Use of PCR-RFLP and PCR-HWP1 for identification of Candida species isolated from cystic fibrosis patients

Background: Due to the predisposing conditions in patients with cystic fibrosis (CF) caused by defective mucociliary clearance facilitating colonization and invasion with Candida species has dramatically increased. Traditional methods for identifying problems are imminent and time-consuming. Therefo...

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Main Authors: Peyman Solimani, Samira Salari, Soheila khalizadeh, Maryam Hassanzad, Sadegh Khodavaisy, Mahdi Abastabar, Hamed Fakhim hajiaghaei, Hamid Badali
Format: Article
Language:English
Published: Mazandaran University of Medical Sciences and Health Services 2014-08-01
Series:Research in Molecular Medicine
Subjects:
Online Access:http://rmm.mazums.ac.ir/browse.php?a_code=A-10-25-4&slc_lang=en&sid=1
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author Peyman Solimani
Samira Salari
Soheila khalizadeh
Maryam Hassanzad
Sadegh Khodavaisy
Mahdi Abastabar
Hamed Fakhim hajiaghaei
Hamid Badali
author_facet Peyman Solimani
Samira Salari
Soheila khalizadeh
Maryam Hassanzad
Sadegh Khodavaisy
Mahdi Abastabar
Hamed Fakhim hajiaghaei
Hamid Badali
author_sort Peyman Solimani
collection DOAJ
description Background: Due to the predisposing conditions in patients with cystic fibrosis (CF) caused by defective mucociliary clearance facilitating colonization and invasion with Candida species has dramatically increased. Traditional methods for identifying problems are imminent and time-consuming. Therefore, molecular techniques utilizing amplification of target DNA provide quick and precise methods for the diagnosis and identification of Candida species. Therefore, the aim of the current study was to identify the most medically common isolated Candida species from the air way of CF patients by PCR-RFLP and amplification of HWP1 gene. Materials and Methods: A total of 42 CF patients presenting symptoms who referred to pediatric respiratory diseases research center were screened for the presence of Candida spp. The isolates initially were phenotypically identified and confirmed by molecular approaches based on restriction fragment length polymorphism ( PCR -RFLP) for the discrimination of C. albicans of non-albicans and the amplification of HWP1 gene for the discrimination of C. albicans from C. dubliniensis and C. africana was conducted. Results: The results show that C. albicans was the most frequently isolated species (83.8%) followed by non-albicans included C. parapsilosis (7.1%), C. glabrata (3.2%), and C. tropicalis (3.2%). The restriction patterns of each Candida species were perfectly specific. Since MspI could not discriminate between the three morphological related species, C. albicans, C. dubliniensis and C. africana, we used PCR amplification of HWP1 gene, which (7.1%) species from C. albicans identified as C. dubliniensis, however C. africana strains were not found. Conclusion: The present study found that C. albicans as predominant species wereisolated from the CF patients. It could be concluded that molecular diagnostic methods are reliable and would be useful for the identification of medically important Candida species in clinical samples . Therefore, considerable attention has been paid to the prevention and treatment of microbial growth, which has resulted in the improvement of patient management.
