RESUMEN
BACKGROUND: Many bacterial infections are associated with biofilm formation. It is one of the important virulent factors of E. coli in urinary tract, causing recurrent and drug resistant infections. Fecal E. coli colonize the urethra and spread up the urinary tract to the bladder and kidney. Type 1 fimbriae are surface located adhesion organelles of E. coli that are directly associated with adherence to the urinary tract. The present study was aimed to study biofilm production in E. coli isolated from urinary tract infection and to correlate it with expression of fimH gene and compare its sequences. METHOD: Total 150 urine samples were processed for isolation and identification of uropathogens. E. coli isolates were further processed for detection of biofilm by TCP method and screened for the presence of fimH gene by PCR using specific primers. The PCR products were purified and sequenced bidirectionally by Sanger dideoxy sequencing system using ABI 3500 Genetic analyzer. RESULTS: From the total 98 urine samples with significant bacteriuria, 77 E. coli were isolated out of which, 40 were positive for in vitro biofilm production. Among them11 were classified as strong biofilm producers and 29 as moderate. The fimH gene from E. coli isolates was amplified using specific primers and appeared as a band of about 508bp on agarose gel. It was noted that the fimH gene was detected in moderate and strong biofilm forming E. coli while absent in non biofilm isolates.The sequences showed 99% similarity with fim H gene of E coli. CONCLUSION: The high binding ability of fimH could result in increased bacterial binding to target cells and increased pathogenicity of E. Coli.
RESUMEN
Current protocols for isolation of genomic DNA from Terminalia arjuna have their own limitations due to the presence of high content of gummy polysaccharides and polyphenols. DNA isolated by these protocols is contaminated with a yellowish, sticky and viscous matrix. In our modified DNA isolation method polysaccharides and polyphenols are removed prior to the precipitation of the DNA. Then the genomic DNA was precipitated using isopropanol. This protocol yielded a high molecular weight DNA isolated from fresh as well as dry leaves of T. arjuna, which was free from contamination and colour. Isolated DNA can be used for restriction digestion and PCR amplification. The main objective of the present protocol is to provide a simple method of isolation of DNA, using in house prepared reagents.