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Quality Controls for Pcr

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Abstract:The purpose of this presentation is to present an overview of the quality control (QC) sections of a draft EPA document entitled, "Quality Assurance/Quality Control Guidance for Laboratories Performing PCR Analyses on Environmental Samples." This document has been prepared by the Office of Water and the Office of Research and Development to serve as guidance for environmental sample analyses involving PCR techniques, for incorporation into new EPA PCR-based methods, for use with EPA grants and assistance agreements involving PCR methods and as guidance to evaluate PCR data in technical papers. This presentation will describe when and how to use four positive controls and two negative controls with PCR methods and will describe the proper controls for assays used to confirm PCR results. The four positive controls consist of a "PCR Positive Control," which is used to verify that reagents have been prepared properly; the "Inhibition Positive Control," which is used to show that interfering constituents of the environmental matrix have been removed adequately and do not interfere with the PCR portion of the method; the "Method Positive Control," which is used to demonstrate that the entire method (sampling, concentration, inhibitor removal, PCR, confirmation) is working; and the "Matrix Spike," which gives confidence that the matrix is not interfering with recovery and detection of target organisms over the entire method. The negative controls consist of a "PCR Negative Control," which shows that the PCR reagents do not contain contaminating substances; and a "Method Blank," which verifies that contamination has not been introduced throughout the processing and assay steps. Procedures for reducing QC failures and for corrective actions will be described.
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Citation:Fout, G. S. Quality Controls for Pcr. Presented at Water Quality Technology Conference, Philadelphia, PA, November 2-5, 2003.
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Contact: Mary P. O'Bryant - (919)-541-4871 or obriant.mary@epa.gov
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Division: Microbiological & Chemical Exposure Assessment Division
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Branch: Biohazard Assessment Research Branch
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Product Type: Abstrct/Oral
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Presented: 11/02/2003
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Related Entries:
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Bullet Item Detecting Ccl-Related, Emerging Waterborne Human Viruses and Viral Indicators for Exposure Assessment
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Last Updated on Monday, October 22, 2007
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