Volume 14, Issue 7 (Brucellosis Supplement 2012)
J Arak Uni Med Sci 2012, 14(7): 40-48 |
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Peeridogaheh H, valinezhad Z, Pourfarzi F. Evaluation of three DNA extraction methods for detection of brucella DNA in human serum samples. J Arak Uni Med Sci 2012; 14 (7) :40-48
URL: http://jams.arakmu.ac.ir/article-1-1384-en.html
URL: http://jams.arakmu.ac.ir/article-1-1384-en.html
1- Department of Microbiology and parasitology, School of Medicine, Ardabil University of Medical Sciences
2- School of Medicine, Ardabil University of Medical Sciences
3- Department of Community Medicine, School of Medicine, Ardabil University of Medical Sciences , f.pourfarzi@arums.ac.hr
2- School of Medicine, Ardabil University of Medical Sciences
3- Department of Community Medicine, School of Medicine, Ardabil University of Medical Sciences , f.pourfarzi@arums.ac.hr
Abstract: (13581 Views)
Background: Human brucellosis is a significant public health concern in many countries, including Iran. Therefore, the development of new diagnostic techniques, with high sensitivity and minimum risk of laboratory infection are of great importance. PCR is one of the procedures which has these advantages. However, PCR efficiency is largely dependent on DNA extraction methods. In this study, we studied the efficiency of three different extraction methods of brucella DNA in serum samples.
Materials and Methods: In this experimental study, microbial suspensions were initially prepared in saline that its turbidity was equivalent to 0.5 McFarland. Human serum samples were spiked with certain concentrations of Brucella melitensis in vitro. DNA was extracted by three methods and tested by a genus-specific PCR method.
Results: Our results showed that the cinneagen kit protocol detected brucella DNA in lower serum concentrations compared with the other protocols. Cinnagen kit could detect brucella DNA in ten-fold dilution in comparison with the other two methods.
Conclusion: According to the findings of this study, cinnagen kit was the preferred assay method that yields a better sensitivity for isolation of brucella DNA in serum samples.
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