title: Fast DNA and protein microarray tests for the diagnosis of hepatitis C virus infection on a single platform.
abstract: Hepatitis C virus (HCV) is a major cause of chronic liver disease and liver cancer, and remains a large health care burden to the world. In this study we developed a DNA microarray test to detect HCV RNA and a protein microarray to detect human anti-HCV antibodies on a single platform. A main focus of this study was to evaluate possibilities to reduce the assay time, as a short time-to-result (TTR) is a prerequisite for a point-of-care test. Significantly reducing hybridisation and washing times did not impair the assay performance. This was confirmed first using artificial targets and subsequently using clinical samples from an HCV seroconversion panel derived from a HCV-infected patient. We were able to reduce the time required for the detection of human anti-HCV antibodies to only 14 min, achieving nanomolar sensitivity. The protein microarray exhibited an analytical sensitivity comparable to that of commercial systems. Similar results were obtained with the DNA microarray using a universal probe which covered all different HCV genotypes. It was possible to reduce the assay time after PCR from 150 min to 16 min without any loss of sensitivity. Taken together, these results constitute a significant step forward in the design of rapid, microarray-based diagnostics for human infectious disease, and show that the protein microarray is currently the most favourable candidate to fill this role.
authors: Division of Pathway Medicine, College of Medicine and Veterinary Medicine, The University of Edinburgh, Chancellor's Building, Little France Crescent, Edinburgh EH16 4SB, UK.
authors: Stuart W J
authors: SW
authors: Ember
authors: Holger
authors: H
authors: Schulze
authors: Alan J
authors: AJ
authors: Ross
authors: Julie
authors: J
authors: Luby
authors: Mizanur
authors: M
authors: Khondoker
authors: Gerard
authors: G
authors: Giraud
authors: Jonathan G
authors: JG
authors: Terry
authors: Ilenia
authors: I
authors: Ciani
authors: Chaker
authors: C
authors: Tlili
authors: Jason
authors: J
authors: Crain
authors: Anthony J
authors: AJ
authors: Walton
authors: Andrew R
authors: AR
authors: Mount
authors: Peter
authors: P
authors: Ghazal
authors: Till T
authors: TT
authors: Bachmann
authors: Colin J
authors: CJ
authors: Campbell
mesh_terms: D004279:DNA, Viral / Q000302:isolation & purification; D016174:Hepacivirus / Q000302:isolation & purification*; D006526:Hepatitis C / Q000175:diagnosis*; D018937:Hepatitis C Antibodies / Q000032:analysis; D006801:Humans; D020411:Oligonucleotide Array Sequence Analysis / Q000191:economics* / Q000379:methods; D040081:Protein Array Analysis / Q000191:economics* / Q000379:methods; D012367:RNA, Viral / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D013997:Time Factors
chemical_list: D004279:DNA, Viral; D018937:Hepatitis C Antibodies; D012367:RNA, Viral