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Whole Genome Sequencing of Candida glabrata for Detection of Markers of Antifungal Drug Resistance
Published on: December 28, 2017
Identification of Azole Resistance Markers in Clinical Isolates of Candida tropicalis Using cDNA-AFLP Method
Ali Kanani1, Farideh Zaini1, Parivash Kordbacheh1
1Department of Medical Mycology and Parasitology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran.
Background:
Global reports have highlighted the increasing prevalence of Candida tropicalis infections as well as organism(') s drug resistance. This study aimed at identifying azole resistance markers in clinical isolates of C. tropicalis, which will be a great resource for developing new drugs.
Methods:
Two susceptible and resistant isolates of C. tropicalis were recovered from an epidemiological investigation of candidiasis in immunocompromised patients. C. tropicalis ATCC 750 was used as reference strain. Antifungal susceptibility to fluconazole and itraconazole was determined using Clinical and Laboratory Standards Institute (CLSI) method. Complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) technology and real-time reverse-transcriptase (RT) PCR were used for identification of potential genes involved in azole resistance of C. tropicalis clinical isolates.
Results:
Five genes encoding the following enzymes were identified as superoxide dismutase (SOD) implicated in antioxidant defense, ornithine aminotransferase (OAT), acetyl ornithine aminotransferase (ACOAT), adenosylmethionine-8-amino-7-oxononanoate aminotransferase (DAPA AT), and 4-aminobutyrate aminotransferase (ABAT)-belonging to pyridoxal phosphate (PLP) dependent enzymes and acting in an important physiological role in many fungal-cell cycles. Real-time RT-PCR confirmed mRNA level of the aforementioned genes.
Conclusion:
Our findings showed that factors such as PLP-dependent enzymes and SOD might be implicated in drug resistance in C. tropicalis clinical isolate. Therefore, further studies are required to explore the accurate biological functions of the mentioned genes that would be helpful for effective drug development.
Insights
This study identified genes related to antioxidant defense and pyridoxal phosphate-dependent enzymes in drug-resistant Candida tropicalis. These findings offer insights for developing new antifungal drugs against Candida tropicalis infections.
Area of Science:
- Mycology
- Antimicrobial Resistance
- Molecular Biology
Background:
- Candida tropicalis infections are rising globally, accompanied by increasing drug resistance.
- Identifying azole resistance markers in C. tropicalis is crucial for developing novel antifungal therapies.
Purpose of the Study:
- To identify genetic markers associated with azole resistance in clinical isolates of Candida tropicalis.
- To provide a foundation for the development of new antifungal drugs targeting C. tropicalis.
Main Methods:
- Susceptible and resistant C. tropicalis isolates were analyzed.
- Antifungal susceptibility testing for fluconazole and itraconazole was performed using CLSI guidelines.
- Complementary DNA-amplified fragment length polymorphism (cDNA-AFLP) and real-time RT-PCR were employed to identify resistance-associated genes.
Main Results:
- Five genes were identified: superoxide dismutase (SOD), ornithine aminotransferase (OAT), acetyl ornithine aminotransferase (ACOAT), adenosylmethionine-8-amino-7-oxononanoate aminotransferase (DAPA AT), and 4-aminobutyrate aminotransferase (ABAT).
- These genes encode enzymes involved in antioxidant defense (SOD) and pyridoxal phosphate (PLP)-dependent pathways.
- Real-time RT-PCR confirmed the elevated mRNA levels of these identified genes.
Conclusions:
- Pyridoxal phosphate-dependent enzymes and superoxide dismutase may play a role in azole drug resistance in C. tropicalis.
- Further research into the specific biological functions of these genes is necessary for effective antifungal drug development.
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