Feb 5, 2008

Performance evaluation of five commercial real-time PCR reagent systems using TaqMan assays for B. anthracis detection

Clinical Biochemistry
Youvraj SohniVivek Kapur

Abstract

Real-time PCR assay sensitivity is affected by the choice and concentrations of reaction mix constituents among other factors such as primers, probes, and analytical assay platforms. Commercially available reagent mixes facilitate PCR assay set-up with fewer steps and timeliness. However, determination of analytical assay framework is important for ready-to-use real-time PCR reagent systems for rapid, quantitative and accurate detection of bioterror pathogens such as Bacillus anthracis. In this study, performance characteristics of five commercially available quantitative PCR reagent mixes were evaluated using TaqMan-based real-time PCR. The reagent systems were tested for compatibility on the ABI 7000 assay platform and compared for their distinctive analytical characteristics using the B. anthracis rpoB and pag gene real-time PCR assays. Knowledge of distinctive assay performance characteristics of commercially available qPCR reagent mixes is critical for carefully designing analytical assay systems. The ABI, ABgene and Eppendorf reagent systems performed consistently overall for the two TaqMan assays for B. anthracis detection that were used in the current study. However, the use of Eppendorf reagent system requires shorter ...Continue Reading

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References

Mentioned in this Paper

Pathologic Cytolysis
Real-Time Polymerase Chain Reaction
Buffers
Lysozyme Test
Ncbi Taxonomy
Reverse Transcriptase Polymerase Chain Reaction
Lysozyme
Muramidase
Nuclease
Technology Platform

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