title: Multiplatform comparison of multiplexed bead arrays using HPV genotyping as a test case.
abstract: While previous studies have investigated the utility of Luminex technology in comparison to other standard techniques, there have been few studies directly comparing different bead-based assays. A key barrier to establishing Luminex technology in research or clinical laboratories is the apparent need to purchase not only encoded bead sets but also the Luminex instrument. However, as flow cytometry instrumentation continues to improve in sensitivity and in the number and diversity of detection parameters, a diverse range of bead-based assays is likely to emerge. Human papillomavirus (HPV) genotyping requires multiplexed analysis of 10-100 individual genotypes per sample, which is well suited to bead-based assays whilst technically challenging and costly for related technologies (e.g., qPCR). Here we performed an unbiased technical comparison between Luminex technology and our in-house 3-mercaptopropyl trimethoxysilane ("MPS") bead platform, which has been designed for integration with generic cytometry instruments. In genotyping 200 clinical samples, we compared the two bead assays against the goldstandard Roche Line Blot (RLB) assay, and both performed well in receiver-operator characteristic (ROC) curve analysis. We also show instrument-based differences are a significant factor in comparing the methods, which needs to be considered in future comparative studies. These multi-platform analyses are important in establishing the validity of new methods, as well as highlighting specific advantages and disadvantages of the assays for specific applications.
authors.affiliation: HPV Research Laboratory, Department of Pathology, School of Medicine, University of Washington, Seattle, USA.
authors.forename: Simon R
authors.initials: SR
authors.lastname: Corrie
authors.forename: Qinghua
authors.initials: Q
authors.lastname: Feng
authors.forename: Tiffany
authors.initials: T
authors.lastname: Blair
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Hawes
authors.forename: Nancy B
authors.initials: NB
authors.lastname: Kiviat
authors.forename: Matt
authors.initials: M
authors.lastname: Trau
mesh_terms: D052685:Alphapapillomavirus / Q000235:genetics* / Q000302:isolation & purification; D004279:DNA, Viral / Q000235:genetics; D005260:Female; D005434:Flow Cytometry / Q000295:instrumentation / Q000379:methods*; D005453:Fluorescence; D005838:Genotype; D015658:HIV Infections / Q000150:complications / Q000821:virology; D006801:Humans; D008163:Luminescent Measurements; D008863:Microspheres; D009693:Nucleic Acid Hybridization; D030361:Papillomavirus Infections / Q000150:complications / Q000821:virology; D010957:Plasmids / Q000032:analysis; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D002583:Uterine Cervical Neoplasms / Q000150:complications / Q000821:virology
chemical_list: D004279:DNA, Viral