Techniques like quantitative polymerase chain reaction (qPCR) can generate data in a relatively shorter time. Because of this, The United States Environmental Protection Agency (USEPA) is interested in using quantitative polymerase chain reaction (qPCR) techniques to provide better prevention of waterborne diseases. Before the use of this technique, USEPA recommends feasibility studies to be carried out to ensure proper identification of various chemicals in the water that can interfere with the qPCR technique. Previous research has shown types of qPCR interference in quantifying enterococci fecal bacteria in water. For example, studies have suggested inhibiting agents such as tannic acid act by both binding to DNA and limiting available templates and inhibiting the polymerase reaction. Our interest is in investigating Coastal Georgia beach water to identify chemicals that may affect the efficiency of the qPCR performance. This study is being conducted in collaboration with the Department of Public Health at Georgia Southern University to provide water samples and qPCR data. The coastal locations include Jekyll island, Simon island and Tybee island. Water samples were collected from January to August in 2020 from a total of 16 different sites from these 3 islands. We hope that our data will provide guidelines that will eventually help better monitor the water quality of beaches, real-time data reports to beach managers for advisory, and better prevention of waterborne diseases in a timely manner in the state of Georgia. Collected beach water from various locations were acidified, diluted, and filtered before the measurement. Our current focus is detecting and quantifying Tannic acid in the beach water via HPLC (High-Performance Liquid Chromatography) technique. We are also investigating the effect of tannic acid in qPCR performances. Seasonal variation is also a contributing factor to the growth of fecal indicator bacteria in coastal beach water. We will analyze the factors controlling the growth of bacteria as well as chemical interference that affects the qPCR data.

Assessment of interfering chemicals to detect fecal indicator bacteria in coastal Georgia beach water by the qPCR technique

ACS Spring 2022

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