Background:The dilemma of pathogens identification in patients with unidentified clinical symptoms such as fever of unknown origin exists,which not only poses a challenge to both the diagnostic and therapeutic process by itself,but also to expert physicians.Methods:In this report,we have attempted to increase the awareness of unidentified pathogens by developing a method to investigate hitherto unidentified infectious pathogens based on unbiased high-throughput sequencing.Results:Our observations show that this method supplements current diagnostic technology that predominantly relies on information derived five cases from the intensive care unit.This methodological approach detects viruses and corrects the incidence of false positive detection rates of pathogens in a much shorter period.Through our method is followed by polymerase chain reaction validation,we could identify infection with Epstein-Barr virus,and in another case,we could identify infection with Streptococcus viridians based on the culture,which was false positive.Conclusions:This technology is a promising approach to revolutionize rapid diagnosis of infectious pathogens and to guide therapy that might result in the improvement of personalized medicine.