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Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Distinguishing human and possum faeces using PCR markers
M Devane1, B Robson, F Nourozi
1Institute of Environmental Science and Research Ltd., Christchurch, New Zealand. megan.devane@esr.cri.nz
Journal of Water and Health
|August 29, 2013
Summary
New polymerase chain reaction (PCR) markers were developed to detect human fecal pollution. A specific marker for possum feces was created to improve accuracy and avoid false positives in water quality testing.
Area of Science:
- Environmental microbiology
- Molecular biology
- Water quality assessment
Background:
- Existing polymerase chain reaction (PCR) markers for human fecal pollution detection show high false-positive rates with brush-tailed possum feces.
- This cross-reactivity can compromise accurate water quality monitoring and source tracking.
Purpose of the Study:
- To develop and validate a new PCR marker specific for brush-tailed possum feces.
- To improve the reliability of human fecal pollution detection by differentiating between human and possum sources.
Main Methods:
- Developed a novel PCR marker targeting Bacteroidales 16S ribosomal ribonucleic acid sequences specific to possums.
- Tested the specificity and sensitivity of the possum marker against a diverse range of animal feces (n=233, 11 species) and human sewage.
- Evaluated cross-reactivity with human and other animal fecal samples.
Main Results:
- The developed possum PCR marker demonstrated high sensitivity (83.3%) in possum feces (n=36).
- Specificity of the possum marker was high (95.7%), with limited cross-reactivity to black swan, human, and rabbit feces at very low concentrations.
- The possum marker was not detected in human sewage or feces from other tested animal species.
Conclusions:
- A novel, highly specific PCR marker for brush-tailed possum feces has been developed.
- This marker can be used in conjunction with human-specific markers to accurately identify fecal pollution sources in water.
- Further testing using multiple fecal source tracking methods is recommended when both human and possum markers are detected to confirm pollution origins.
