An Identification of Staphylococcus spp. by PCR-Restriction Fragment Length Polymorphism (PCR-RFLP) Analysis of dnaJ Gene and ApoI Enzyme

Abstract

Genetic methods based on Polymerase Chain Reaction-Restriction Fragment Length Polymorphism (PCR-RFLP) are widely used for microbial species determination, in this study, PCR-RFLP technique was used to target the intragenic between 16S and 23S rDNA (dnaJ gene) for rapid detection and identification of twenty Staphylococcus species isolates.

The RFLP analysis of the dnaJ gene and the digestive enzyme ApoI was designed for a rapid and accurate identification of the selected genome. In this assay, the conserved intragenic region between 16S and 23S fragments subsequently digestion of the amplicon with restriction enzymes. Therefore, dnaJ gene with digestive enzyme ApoI were studied to differentiate Staphylococcus genus.

The results confirmed that S. epidermidis, S. hominis, and S. lugdunensis had 3 bands and S. aureus 4 bands. However, the restriction enzyme not fail but there are no restrict site in there amplified genome segment for S. haemolyticus, S. xylosus and S. auricularis. Our study found that a restriction enzyme ApoI does not benefit in classifying the species of the genus Staphylococcus spp