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:: Volume 19, Number 2 (2-2017) ::
JBUMS 2017, 19(2): 26-33 Back to browse issues page
Evaluation of Real-time PCR-based DNA melting method for detection of Enterococcus faecalis and Enterococcus faecium in clinical isolates
B Zeyni, MR Arabestani, R YuosefiMashof *, H Tahmasebi
Associate professor Hamadan University of Medical Sciences , yousefimash@yahoo.com
Abstract:   (2056 Views)

BACKGROUND AND OBJECTIVE: Human error and timeless in conventional techniques to identify species Enterococcus faecium and Enterococcus faecalis are two species of the pathogenic species in humans, more accurate techniques is essential. The aim of this study is to design a method of quickly and accurately using DNA melting by Real Time PCR technique to identify and separate the two species in clinical isolates.

METHODS: In this experimental study, the bacterial isolates in the Department of Microbiology Bank of Hamadan University of Medical Sciences was used. Design of primers was done by proprietary software and selecting DivIVA gene for Enterococcus faecalis and alanine racemase for Enterococcus faecium was performed. Isolates identification was evaluated by using Real Time PCR test and melting curve temperature of DNA.

FINDINGS: Susceptibility of primers designed in divIVA gene (specific for E. faecalis) and alanine-racemase gene (specific for E. faecium) was 15CFU/ml per reaction. Specificity of designed primers by using DNA melting curve analysis was 76.6 for E. faecalis and 80.93 for E. faecium which showed considerable different in comparison with another microorganism.

CONCLUSION: Using the results obtained in this study, primers designed were sensitivity and specificity for diagnosis and differentiation of Enterococcus faecium and Enterococcus faecalis species in clinical isolates.

Keywords: Enterococcus faecalis, Enterococcus faecium, Real Time PCR
Full-Text [PDF 431 kb]   (1298 Downloads)    
Type of Study: Experimental | Subject: Mi
Received: 2016/07/17 | Accepted: 2017/03/1 | Published: 2017/03/18
* Corresponding Author Address: Department of Microbiology,Hamadan University of Medical Sciences, Hamadan, Iran
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DOI: 10.22088/jbums.19.2.26


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Zeyni B, Arabestani M, YuosefiMashof R, Tahmasebi H. Evaluation of Real-time PCR-based DNA melting method for detection of Enterococcus faecalis and Enterococcus faecium in clinical isolates . JBUMS. 2017; 19 (2) :26-33
URL: http://jbums.org/article-1-6226-en.html
Volume 19, Number 2 (2-2017) Back to browse issues page
مجله علمی دانشگاه علوم پزشکی بابل Journal of Babol University of Medical Sciences
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