QUANTITATIVE ASSAY FOR THE SIMULTANEOUS DETECTION AND SPECIATION OF BACTERIAL INFECTIONS

摘要

An adaptation of the real-time PCR assay allows for highly sensitive detectionof any eubacterial species with simultaneous speciation. The assay relies on a'multiprobe' design in which a single set of highly conserved sequencesencoded by the 16S rRNA gene serves as the primer pair, and it is used incombination with both an internal highly conserved sequence, the universalprobe, and an internal variable region, the species-specific probe. A pre-PCRultrafiltration step can be used to effectively decontaminate or removebackground DNA. The real-time system reliably identifies 14 common bacterialspecies with a detection limit of 50 fg. Figure 5 depicts the design ofprimers and probes for the assay.