Amplification of Proteus mirabilis chromosomal DNA using the polymerase chain reaction

A Proteus mirabilis-specific polymerase chain reaction (PCR) was developed and standardized. The origin of the primers was a recombinant clone that co ntained P. mirabilis-specific Hind III fragment DNA of 3.5-kilobase pairs. Based on the sequence data of P. mirabilis recombinant clone, two primers d esignated MMKAP 1 and MMKAP 2 were synthesized for use in the PCR. A P. mir abilis-specific 3.5-kb pair DNA product was amplified by the primers from 1 8 strains of P. mirabilis, but not from other Protease species and bacteria . The minimum amount of target DNA detected by P. mirabilis PCR was 10 fg u sing ethidium bromide/ultraviolet exposure of gels or Southern blot hybridi zation with a P. mirabilis recombinant DNA probe. (C) 1999 Academic Press.