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.
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.
Yang
Genyan Patrick
GP
Centers for Disease Control and Prevention, Division of Bacterial Diseases, Respiratory Diseases Branch, Atlanta, GA 30333, USA. gyang@cdc.gov
Erdman
Dean D
DD
Tondella
Maria L
ML
Fields
Barry S
BS
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
D015342:DNA Probes; D005456:Fluorescent Dyes; D011933:Reagent Kits, Diagnostic; D012235:Rhodamines; C005358:tetramethylrhodamine; D019914:Taq Polymerase