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

Genyan Patrick Yang; Dean D Erdman; Maria L Tondella; Barry S Fields

doi:10.1016/j.jviromet.2009.08.004

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

J Virol Methods. 2009 Dec;162(1-2):288-90. doi: 10.1016/j.jviromet.2009.08.004. Epub 2009 Aug 20.

Authors

Genyan Patrick Yang¹, Dean D Erdman, Maria L Tondella, Barry S Fields

Affiliation

¹ Centers for Disease Control and Prevention, Division of Bacterial Diseases, Respiratory Diseases Branch, Atlanta, GA 30333, USA. [email protected]

PMID: 19699237
DOI: 10.1016/j.jviromet.2009.08.004

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.

Publication types

Evaluation Study

MeSH terms

Bacteria / classification
Bacteria / genetics
Bacteria / isolation & purification
Bacterial Infections / virology
DNA Probes*
Fluorescent Dyes* / classification
Fluorescent Dyes* / metabolism
Humans
Reagent Kits, Diagnostic*
Respiratory System* / microbiology
Respiratory System* / virology
Reverse Transcriptase Polymerase Chain Reaction / methods*
Rhodamines* / metabolism
Taq Polymerase
Virus Diseases / virology
Viruses / classification
Viruses / genetics
Viruses / isolation & purification

Substances

DNA Probes
Fluorescent Dyes
Reagent Kits, Diagnostic
Rhodamines
tetramethylrhodamine
Taq Polymerase