A new single-step quantitative pathogen detection system: template-tagging followed by multiplex asymmetric PCR using common primers and CE-SSCP.

摘要: Rapid diagnosis of bacterial infection is important for patient management and appropriate therapy during the early phase bacteria-induced disease. Among existing techniques identifying microbial, CE-SSCP combined with 16S ribosomal RNA gene-specific PCR has benefits excellent sensitivity, resolution, reproducibility. However, even though can separate products high-resolution, multiplex detection quantification are complicated by primer-dimer formation non-specific amplification. Here, we describe a novel technique pathogens template-tagging followed asymmetric subsequent CE-SSCP. More specifically, reverse transcribed RNAs from seven septicemia-inducing pathogens, tagged templates common end sequences, amplified them using primers. The resulting amplicons could be successfully separated quantified comparison to an internal standard. This method yielded results that illustrate potential this system diagnosing infectious