Identification of Pathogens Using Single/Double Strand conformation Polymorphism (SSCP/DSCP) Analysis

摘要

Single/double strand conformation polymorphism (SSCP/DSCP) analysis of polymerase chain reaction (PCR) amplified DNA fragments has emerged as a simple and sensitive screening method for the detection of small genomic variations. In this study we investigated the use of a four-color fluorescent-dye-labeled PCR-SSCP/DSCP techniqeu as a screening method to enable the identification of microorganisms. PCR-SSCP/DSCP analysis was performed on 14 kDa fusion protein-encoding gene fragments, derived fro mstrains belonging to different species of the genus Orthopoxvirus, amplified with fluorescent-dye-labeled primers. An automated gel analyzer was used for SSCP/DSCP analysis of the products. Preliminary results indicate that the PCR-SSCP/DSCP approach is a powerful method for the detection and identification of strain-specific nucleotide sequence polymorhpism within the 14 kDa protein encding gene. Therefore, it is concluded that this PCR-SSCP/DSCP technique represents a sensitive, reproducible and highly discriminatory procedure for molecular typing of pathogens.