Rapid identification of fungal pathogens: Molecular approaches for a new millennium

TY - JOUR

T1 - Rapid identification of fungal pathogens

T2 - Molecular approaches for a new millennium

AU - Perlin, D. S.

AU - Park, S.

PY - 2001

Y1 - 2001

N2 - The increasing prevalence of invasive fungal disease in immunocompromised patients is confounded by inadequate methods for pathogen detection. Molecular methods have been developed to address this problem because conventional methods for pathogen identification often lack sensitivity, specificity and speed, and some infectious organisms are difficult to culture. Nucleic acid-based molecular approaches for identification overcome many of the deficiencies associated with conventional methods by taking advantage of genus- and species-specific DNA sequence variations. Numerous methods have been employed that exploit both large- and small-scale genomic differences. Yet, polymerase chain reaction (PCR)-based amplification and analysis of DNA sequence polymorphisms in the internal transcribed spacer 2 region of conserved ribosomal (r)RNA genes is currently the most reliable approach for fungal identification. When combined with fluorescence-based oligonucleotide detection systems, such as TaqMan and Molecular Beacons, this approach provides real-time, quantitative, high fidelity analysis capable of single nucleotide allelic discrimination. These probe systems offer rapid turn around time (< 2 h) and are suitable for high throughput, automated, multiplex-type operations that are critical for clinical diagnostic laboratories.

AB - The increasing prevalence of invasive fungal disease in immunocompromised patients is confounded by inadequate methods for pathogen detection. Molecular methods have been developed to address this problem because conventional methods for pathogen identification often lack sensitivity, specificity and speed, and some infectious organisms are difficult to culture. Nucleic acid-based molecular approaches for identification overcome many of the deficiencies associated with conventional methods by taking advantage of genus- and species-specific DNA sequence variations. Numerous methods have been employed that exploit both large- and small-scale genomic differences. Yet, polymerase chain reaction (PCR)-based amplification and analysis of DNA sequence polymorphisms in the internal transcribed spacer 2 region of conserved ribosomal (r)RNA genes is currently the most reliable approach for fungal identification. When combined with fluorescence-based oligonucleotide detection systems, such as TaqMan and Molecular Beacons, this approach provides real-time, quantitative, high fidelity analysis capable of single nucleotide allelic discrimination. These probe systems offer rapid turn around time (< 2 h) and are suitable for high throughput, automated, multiplex-type operations that are critical for clinical diagnostic laboratories.

KW - Fungal identification

KW - Genome

KW - ITS2

KW - Molecular Beacons

KW - PCR

KW - Ribosomal genes

KW - TaqMan

UR - https://www.scopus.com/pages/publications/0035195024

UR - https://www.scopus.com/pages/publications/0035195024#tab=citedBy

M3 - Review article

SN - 0954-139X

VL - 12

SP - S13-S20

JO - Reviews and Research in Medical Microbiology

JF - Reviews and Research in Medical Microbiology

IS - SUPPL. 1

ER -