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Whole Genome Sequencing of Candida glabrata for Detection of Markers of Antifungal Drug Resistance
Published on: December 28, 2017
Role of ERG11 and MDR1 genes in cycloheximide and multidrug resistance in Candida species
Zill-E- Huma1, Sidrah Saleem2, Muhammad Imran2
1Department of Microbiology, University of Health Sciences, Lahore, Pakistan. zhumairfan@gmail.com.
Abstract:
Candida species are amongst the commensals of the mucosal surfaces of the human body which include the oral cavity, vagina, and intestinal mucosa. Fungal infections are on the rise worldwide. The overall burden of infections due to fungi is difficult to estimate because the majority of them remain undiagnosed. The present study aims to determine the burden of antifungal resistance in low socioeconomic country, Pakistan and the frequency of ERG11 and MDR1 genes involved. A total of 636 Candida isolates were obtained from various tertiary care institutions in Lahore in the form of culture on various culture plates. Sabouraud agar culture plates were used to culture the Candida spp. Antifungal resistance was determined against Fluconazole, Itraconazole, Ketoconazole, and Nystatin via disk diffusion technique. Most resistance was observed against Fluconazole followed by Itraconazole, Ketoconazole, and Nystatin. The Candida isolates recovering from CVP tip and tissue have a high resistance profile. Candida species resistant to at least two antifungals were chosen for further ERG11 and MDR1 detection through real-time PCR. Among 255 Candida isolates, 240 contained ERG11 gene while MDR1 gene is present in 149 Candida isolates. The isolates carrying both genes were tested by the broth microdilution technique for the susceptibility against cycloheximide, all of them were able to grow in cycloheximide. The genetic determinants of antifungal resistance such as ERG11 and MDR1 are as important in the multidrug resistance against a variety of compounds and antifungal drugs.
Insights
Antifungal resistance is rising in Pakistan, with Candida species showing high resistance to common drugs like fluconazole. Key genes ERG11 and MDR1 are frequently detected in resistant strains, highlighting their role in fungal drug resistance.
Area of Science:
- Medical Mycology
- Molecular Biology
- Infectious Diseases
Background:
- Candida species are common human commensals, but invasive fungal infections are increasing globally.
- Estimating the burden of fungal infections is challenging due to underdiagnosis.
- Antifungal resistance poses a significant threat to public health.
Purpose of the Study:
- To determine the prevalence of antifungal resistance in Candida species in Pakistan.
- To investigate the frequency of ERG11 and MDR1 genes in resistant isolates.
- To understand the genetic basis of multidrug resistance in Candida.
Main Methods:
- Collected 636 Candida isolates from tertiary care hospitals in Lahore.
- Assessed antifungal resistance using disk diffusion against fluconazole, itraconazole, ketoconazole, and nystatin.
- Detected ERG11 and MDR1 genes using real-time PCR and confirmed susceptibility via broth microdilution.
Main Results:
- High antifungal resistance observed, particularly against fluconazole, itraconazole, ketoconazole, and nystatin.
- Candida isolates from CVP tips and tissue samples exhibited elevated resistance.
- ERG11 gene was present in 240/255 isolates, and MDR1 in 149/255 isolates; both genes contribute to multidrug resistance.
Conclusions:
- Antifungal resistance is a significant issue in Pakistan, driven by genetic factors like ERG11 and MDR1.
- These genes are crucial determinants of multidrug resistance in Candida species.
- Further research is needed to combat the rising threat of antifungal resistance.
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