Mousazade Moghadam M, Babavalian H, Mirnejad R, Shakeri F,
Volume 6, Issue 1 (spring-summer[PERSIAN] 2012)
Abstract
Abstract
Background and objectives: Genomic DNA extraction of bacterial cells is of processes performed normally in most biological laboratories therefore, various methods have been offered, manually and kit, which may be time consuming and costly. In this paper, genomic DNA extraction of Staphylococcus aureus was investigated using some laundry detergent brands available in Iran to achieve a rapid and cost effective method.
Material and Methods: five-enzyme Taj brand, three-enzyme Saftlan brand ,and Darya and Pak brands without enzyme were used in the concentrations of 10, 20, 40, 80 mg/L. Afterwards, in order to evaluate the efficiency of extracted DNA in downstream processing, PCR test was performed for femA gene in the genome of Staphylococcus aureus.
Results: DNA extraction using different concentrations of the brands show that extracted DNA using 40 mg/L Saftlan and Taj brand powders have the best results according concentration (µg/ml) and purity (A260/A280) parameters. These parameters are 387.5 1.88 (Taj), 254.1 2.80 (Softlan), 396.6 1.95 (Manual) and 423.3 2.2 (Kit), respectively. Afterward, the PCR test results by show that DNA extraction using laundry detergents has no effect on its efficiency in order to be used in downstream processes.
Conclusion: These results indicate that the proper concentrations of laundry detergents can be used to extract genomic DNA with similar efficiency to kit and manual extraction methods.
Key words: Bacterial genome, DNA extraction, laundry powder, PCR, Staphylococcus aureus
Babaii Kochaksaraii M, Nasrolahiomran A, Javid N, Shakeri F, Yazdi M, Ghaemi E A,
Volume 6, Issue 1 (spring-summer[PERSIAN] 2012)
Abstract
Abstract Background and objectives: The increase of ESBL producing E.coli can create a tremendous difficult y for the health system. These isolates leads to rapid transmission of causative genes to other clinically important bacteria and synchronously increased resistant to other antibiotics. This study was carried out to determine the prevalence of this isolate and related genes in Gorgan, North of Iran. Material and Methods: This study was conducted on 218 isolated E.coli from urinary tract infection of outpatients referring to six medical laboratories in Gorgan, during 2010-11. The resistance to Cefotaxim (Mast Co.) was assessed by Kirby-Bauer disk diffusion method. The confirmatory test for detection of resistant isolates was carried out by double disk method at the presence of Cefotaxim and clavulanic acid. The presence of β lactamase gene of blatem, blactx and blashv in ESBL was assessed by PCR method. Results: of 218, 70 isolates (32.1%) are resistant to Cefotaxim. Sixty-two (88.6%) of them are confirmed as ESBL producing E.coli. β lactamase genes of blactx, blatem and blashv can be seen in 28(45.2%), 26(41.9%) and 6(9.7%) isolates, respectively. Conclusion: the prevalence of ESBL producing E.coli in Gorgan is in the range of country average and blaCTX-M gene is the most common gene. Key words: E.coli, ESBL, bla gene, UTI
