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Fluconazole Resistance among Oral Candida Isolates from People Living with HIV/AIDS in a Nigerian Tertiary Hospital.

ABSTRACT: Oropharyngeal candidiasis, a common fungal infection in people living with HIV/AIDS (PLWHA), arises from Candida species colonizing the oral cavity. Fluconazole is the preferred treatment and is often used empirically. Few studies have investigated the prevalence of fluconazole resistance in Nigeria. This study aimed at determining the burden of fluconazole resistance among Candida species in the oral cavities of PLWHA. We sampled the oral cavities of 350 HIV-infected adults and an equal number of HIV-negative controls. Candida isolates were identified using germ tube tests, CHROMagar Candida (CHROMagar, Paris, France), and API Candida yeast identification system (BioMérieux, Marcy-l'Étoile, France). Fluconazole susceptibility was determined using the Clinical and Laboratory Standards Institute disc diffusion method. Data were analysed using SPSS version 21 (IBM, New York, NY, USA). The significance level was set at p ? 0.05. The isolation rates for Candida amongst HIV-infected subjects and controls were 20.6% and 3.4%, respectively (p < 0.001). In PLWHA, Candida albicans was most frequently isolated (81.3%) and fluconazole resistance was present in 18 (24%) of the 75 Candida isolates. Resistance to fluconazole was present in half of the non-albicans Candida isolates. Fluconazole resistance is prevalent among oral Candida isolates in PLWHA in the study area with a significantly higher rate among non-albicans Candida spp.

SUBMITTER: Osaigbovo II

PROVIDER: S-EPMC5753171 | biostudies-literature | 2017 Dec

REPOSITORIES: biostudies-literature

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Fluconazole Resistance among Oral Candida Isolates from People Living with HIV/AIDS in a Nigerian Tertiary Hospital.

Osaigbovo Iriagbonse I II Lofor Patrick V PV Oladele Rita O RO

Journal of fungi (Basel, Switzerland) 20171208 4

Oropharyngeal candidiasis, a common fungal infection in people living with HIV/AIDS (PLWHA), arises from <i>Candida</i> species colonizing the oral cavity. Fluconazole is the preferred treatment and is often used empirically. Few studies have investigated the prevalence of fluconazole resistance in Nigeria. This study aimed at determining the burden of fluconazole resistance among <i>Candida</i> species in the oral cavities of PLWHA. We sampled the oral cavities of 350 HIV-infected adults and an ...[more]

PMID: 29371583

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Project description:Background and purposeCandidiasis is referred to a group of superficial and deep-tissue fungal infections often caused by Candida albicans. The superficial infections affect the oral, oropharynx, esophagus, and vaginal mucosa. The treatment of choice for these infections is the use of azoles, such as fluconazole. However, the increased use of these antifungal agents has led to the emergence of azole-resistant isolates of C. albicans. Different mechanisms have been suggested for the development of drug resistance, such as mutations in the encoding gene ERG11. Mutations in ERG11 result in changes in the ERG11p spatial construction and reduce the affinity between the protein and azole. This study aimed to determine the susceptibility profile of C. albicans clinical isolates to fluconazole using microdilution method. The present research was also targeted toward the detection of mutations that might be related to fluconazole resistance by the amplification and sequencing of ERG11 gene.Materials and methodsThis study was conducted on a total of 216 clinical isolates obtained from Mashhad, Isfahan, and Tehran cities in Iran, during 2016-2018. The clinical isolates were identified using molecular techniques. Furthermore, minimum inhibitory concentration (MICs) was determined according to the clinical and laboratory standards institute M27-A3 and M27-S4 documents. The concentration range for fluconazole was obtained as 0.063-64 μg/ml. In the resistant strains, ERG11 genes were amplified by specific primers. Subsequently, cycle sequencing reactions were performed on purified polymerase chain reaction (PCR) products in forward and reverse directions. Finally, the results were analyzed by MEGA (version 7) and Gene Runner software (version 6.5.30).ResultsOut of 216 strains, 100 (46.3%) species were identified as C. albicans. The MIC values for fluconazole had a range of 0.125-16 μg/ml with the MIC50 and MIC90 values of 0.5 and 1 μg/ml, respectively. Totally, 41 nucleotide changes were detected among 4 resistant isolates. In this regard, 4 out of 41 mutations in codons caused changes in ERG11p; however, these mutations did not lead to fluconazole resistance.ConclusionFluconazole resistance among clinical isolates is not merely due to the changes in ERG11p. This resistance may be also related to some other mechanisms, such as the prevention of the intracellular accumulation of the antifungal agent and alteration of the target enzyme to diminish drug binding.

Project description:BackgroundThe incidence of invasive candidiasis is increasing worldwide. However, the epidemiology, antifungal susceptibility, and virulence of Candida spp. in most hospitals remain unclear. This study aimed to evaluate invasive candidiasis in a tertiary care hospital in Nanchang City, China.MethodsMALDI-TOF MS and 18S rDNA ITS sequencing were used to identify Candida strains. Randomly amplified polymorphic DNA analysis was used for molecular typing; biofilm production, caseinase, and hemolysin activities were used to evaluate virulence. The Sensititre™ YeastOne YO10 panel was used to examine antifungal susceptibility. Mutations in ERG11 and the hotspot regions of FKS1 of drug-resistant strains were sequenced to evaluate the possible mechanisms of antifungal resistance.ResultsWe obtained 110 Candida strains, which included 40 Candida albicans (36.36%), 37 C. parapsilosis (33.64%), 21 C. tropicalis (19.09%), 9 C. glabrata (8.18%), 2 C. rugose (1.82%), and 1 C. haemulonii (0.91%) isolates. At a limiting point of 0.80, C. albicans isolates could be grouped into five clusters, C. parapsilosis and C. tropicalis isolates into seven clusters, and C. glabrata isolates into only one cluster comprising six strains by RAPD typing. Antifungal susceptibility testing revealed that the isolates showed the greatest overall resistance against fluconazole (6.36%), followed by voriconazole (4.55%). All C. albicans and C. parapsilosis isolates exhibited 100% susceptibility to echinocandins (i.e., anidulafungin, caspofungin, and micafungin), whereas one C. glabrata strain was resistant to echinocandins. The most common amino acid substitutions noted in our study was 132aa (Y132H, Y132F) in the azole-resistant strains. No missense mutation was identified in the hotpot regions of FKS1. Comparison of the selected virulence factors detectable in a laboratory environment, such as biofilm, caseinase, and hemolysin production, revealed that most Candida isolates were caseinase and hemolysin producers with a strong activity (Pz < 0.69). Furthermore, C. parapsilosis had greater total biofilm biomass (average Abs620 = 0.712) than C. albicans (average Abs620 = 0.214, p < 0.01) or C. tropicalis (average Abs620 = 0.450, p < 0.05), although all C. glabrata strains were either low- or no-biofilm producers. The virulence level of the isolates from different specimen sources or clusters showed no obvious correlation. Interesting, 75% of the C. albicans from cluster F demonstrated azole resistance, whereas two azole-resistant C. tropicalis strains belonged to the cluster Y.ConclusionThis study provides vital information regarding the epidemiology, pathogenicity, and antifungal susceptibility of Candida spp. in patients admitted to Nanchang City Hospital.

Dataset Information

Fluconazole Resistance among Oral Candida Isolates from People Living with HIV/AIDS in a Nigerian Tertiary Hospital.

Publications

Fluconazole Resistance among Oral Candida Isolates from People Living with HIV/AIDS in a Nigerian Tertiary Hospital.

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