Journal of Pure and Applied MicrobiologyVol. 8 No. May 2014 Special Edition

Study of Three Pair Primers PCR to Detect SEC Gene in Synovial Fluid of Rheumatoid Arthritis Patients and Comparison with Elisa

Mohammad Hosein Ataee1, Gholam Hosein Alishiri2, Davoud Esmaeili3, Akram Eidei1 and Ramezan Ali Ataee4*

1Department of Biology, Tehran Science & Research Branch, Islamic Azad University, Tehran, I.R. Iran. 2Department of Rheumatology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, I.R. Iran. 3Applied Microbial Research Center, and Department of Medical Microbiology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, I.R. Iran. 4Department of Medical Microbiology, Faculty of Medicine, Baqiyatallah University of Medical Sciences, Tehran, I.R. Iran.

Received on 23 January 2014 and accepted on 04 April 2014

 

ABSTRACT

More than 20 types of S. aureus enterotoxins have been identified so far. Detection each of them is very important in clinical samples. Different detection primers pair have been designed and used. The aim of this study was to assay the efficacy of three pair primers to detect staphylococcal enterotoxin type C in synovial fluid of RF patients and compare the results with Elisa. In this study, seventy synovial fluids of RF patients were assayed. Three pair primers were used. The primers were amplified 102, 206 and 1223bp fragment respectively. The PCR products were sequenced and compared multiple alignments with reference gene. In addition, the Elisa plate was designed for detection of the SEC. The obtained data was subjected to descriptive analysis. The results showed that the three pair primers were amplified by different frequencies. The results of amplified of amplicons 1223, 206 and 102bp were 9(18%), 34(68%) and 17(44%) positive for ent C gene respectively. The results of Elisa revealed that 22 cases (44%) were positive for staphylococcal enterotoxin C. The amplicon 206bp were the most abundant probability of the amplification product. The inter gene amplicons amplification pair primers were shown more specify. The results indicated that the staphylococcal enterotoxin C gene existed in synovial fluids of RF patients. Therefore candidate specific primers to amplify 206bp fragments of ent C and Elisa method are reliable. This finding may help to detect the etiology of rheumatoid arthritis.

Keywords : Staphylococcal enterotoxin C, PCR, Elisa, Rheumatoid arthritis.