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222348

Loop-mediated isothermal amplification: A rapid and simple method for detection of fluconazole resistant <i>Candida albicans</i> vaginitis

Article

Last updated: 22 Jan 2023

Subjects

-

Tags

Antimicrobial resistance

Abstract

Background:Candidiasis is one of the most important opportunistic fungal infections. Time-saving antimicrobial susceptibility testing (AST) and molecular methods were developed to detect antimicrobial resistance. Loop-mediated isothermal amplification (LAMP) is a molecular diagnostic method performed isothermally with Bst DNA polymerase and four or six primers. Objective: Vitek identification of Candida isolates and determination of their AST were performed. However, it is time consuming; therefore, rapid and sensitive LAMP-based assay was tested as a method for phenotypic detection of resistance of Candida albicans isolates to fluconazole. Methods: Identification of Candida spp. isolated from vaginal swabs of non-pregnant women with vaginitis and minimal inhibitory concentration (MIC) values were done by Vitek2 system. Sensitive C. albicans isolates were inoculated into sabouraud´s dextrose broth without fluconazole while resistant strains were inoculated into broth with fluconazole and incubated at 37˚C. At end of 2nd, 4th and 6th h, cultures were harvested by 10 min centrifugation at 2,000 rpm and DNA was extracted by boiling. For LAMP, specific primers of C. albicans alpha-INT1 gene were used. Results: Out of 250 samples, 117 (46.8 %) were positive for Candida infections. Candida albicans isolates (57 isolates) were grouped into fluconazole-sensitive (12 isolates) and fluconazole-resistant (45 isolates). Fluconazole-resistant C. albicans could grow in presence of drug and positive LAMP reaction was obtained after a time interval similar to sensitive isolates. Conclusion: LAMP reaction allowed phenotypic detection of behavior of resistant strains in presence of antifungal agent. LAMP based evaluation of AST is superior to conventional methods and molecular detection of resistance genes.

DOI

10.21608/mid.2022.121884.1247

Keywords

Loop-Mediated Isothermal Amplification, azole resistance, Candida

Authors

First Name

Shereen

Last Name

Mohammed

MiddleName

-

Affiliation

Medical Microbiology and Immunology Department, Faculty of Medicine, Assiut University, Assiut, Egypt

Email

mokhdyd@yahoo.com

City

Assiut

Orcid

0000-0003-2327-9756

First Name

Mohamed

Last Name

Ali ElFeky

MiddleName

-

Affiliation

Professor of Medical Microbiology and Immunology, Faculty of Medicine, Assuit University

Email

mohelfeky@hotmail.com

City

Assuit

Orcid

0000-0001-6255-7400

First Name

Ahmed

Last Name

Youssef

MiddleName

Alaa

Affiliation

Department of Obstetrics and Gynecology, Faculty of Medicine, Assiut University, Assiut, Egypt

Email

ahmedalaa11282@gmail.com

City

-

Orcid

-

First Name

Rania

Last Name

Bakry

MiddleName

-

Affiliation

Department of Oncological Clinical Pathology, South Egypt Cancer Institute, Assiut University, Assiut, Egypt

Email

rbakry.md@gmail.com

City

-

Orcid

-

First Name

Yousra

Last Name

Mamdoh

MiddleName

-

Affiliation

Department of Clinical Pathology, Faculty of Medicine, Assiut University, Assiut, Egypt

Email

yousramamdoh@gmail.com

City

-

Orcid

-

First Name

Aliaa

Last Name

Ghandour

MiddleName

-

Affiliation

Medical Microbiology and Immunology Department, Faculty of Medicine, Assiut University, Assiut, Egypt

Email

aliaaghandour@aun.edu.eg

City

-

Orcid

-

Volume

3

Article Issue

2

Related Issue

32908

Issue Date

2022-05-01

Receive Date

2022-01-14

Publish Date

2022-05-01

Page Start

442

Page End

448

Print ISSN

2682-4132

Online ISSN

2682-4140

Link

https://mid.journals.ekb.eg/article_222348.html

Detail API

https://mid.journals.ekb.eg/service?article_code=222348

Order

24

Type

Original Article

Type Code

1,157

Publication Type

Journal

Publication Title

Microbes and Infectious Diseases

Publication Link

https://mid.journals.ekb.eg/

MainTitle

-

Details

Type

Article

Created At

22 Jan 2023