Identification of upper respiratory tract pathogens using electrochemical detection on an oligonucleotide microarray

Bacterial and viral upper respiratory infections (URI) produce highly variable clinical symptoms that cannot be used to identify the etiologic agent. Proper treatment, however, depends on correct identification of the pathogen involved as antibiotics provide little or no benefit with viral infection...

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Bibliographic Details
Published in:	PloS one Vol. 2; no. 9; p. e924
Main Authors:	Lodes, Michael J, Suciu, Dominic, Wilmoth, Jodi L, Ross, Marty, Munro, Sandra, Dix, Kim, Bernards, Karen, Stöver, Axel G, Quintana, Miguel, Iihoshi, Naomi, Lyon, Wanda J, Danley, David L, McShea, Andrew
Format:	Journal Article
Language:	English
Published:	United States Public Library of Science 26-09-2007 Public Library of Science (PLoS)
Subjects:	Adenoviridae - genetics Adenoviridae - isolation & purification Adenoviruses Analysis Antibiotics Assaying Bacteria Bacterial Infections - diagnosis Bacterial Infections - microbiology Biotechnology/Applied Microbiology Bordetella pertussis - genetics Bordetella pertussis - isolation & purification Chlamydophila pneumoniae - genetics Chlamydophila pneumoniae - isolation & purification Coronaviridae Coronavirus 229E, Human - genetics Coronavirus 229E, Human - isolation & purification Coronavirus OC43, Human - genetics Coronavirus OC43, Human - isolation & purification Coronaviruses DNA, Bacterial - chemistry DNA, Bacterial - genetics DNA, Viral - chemistry DNA, Viral - genetics Electrochemical analysis Electrochemistry Electrochemistry - methods Etiology Feasibility studies Gene expression Genomes Genotyping Health aspects Health care Health promotion Humans Hybridization Identification Image analysis Image processing Infections Infectious Diseases/Respiratory Infections Influenza Influenza A Influenza A virus - genetics Influenza A virus - isolation & purification Influenza B virus - genetics Influenza B virus - isolation & purification Microorganisms Molecular Biology/Bioinformatics Multiplexing Mycoplasma pneumoniae - genetics Mycoplasma pneumoniae - isolation & purification Oligonucleotide Array Sequence Analysis - methods Oligonucleotides Parainfluenza Virus 1, Human - genetics Parainfluenza Virus 1, Human - isolation & purification Parainfluenza Virus 2, Human - genetics Parainfluenza Virus 2, Human - isolation & purification Parainfluenza Virus 3, Human - genetics Parainfluenza Virus 3, Human - isolation & purification Pathogenic microorganisms Pathogens Pertussis Pneumonia Polymerase Chain Reaction Preventive medicine Probes Public Health and Epidemiology/Infectious Diseases Public Health and Epidemiology/Screening Reproducibility of Results Respiratory Medicine/Respiratory Infections Respiratory syncytial virus Respiratory Syncytial Viruses - genetics Respiratory Syncytial Viruses - isolation & purification Respiratory System - microbiology Respiratory System - virology Respiratory tract Semiconductors Sensitivity and Specificity Sequence Analysis, DNA Sexually transmitted diseases STD Streptococcus infections Streptococcus pyogenes - genetics Streptococcus pyogenes - isolation & purification Viral infections Virology/Diagnosis Virus diseases Virus Diseases - diagnosis Virus Diseases - virology Viruses Whooping cough
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Summary:	Bacterial and viral upper respiratory infections (URI) produce highly variable clinical symptoms that cannot be used to identify the etiologic agent. Proper treatment, however, depends on correct identification of the pathogen involved as antibiotics provide little or no benefit with viral infections. Here we describe a rapid and sensitive genotyping assay and microarray for URI identification using standard amplification and hybridization techniques, with electrochemical detection (ECD) on a semiconductor-based oligonucleotide microarray. The assay was developed to detect four bacterial pathogens (Bordetella pertussis, Streptococcus pyogenes, Chlamydia pneumoniae and Mycoplasma pneumoniae) and 9 viral pathogens (adenovirus 4, coronavirus OC43, 229E and HK, influenza A and B, parainfluenza types 1, 2, and 3 and respiratory syncytial virus. This new platform forms the basis for a fully automated diagnostics system that is very flexible and can be customized to suit different or additional pathogens. Multiple probes on a flexible platform allow one to test probes empirically and then select highly reactive probes for further iterative evaluation. Because ECD uses an enzymatic reaction to create electrical signals that can be read directly from the array, there is no need for image analysis or for expensive and delicate optical scanning equipment. We show assay sensitivity and specificity that are excellent for a multiplexed format.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Conceived and designed the experiments: DS AM ML WL DD. Performed the experiments: AS ML JW SM KB. Analyzed the data: DS ML JW MR. Contributed reagents/materials/analysis tools: KD MQ NI WL. Wrote the paper: AM ML DD.
ISSN:	1932-6203 1932-6203
DOI:	10.1371/journal.pone.0000924