AMPLIFICATION OF SALMONELLA CHROMOSOMAL DNA USING THE POLYMERASE CHAIN-REACTION

A Salmonella-specific polymerase chain reaction (PCR) was developed an d standardized. The origin of the primers was a recombinant clone (C7) that contained Salmonella-specific HindIII fragment DNA of 2.1-kiloba se pairs. Based on the sequence data of Salmonella enteritidis recombi nant clone C7, two primers designated NK1 (21 nucleotides) and NK2 (24 nucleotides) were synthesized for use in the PCR. A Salmonella-specif ic 2.0-kilobase pair DNA product was amplified by the primers from 23 species of Salmonella, but not from 19 enteric and non-enteric bacteri a. As little as 330 fg of Salmonella DNA was detected using either eth idium bromide/ultraviolet exposure of gels or Southern blot hybridizat ion with a C7 clone.