Quantification and optimization of detection Aspergillus fumigatus DNA in blood sample using Real Time PCR

Nabili, M and Shokohi, T and HashemiSoteh, M.B and JanBabaie, GH and �ghili, S.R and Hoseinikhah, Z (2011) Quantification and optimization of detection Aspergillus fumigatus DNA in blood sample using Real Time PCR. Journal of Mazandaran University of Medical Sciences, 20 (80). pp. 34-43.

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Official URL: http://jmums.mazums.ac.ir/article-1-645-en.html

Abstract

Background and purpose: �nvasive aspergillosis (��) is a serious opportunistic infection caused by various species of �spergillus in immnucompromissed individuals. Basically, rapid and early diagnosis prevents �� progression. The purpose of this study was to quantify and optimize detection of �spergillus fumigatus DN� in blood samples using Real Time P�R. Materials and methods: Five milliliter blood samples of healthy volunteers were spiked with various dilutions of conidial suspension of �spergillus fumigatus (106 to 101). DN� was extracted from blood using glass beads and Q��mp DN� Blood Mini Kit. The primers and hybridization probes were designed to amplify the 18S rRN� genes using the Light�ycler system and Fluorescence Resonance �nergy Transfer. Results: �ll P�R reactions were negative with other species of fungal DN� with 100 specificity. Melting curve analysis showed the species- specific melting temperature patterns on 69.14o� which help to differentiate �spergillus fumigatus. The lower limit was determined 101 �F� (100 fg) or 10 conidia or cells per P�R reaction which indicates a high sensitivity. �onclusion: Real-time P�R method employing hybridization probes is a highly specific and sensitive approach to determine the fungal load for early diagnosis and monitoring treatment.

Item Type: Article
Uncontrolled Keywords: �nvasive aspergillosis, quantitative P�R, diagnosis
Depositing User: Unnamed user with email eprints@mazums.ac.ir
Date Deposited: 04 Jan 2018 17:04
Last Modified: 04 Jan 2018 17:04
URI: http://eprints.mazums.ac.ir/id/eprint/2612

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