Detection of low numbers of bacterial cells in soils and sediments by polymerase chain reaction

Article Abstract:

Polymerase chain reaction (PCR) technology applied to the study of soils and sediments enabled the detection oflow numbers of bacterial cells. However, the sensitivity of PCR detection on DNA extracted from environmental soils and sediments was low due to the presence of humic substances or other components which inhibited the polymeraseactivities or binding of primers. Dilution of these limiting substances loweredtheir concentration but it also reduced PCR detection limit.

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Rapid method for separation of bacterial DNA from humic substances in sediments for polymerase chain reaction

Article Abstract:

A rapid gel filtration method to separate DNA from extracts containing humic substances followed by polymerase chain reaction (PCR) was used. The purification of crude DNA extracts with Sephadex G-200 columns greatly reduced the inhibitory effects of humic acid and increased the sensitivity of detection by the PCR. This technique could thus be applied to sediments containing a low content of humic substances.

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PCR-restriction fragment length polymorphism method for detection of Cyclospora cayetanensis in environmental waters without microscopic confirmation

Article Abstract:

The authors have investigated Cyclospora cayetanensis in environmental water samples. They describe the development of the alternative netsted-PCR-restriction fragment length polymorphism protocol for detection of the bacterium which differentiates C. cayetanensis amplified target sequence from those that may cross-react.

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