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181186

Multiplex PCR - RFLP Assay for Identification of Cryptococcus neoformans and Cryptococcus gattii Isolated from Birds’ Droppings and Eucalyptus Trees

Article

Last updated: 03 Jan 2025

Subjects

-

Tags

Academic and Pre-clinical Veterinary Sciences (Physiology, Histology, …rology, Parasitology, Pharmacology, Forensic Medicine and Toxicology)

Abstract

Cryptococcus neoformans/ Cryptococcus gattii (C. neoformans/ C. gattii) species complex are encapsulated basidiomycetous yeasts, causing cryptococcosis which is a life-threatening fungal disease of the pulmonary and central nervous system of humans and animals.  This study aimed to investigate the recovery rate of C. neoformans and C. gattii from bird droppings and Eucalyptus trees in Egypt as well as the performance of the phenotypic and molecular identification methods for Cryptococcus species identification. Overall, 27 Cryptococcus isolates (13.5%) were isolated from 200 examined samples including 70 pigeon droppings, 50 captive birds' droppings, and 80 Eucalyptus trees samples. The recoveredisolates were phenotypically identified based on macro- and micro-morphological characters, urease test, and differentiation using cryptococcus differential agar media. Multiplex polymerase chain reaction (PCR) followed by restriction fragment length polymorphism (RFLP) analysis using EcoRIrestriction enzyme for confirmation of species identification. The molecular methods identified 20 C. neoformansfrom pigeon's droppings (12/70, 17.14%) and captive birds' droppings samples (8/50, 16%), as well as 7 C. gattii from Eucalyptus trees (7/80, 8.75%). Molecular identification results did not correspond with those of the phenotypic identification methods in three isolates (11.11%), as phenotypic methods identified only 4 C. gattii isolates and molecular methods identified 7 isolates. In conclusion, multiplex PCR and RFLP analysis of multiplex PCR products are rapid, sensitive, species-specific, and more reliable methods for identification of Cryptococcus species and may be used as a complementary to phenotypic methods to avoid false-negative and false-positive results.

DOI

10.21608/zvjz.2021.79965.1139

Keywords

C. neoformans, C. gattii, identification, RFLP, Multiplex PCR

Authors

First Name

Mohamed

Last Name

Taha

MiddleName

-

Affiliation

Microbiology Department, Faculty of Veterinary Medicine, Zagazig University 44511, Zagazig, Egypt

Email

mtahalab@yahoo.com

City

-

Orcid

-

First Name

Yasmine

Last Name

Tartor

MiddleName

-

Affiliation

Microbiology Department, Faculty of Veterinary Medicine, Zagazig University 44511, Zagazig, Egypt

Email

jasmen21@yahoo.com

City

Zagazig

Orcid

0000-0003-1246-6548

First Name

Sara

Last Name

Ibrahim

MiddleName

-

Affiliation

Veterinarian, Faculty of Veterinary Medicine, Zagazig University 44511, Zagazig, Egypt

Email

vetsarazaki@gmail.com

City

zagazig

Orcid

-

Volume

49

Article Issue

2

Related Issue

25927

Issue Date

2021-06-01

Receive Date

2021-06-19

Publish Date

2021-06-01

Page Start

181

Page End

193

Print ISSN

1110-1458

Online ISSN

2357-075X

Link

https://zvjz.journals.ekb.eg/article_181186.html

Detail API

https://zvjz.journals.ekb.eg/service?article_code=181186

Order

5

Type

Original Article

Type Code

601

Publication Type

Journal

Publication Title

Zagazig Veterinary Journal

Publication Link

https://zvjz.journals.ekb.eg/

MainTitle

Multiplex PCR - RFLP Assay for Identification of Cryptococcus neoformans and Cryptococcus gattii Isolated from Birds’ Droppings and Eucalyptus Trees

Details

Type

Article

Created At

22 Jan 2023