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spelling doaj.art-c0335c77a06f4998b6f40274696a31732022-12-21T20:31:24ZengMazandaran University of Medical Sciences and Health ServicesResearch in Molecular Medicine2322-13482322-133X2014-08-01232428Use of PCR-RFLP and PCR-HWP1 for identification of Candida species isolated from cystic fibrosis patientsPeyman Solimani0Samira Salari1Soheila khalizadeh2Maryam Hassanzad3Sadegh Khodavaisy4Mahdi Abastabar5Hamed Fakhim hajiaghaei6Hamid Badali7 1Department of Medical Mycology and Parasitology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran 1Department of Medical Mycology and Parasitology, School of Medicine, Kerman University of Medical Sciences, Kerman, Iran 2Pediatric Respiratory Diseases Research Center, NRITLD, Masih Daneshvari Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran 2Pediatric Respiratory Diseases Research Center, NRITLD, Masih Daneshvari Hospital, Shahid Beheshti University of Medical Sciences, Tehran, Iran 3Department of Medical Mycology and Parasitology, Kurdistan University of Medical Sciences, Sanandaj, Iran 5Invasive Fungi Research Centre (IFRC)/Department of Medical Mycology and Parasitology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Student Research Committee, Mazandaran University o f Medical Sciences, Sari, Iran 5Invasive Fungi Research Centre (IFRC)/Department of Medical Mycology and Parasitology, School of Medicine, Mazandaran University of Medical Sciences, Sari, Iran Background: Due to the predisposing conditions in patients with cystic fibrosis (CF) caused by defective mucociliary clearance facilitating colonization and invasion with Candida species has dramatically increased. Traditional methods for identifying problems are imminent and time-consuming. Therefore, molecular techniques utilizing amplification of target DNA provide quick and precise methods for the diagnosis and identification of Candida species. Therefore, the aim of the current study was to identify the most medically common isolated Candida species from the air way of CF patients by PCR-RFLP and amplification of HWP1 gene. Materials and Methods: A total of 42 CF patients presenting symptoms who referred to pediatric respiratory diseases research center were screened for the presence of Candida spp. The isolates initially were phenotypically identified and confirmed by molecular approaches based on restriction fragment length polymorphism ( PCR -RFLP) for the discrimination of C. albicans of non-albicans and the amplification of HWP1 gene for the discrimination of C. albicans from C. dubliniensis and C. africana was conducted. Results: The results show that C. albicans was the most frequently isolated species (83.8%) followed by non-albicans included C. parapsilosis (7.1%), C. glabrata (3.2%), and C. tropicalis (3.2%). The restriction patterns of each Candida species were perfectly specific. Since MspI could not discriminate between the three morphological related species, C. albicans, C. dubliniensis and C. africana, we used PCR amplification of HWP1 gene, which (7.1%) species from C. albicans identified as C. dubliniensis, however C. africana strains were not found. Conclusion: The present study found that C. albicans as predominant species wereisolated from the CF patients. It could be concluded that molecular diagnostic methods are reliable and would be useful for the identification of medically important Candida species in clinical samples . Therefore, considerable attention has been paid to the prevention and treatment of microbial growth, which has resulted in the improvement of patient management.http://rmm.mazums.ac.ir/browse.php?a_code=A-10-25-4&slc_lang=en&sid=1Candida species PCR-RFLP HWP1 gene cystic fibrosis
spellingShingle Peyman Solimani
Samira Salari
Soheila khalizadeh
Maryam Hassanzad
Sadegh Khodavaisy
Mahdi Abastabar
Hamed Fakhim hajiaghaei
Hamid Badali
Use of PCR-RFLP and PCR-HWP1 for identification of Candida species isolated from cystic fibrosis patients
Research in Molecular Medicine
Candida species
PCR-RFLP
HWP1 gene
cystic fibrosis
title Use of PCR-RFLP and PCR-HWP1 for identification of Candida species isolated from cystic fibrosis patients
title_full Use of PCR-RFLP and PCR-HWP1 for identification of Candida species isolated from cystic fibrosis patients
title_fullStr Use of PCR-RFLP and PCR-HWP1 for identification of Candida species isolated from cystic fibrosis patients
title_full_unstemmed Use of PCR-RFLP and PCR-HWP1 for identification of Candida species isolated from cystic fibrosis patients
title_short Use of PCR-RFLP and PCR-HWP1 for identification of Candida species isolated from cystic fibrosis patients
title_sort use of pcr rflp and pcr hwp1 for identification of candida species isolated from cystic fibrosis patients
topic Candida species
PCR-RFLP
HWP1 gene
cystic fibrosis
url http://rmm.mazums.ac.ir/browse.php?a_code=A-10-25-4&slc_lang=en&sid=1
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