, Diagnostic; D012137:Respiratory System / Q000382:microbiology / Q000821:virology; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012235:Rhodamines / Q000378:metabolism; D019914:Taq Polymerase; D014777:Virus Diseases / Q000821:virology; D014780:Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification
chemical_list: D015342:DNA Probes; D005456:Fluorescent Dyes; D011933:Reagent Kits, Diagnostic; D012235:Rhodamines; C005358:tetramethylrhodamine; D019914:Taq Polymerase