Evaluation of target-enrichment accompanied by next-generation sequencing for detection of four pathogens in bloodstream infections / 军事医学

ABSTRACT

Objective To establish an Ampliseq method that combines target-enrichment and the next-generation sequencing for simultaneous detection of Staphylococcus aureus, S.epidermidis, Klebsiella pneumoniae and Listeria monocytogenes in order to provide a fast and accurate means to detect pathogens in bloodstream infections.Methods A method to evaluate the LOD,specificity and sensitivity by constructing simulated samples spiked in known pathogens was established.Results and Conclusion Target-enrichment Ampliseq showed good sensitivity and specificity,and the limit of detection(LOD)was as low as 101CFU/ml.The sensitivity was 95.38%,the specificity was 95.45%and the Kappa value was between 0.839 -1.000.This method can detect S.aureus,S.epidermidis,K.pneumoniae and L.monocytogenes simultaneously in one reaction within 15 hours.