A DNA microarray based on Arrayed - Primer Extension ( APEX ) technique for identification 1 of pathogenic fungi responsible for invasive and superficial mycoses
نویسندگان
چکیده
27 28 An oligonucleotide microarray, based on the arrayed primer extension (APEX) technique, has been 29 developed to simultaneously identify pathogenic fungi frequently isolated from invasive and 30 superficial infections. Species-specific oligonuceotide probes complementary to the internal 31 transcribed spacer (ITS1 and ITS2) region were designed for 24 species belonging to 10 genera, 32 including Candida (albicans, dubliniensis, famata, glabrata, tropicalis, kefyr; krusei, 33 guilliermondii, lusitaniae, metapsilosis, orthopsilosis, parapsilosis, and pulcherrima), Cryptococcus 34 neoformans, Aspergillus (fumigatus and terreus), Trichophyton (rubrum and tonsurans), 35 Trichosporon cutaneum, Epidermophyton floccosum, Fusarium solani, Microsporum canis, 36 Penicillium marneffei, and Saccharomyces cerevisiae. The microarray was tested for its specificity 37 with a panel of reference and blinded clinical isolates. The APEX technique was proven to be 38 highly discriminative, leading to unequivocal identification of each species including the highly 39 related ones C. parapsilosis, C. orthopsilosis and C. metapsilosis. Because of the satisfactory basic 40 performance traits obtained, such as reproducibility, specificity and unambiguous interpretation of 41 the results, this new system represents a reliable method of potential use in clinical laboratories for 42 parallel one-shot detection and identification of the most common pathogenic fungi. 43 AC EP TE D on O cber 6, 2017 by gest ht://jcm .sm .rg/ D ow nladed fom
منابع مشابه
DNA microarray based on arrayed-primer extension technique for identification of pathogenic fungi responsible for invasive and superficial mycoses.
An oligonucleotide microarray based on the arrayed-primer extension (APEX) technique has been developed to simultaneously identify pathogenic fungi frequently isolated from invasive and superficial infections. Species-specific oligonucleotide probes complementary to the internal transcribed spacer 1 and 2 (ITS1 and ITS2) region were designed for 24 species belonging to 10 genera, including Cand...
متن کاملDevelopment of a single tube 640-plex genotyping method for detection of nucleic acid variations on microarrays
Detection of DNA sequence variation is critical to biomedical applications, including disease genetic identification, diagnosis and treatment, drug discovery and forensic analysis. Here, we describe an arrayed primer extension-based genotyping method (APEX-2) that allows multiplex (640-plex) DNA amplification and detection of single nucleotide polymorphisms (SNPs) and mutations on microarrays v...
متن کاملMACGT: multi-dimensional automated clustering genotyping tool for analysis of microarray-based mini-sequencing data
SUMMARY Multi-dimensional Automated Clustering Genotyping Tool (MACGT) is a Java application that clusters complex multi-dimensional vector data derived from single nucleotide polymorphism (SNP) genotyping experiments using mini-sequencing based microarray chemistries such as arrayed primer extension (APEX). Spot intensity output files from microarray experiments across multiple samples are imp...
متن کاملOligonucleotide Array for Mutation Analysis in Familial Breast Cancer
Detecting recurrent mutations in the BRCA1 and BRCA2 genes presents a challenge because these mutations are scattered throughout each of the genes. An efficient mutation detection system for these genes should have the properties of a scanning methodology in combination with the fidelity of DNA sequence analysis. We have evaluated an Arrayed Primer EXtension (APEX) methodology applied to the id...
متن کاملEvaluation of arrayed primer extension for TP53 mutation detection in breast and ovarian carcinomas.
Mutations in the tumor suppressor gene TP53 are associated with a wide range of different cancers and may have prognostic and therapeutic implications. Methods for rapid and sensitive detection of mutations in this gene are therefore required. In order to make screening more effective, a commercially available TP53 genotyping microarray from Asper Biotech has been constructed by arrayed primer ...
متن کامل