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Laboratory experience and guidelines for avoiding false positive polymerase chain reaction results
T Victor1, A Jordaan, R du Toit
1Department of Medical Physiology and Biochemistry, Faculty of Medicine, University of Stellenbosch, Tygerberg, Republic of South Africa.
European Journal of Clinical Chemistry and Clinical Biochemistry : Journal of the Forum of European Clinical Chemistry Societies
|August 1, 1993
Summary
False positive polymerase chain reaction (PCR) results are a major hurdle for diagnostic labs. This study identifies amplicon buildup as the main contamination source and offers protocols to prevent it, aiding routine PCR adoption.
Area of Science:
- Clinical Microbiology
- Molecular Diagnostics
- Infectious Disease Detection
Background:
- Polymerase chain reaction (PCR) is widely used for infectious disease diagnosis but not routinely implemented in diagnostic laboratories.
- A significant barrier to PCR adoption is the frequent occurrence of false-positive results, often due to contamination.
- Contamination issues hinder the reliable application of PCR in clinical settings.
Purpose of the Study:
- To investigate the primary sources of PCR contamination in a hospital laboratory setting.
- To evaluate and recommend effective protocols for preventing false-positive PCR results.
- To facilitate the routine implementation of PCR-based diagnostic methods.
Main Methods:
- The study focused on PCR diagnostics for heat-labile enterotoxin-producing E. coli, M. tuberculosis, M. paratuberculosis, and human papillomavirus.
- Identified amplicon buildup during amplification as the main source of contamination.
- Evaluated physical and chemical decontamination protocols, including photo-induced psoralen and uracil-N-glycosylase (UNG) enzymatic systems.
Main Results:
- Amplicon accumulation during the PCR process was confirmed as the principal cause of laboratory contamination.
- Physical and chemical control measures were detailed, with specific recommendations for different laboratory scenarios.
- The uracil-N-glycosylase system proved effective for laboratories initiating diagnostic PCR work.
Conclusions:
- Implementing robust contamination control protocols is crucial for the successful integration of PCR into routine diagnostics.
- Photo-induced psoralen and uracil-N-glycosylase are effective strategies for preventing PCR contamination.
- Simple, implementable control measures can significantly improve the reliability and adoption of PCR-based diagnostic tests.