WRRC 2024 Spring Seminar

April 5, 2:00pm - 3:00pm
Mānoa Campus, Zoom Meeting

Humic Acid Fluorescence Quenching Impacts Molecular Detection of Pathogens in Water

by Dr. Tao Yan

Nucleic acids-based molecular methods have revolutionized pathogen detection in water and other environmental matrices. However, the presence of interfering substances, such as humic acid (HA), remains a constant challenge to their accuracy and reliability. In this study, we investigated the inhibitory effect of HA on the quantification in real-time polymerase chain reactions (qPCR) and determined the underlying inhibition mechanism. Our results demonstrated that the higher concentrations of HA led to an increase in threshold cycle (CT) values. However, modeling indicated that the observation could not be explained by HA inhibition of DNA polymerase or template complexation. Further laboratory analysis and modeling identified HA quenching of fluorescence signal being the primary cause of the observed decrease in fluorescence intensity and CT value reduction.

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Mānoa Campus

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Zhiyue Wang, 956-7298,

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