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Paper Information

Journal:   JOURNAL OF MAZANDARAN UNIVERSITY OF MEDICAL SCIENCES   FEBRUARY 2011 , Volume 20 , Number 80; Page(s) 33 To 43.
 
Paper: 

QUANTIFICATION AND OPTIMIZATION OF ASPERGILLUS FUMIGATUS DNA DETECTION IN BLOOD SAMPLE USING REAL TIME PCR

 
 
Author(s):  NABILI MOJTABA, SHOKOHI TAHEREH*, HASHEMI SOTEH MOHAMAD BAGHER, JAN BABAIE GHASEM, AGHILI SEYED REZA, HOSEINIKHAH ZAHRA
 
* DEPARTMENT OF PARASITOLOGY & MYCOLOGY, FACULTY OF MEDICINE, CELLULAR AND MOLECULAR RESEARCH CENTER, MAZANDARAN UNIVERSITY OF MEDICAL SCIENCES, SARI, IRAN
 
Abstract: 

Background and purpose: Invasive aspergillosis (IA) is a serious opportunistic infection caused by various species of Aspergillus in immnucompromissed individuals. Basically, rapid and early diagnosis prevents IA progression. The purpose of this study was to quantify and optimize detection of Aspergillus fumigatus DNA in blood samples using Real Time PCR.
Materials and methods: Five milliliter blood samples of healthy volunteers were spiked with various dilutions of conidial suspension of Aspergillus fumigatus (106 to 101). DNA was extracted from blood using glass beads and QI Amp DNA Blood Mini Kit. The primers and hybridization probes were designed to amplify the 18S rRNA genes using the Light Cycler system and Fluorescence Resonance Energy Transfer.
Results: All PCR reactions were negative with other species of fungal DNA with 100% specificity. Melting curve analysis showed the species- specific melting temperature patterns on 69.14oC which helps differentiating Aspergillus fumigatus. The lower limit was determined 101 CFU (100 fg) or 10 conidia or cells per PCR reaction which indicates a high sensitivity.
Conclusion: Real-time PCR method employing hybridization probes is a highly specific and sensitive approach to determine the fungal load for early diagnosis and monitoring treatment.

 
Keyword(s): INVASIVE ASPERGILLOSIS, QUANTITATIVE PCR, DIAGNOSIS
 
 
References: 
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Cite:
APA: Copy

NABILI, M., & SHOKOHI, T., & HASHEMI SOTEH, M., & JAN BABAIE, G., & AGHILI, S., & HOSEINIKHAH, Z. (2011). QUANTIFICATION AND OPTIMIZATION OF ASPERGILLUS FUMIGATUS DNA DETECTION IN BLOOD SAMPLE USING REAL TIME PCR. JOURNAL OF MAZANDARAN UNIVERSITY OF MEDICAL SCIENCES, 20(80), 33-43. https://www.sid.ir/en/journal/ViewPaper.aspx?id=197599



Vancouver: Copy

NABILI MOJTABA, SHOKOHI TAHEREH, HASHEMI SOTEH MOHAMAD BAGHER, JAN BABAIE GHASEM, AGHILI SEYED REZA, HOSEINIKHAH ZAHRA. QUANTIFICATION AND OPTIMIZATION OF ASPERGILLUS FUMIGATUS DNA DETECTION IN BLOOD SAMPLE USING REAL TIME PCR. JOURNAL OF MAZANDARAN UNIVERSITY OF MEDICAL SCIENCES. 2011 [cited 2021April23];20(80):33-43. Available from: https://www.sid.ir/en/journal/ViewPaper.aspx?id=197599



IEEE: Copy

NABILI, M., SHOKOHI, T., HASHEMI SOTEH, M., JAN BABAIE, G., AGHILI, S., HOSEINIKHAH, Z., 2011. QUANTIFICATION AND OPTIMIZATION OF ASPERGILLUS FUMIGATUS DNA DETECTION IN BLOOD SAMPLE USING REAL TIME PCR. JOURNAL OF MAZANDARAN UNIVERSITY OF MEDICAL SCIENCES, [online] 20(80), pp.33-43. Available: https://www.sid.ir/en/journal/ViewPaper.aspx?id=197599.



 
 
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