Using Multiplex PCR Technique for Detection of Some Pathogenic Gram Positive bacteria and Determination of Its Antibiotic Resistance

Abstract

450 samples were collected from different pathological cases (urine, wounds, Burns, stool, and nasal and pharyngeal swabs) from November 2014 through February 2015. 60 samples were diagnosed as Staph. aureus and 35 samples were Strep. pyogenes using phenotypic, cultural and biochemical diagnosis features and definitely diagnosed with Vaitek test. Results of antibiotic resistance against different antibiotics showed variations in their resistance to these antibiotics, and 10 isolates were selected of each gender on the basis of variation in antibiotic resistance. A number of primers were designed to detect bacteria use distinct gene and also to detects resistance to antibiotics using multiplex PCR. Agarose gel electrophoresis for polymerization reaction showed that all Staph. aureus isolates had femA which might be used as detection marker where all Strep. pyogenes had Tu(tuf), in the same reaction of multiplex PCR ,detection of seven genes related with resistance to different antibiotic groups (Amp, tetR, Tri, Gen, Imp, Qnl and mecA) were done, all Staph. aureus contained Imp,Gen,mecA, tetR,Tri, and Qnl genes while its varied in their contain of Amp gene . Also all Strep. pyogenes had Tri,Gen,Amp and mecA genes while its varied in their contain of Qnl which not detected in all isolates. From this study it's easy to use multiplex PCR to detect pathogenic bacteria along with their antibiotic resistance without need to long routine work.