Abstract
Gliotoxin is an important virulence factors in Aspergillus fumigatus. The biosynthesis of this mycotoxin is regulated and expressed by the presence of gliP genes. This study aimed to identify Aspergillus fumigatus isolates in clinical and environmental sources with glip genes using conventional PCR and sequence. To achieve this, DNA was isolated from twenty A. fumigatus isolates using commercial kit. The range of the DNA extracted was 65-210 ng/μl with a purity of 1.5-1.9. Species identification of the A. fumigatus isolates was achieved to a high specificity by using tailored primer. The results showed that all isolates had positive results to the primer and all isolates were able to produce gliotoxin. PCR detected the gliotoxin genes, glip in five isolates. The five PCR product samples were sent for sequence analysis and 25 µl (10 pmol) from the forward primer. The results of all the samples indicated have a single band of the desired product of gliP gene of A.fumigatus and the samples sent for sequencing related to molecular weight 190 bp.
Highlights
[1] A. fumigatus cause pulmonary clinical forms of Aspergillosis disease spatially in immunocompromised patients or those undergoing immunosuppressive therapy prior to organ transplantation and may cause invasive disease, the forms of pulmonary clinical forms are sap rophytic, allergic and invasive, invasive aspergillosis (IA) is the most risky form of the disease, since it involves the invasion of viable tissue and may produce a mortality rate of 40–90% in immunosuppressed patients [2,3,4 and 5 ]
ZR Fungal/Bacterial DNA MiniPrepTM,After extraction Namedrop used to determine the purity and concentration of extracted genomic DNA and integrity was detected by running 0.8% agarose gel electrophoresis followed by staining with ethidium bromide and visualization under UV light [13]. key word The concentration and purity of the isolated DNA samples were measured by the NanoDrop spectrophotometer before the performance of Polymerase Chain Reaction (PCR), for DNA isolated by the commercial kit technique and by the manual technique
Polymerase Chain Reaction (PCR) was done for the detection of gliP gene region in 20 isolates of clinical and environmental A. fumigatus, all sample show positive for gliP gene region with PCR product 190bp, as show in figure (2)
Summary
انًؼضٔنت يٍ يظبدسAspergillus fumigatus فً فطشGliotoxin حخببغ انحًض انُٕٔي نـ جٍُبث سشٌشٌت ٔبٍئٍت فً انؼشاق. ٌُٔظى انخخهٍك انحٍٕي نٓزا انسًٕو انفطشٌت ٌٔؼبش ػُّ ٔجٕد انجٍُبث.Aspergillus fumigatus ً ْٕ ػبيم سًً يٓى فGliotoxin انخمهٍذيpcr ببسخخذاوGlip فً انسلالاث انسشٌشٌت ٔانبٍئٍت يغ جٍُبثA. fumigatus ْذفج ْزِ انذساست إنى انخؼشف ػهى ػضلاث.Glip ٍ كبٌ انؼبئذ ي. ببسخخذاو ػذة انخجبسٌتA. fumigatus حى ػضل انحًض انُٕٔي يٍ ػششٌٍ ػٍُت، نخحمٍك رنك. ٔاسسهج نخحهٍم انخسهسم إنىA. fumigatus حى انخؼشف ػهى الإَٔاع نؼضل.1.1-1.6 يٍكشٔنخش يغ َمبء/ َبَٕغشاو210-56 انحًض انُٕٔي انًسخخشجت فً َطبق ٔأظٓشث انُخبئج أٌ خًس.Afumi ٔأظٓشث انُخبئج أٌ جًٍغ انؼضلاث كبَج َخبئج إٌجببٍت ػهى ببدء. خظٕطٍت ػبنٍت ببسخخذاو ببدء خبص ٔأظٓشث. فً جًٍغ انؼضلاثGlip ٔ Gliotoxin حى انكشف ػٍ ٔجٕد جٍُبثpcr ببسخخذاوGliotoxin ػضلاث كبَج لبدسة ػهى إَخبج ً انؼٍُبث أسسهج نهخسهسم انًخؼهمت انٕصٌ انجضٌئA. نخحهٍم انخسهسمpcr حى إسسبل خًست ػٍُبث يٍ انًُخج110 bp
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
