Comparison of Three Nucleic Acid Extraction Methods in the Detection of Intestinal Pathogenic Bacteria Carried by Vectors
In order to compare and select a simple,economic and efficient method for extracting nucleic acids of intestinal pathogenic bacteria carried by vectors,nucleic acids were extracted from flies and cockroaches simulated to be infected with salmonella,Shigella and Escherichia coli using three representative methods including column extraction,magnetic bead adsorption and salt precipitation,and the mass concentration of nucleic acids,Ct value measured by quantitative fluorescence PCR(qPCR)and bacterial recovery rate were systematacially compared to fully assess the extraction efficiency of the above methods.The results showed that nucleic acids were successfully extracted from the simulated samples by both column extraction and salt precipitation,and partially by magnetic bead adsorption;and the extraction efficiency of the column extraction was the highest through comparison of mass concentration of nucleic acids,the Ct value and bacterial recovery rate.In conclusion,the affordable,accessible and accurate method,such as column extraction,could be used to detect samples with low viral load,such as vector samples,in order to improve the accuracy of detection results.
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