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Real-time PCR detection of male-specific coliphages
S F Y Yong1, Y F Ngeow, Y K Tong
1School of Arts and Sciences, Monash University Malaysia, Jalan Lagoon Selatan, Bandar Sunway, Petaling Jaya 46150, Selangor, Malaysia. stacey.yong@artsci.monash.edu.my
The Malaysian Journal of Pathology
|April 2, 2008
Summary
A new real-time PCR assay effectively detects male-specific DNA coliphages, crucial indicators of enteric virus contamination in water. This method offers rapid and reproducible viral contamination assessment.
Area of Science:
- Environmental microbiology
- Molecular biology
- Virology
Background:
- Male-specific coliphages serve as vital indicators for enteric virus contamination in water.
- Traditional detection methods include plaque assays and polymerase chain reaction (PCR).
Purpose of the Study:
- To develop and validate a real-time PCR assay for detecting male-specific DNA coliphages.
Main Methods:
- Utilized the M13 coliphage as a target for assay development.
- Employed real-time PCR for quantitative DNA coliphage detection.
- Performed amplification and melting curve analyses for specificity and reproducibility assessment.
Main Results:
- The developed real-time PCR assay demonstrated a reaction efficiency of 1.45.
- Achieved a detection limit of 10(-3) plaque-forming units per reaction mix.
- Confirmed high specificity and reproducibility through repeated analyses.
Conclusions:
- Real-time PCR provides a rapid and quantitative method for assessing viral contamination levels in water.
- The assay is suitable for routine environmental monitoring of water quality.
- This advancement facilitates quicker public health risk assessments related to waterborne viruses.
