earticle

영어

Rapid and quantitative identification of pathogen has been considered as important criteria for its clinical and hygienical requirements. Among various methods, capillary electrophoresis-single strand conformation polymorphism (CE-SSCP) analysis which can separate target DNA with
sequence difference has been applied for multiplex quantitative detection of pathogen. However, polymer matrix generally used for CE-based analysis has limited resolution in CE-SSCP since it is optimized for molecular weight dependent separation. In this study, multiplex quantitative
detection of 12 pathogens using CE-SSCP was performed with noble high resolution polymer matrix. The targets were amplified with single set of universal PCR primers from conserved region of 16S rRNA gene, and the length of PCR products were 238-265 bp. A series of experiments using the different amounts of gDNAs of the pathogens exhibited 0.31 –1.56 pg of limit of detection and ~ 102 of dynamic range. The results showed that 12-plex quantitative analysis was achieved with high resolution and sensitivity, and simple procedure. The results also displayed the potential of new polymer matrix for various genetic analyses.