Rapid Identification of Allergenic and Pathogenic Molds in Environmental Air by an Oligonucleotide Array

Overview

Journal BMC Infect Dis

Publisher Biomed Central

Specialty Infectious Diseases

Date 2011 Apr 14

PMID 21486490

Citations 4

Authors

Wen-Tsung Hung

Shu-Li Su

Lin-Yi Shiu

Tsung C Chang

Affiliations

Soon will be listed here.

Abstract

Background: Airborne fungi play an important role in causing allergy and infections in susceptible people. Identification of these fungi, based on morphological characteristics, is time-consuming, expertise-demanding, and could be inaccurate.

Methods: We developed an oligonucleotide array that could accurately identify 21 important airborne fungi (13 genera) that may cause adverse health problems. The method consisted of PCR amplification of the internal transcribed spacer (ITS) regions, hybridization of the PCR products to a panel of oligonucleotide probes immobilized on a nylon membrane, and detection of the hybridization signals with alkaline phosphatase-conjugated antibodies.

Results: A collection of 72 target and 66 nontarget reference strains were analyzed by the array. Both the sensitivity and specificity of the array were 100%, and the detection limit was 10 pg of genomic DNA per assay. Furthermore, 70 fungal isolates recovered from air samples were identified by the array and the identification results were confirmed by sequencing of the ITS and D1/D2 domain of the large-subunit RNA gene. The sensitivity and specificity of the array for identification of the air isolates was 100% (26/26) and 97.7% (43/44), respectively.

Conclusions: Identification of airborne fungi by the array was cheap and accurate. The current array may contribute to decipher the relationship between airborne fungi and adverse health effect.

Citing Articles

Comparison of the BluePoint MoldID oligonucleotide array and Bruker Biotyper MALDI-TOF MS for the identification of filamentous fungi.

Chou C, Chao Q, Lu Y, Lee T, Hsueh P, Huang Y J Clin Microbiol. 2024; 63(1):e0104824.

PMID: 39636116 PMC: 11784249. DOI: 10.1128/jcm.01048-24.

PCR-Based Microarray Enhances Diagnosis of Culture-Negative Biopsied Tissue in Patients with Invasive Mold Infections: Real-World Experience in a Tertiary Medical Center.

Jan H, Tsai C, Cia C, Lee C, Chen Y, Lee N J Fungi (Basel). 2024; 10(8).

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Comparison of DNA Microarray, Loop-Mediated Isothermal Amplification (LAMP) and Real-Time PCR with DNA Sequencing for Identification of Fusarium spp. Obtained from Patients with Hematologic Malignancies.

Souza M, Matsuzawa T, Sakai K, Muraosa Y, Lyra L, Busso-Lopes A Mycopathologia. 2017; 182(7-8):625-632.

PMID: 28324245 DOI: 10.1007/s11046-017-0129-5.

Isolation and drug susceptibility of Candida parapsilosis sensu lato and other species of C. parapsilosis complex from patients with blood stream infections and proposal of a novel LAMP identification method for the species.

Trabasso P, Matsuzawa T, Fagnani R, Muraosa Y, Tominaga K, Resende M Mycopathologia. 2014; 179(1-2):53-62.

PMID: 25481844 DOI: 10.1007/s11046-014-9830-9.

Identification of fungal pathogens by visible microarray system in combination with isothermal gene amplification.

Sakai K, Trabasso P, Moretti M, Mikami Y, Kamei K, Gonoi T Mycopathologia. 2014; 178(1-2):11-26.

PMID: 24952715 PMC: 4098066. DOI: 10.1007/s11046-014-9756-2.

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