1. Bridging a diagnostic Kawasaki disease classifier from a microarray platform to a qRT-PCR assay
- Author
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Kuiper, Rowan, Wright, Victoria J, Habgood-Coote, Dominic, Shimizu, Chisato, Huigh, Daphne, Tremoulet, Adriana H, van Keulen, Danielle, Hoggart, Clive J, Rodriguez-Manzano, Jesus, Herberg, Jethro A, Kaforou, Myrsini, Tempel, Dennie, Burns, Jane C, and Levin, Michael
- Subjects
Biomedical and Clinical Sciences ,Clinical Sciences ,Biotechnology ,Detection ,screening and diagnosis ,4.1 Discovery and preclinical testing of markers and technologies ,4.2 Evaluation of markers and technologies ,Infection ,Child ,Humans ,Child ,Preschool ,Mucocutaneous Lymph Node Syndrome ,Reverse Transcriptase Polymerase Chain Reaction ,Gene Expression Profiling ,Fever ,ROC Curve ,Paediatrics and Reproductive Medicine ,Public Health and Health Services ,Pediatrics ,Paediatrics - Abstract
BackgroundKawasaki disease (KD) is a systemic vasculitis that mainly affects children under 5 years of age. Up to 30% of patients develop coronary artery abnormalities, which are reduced with early treatment. Timely diagnosis of KD is challenging but may become more straightforward with the recent discovery of a whole-blood host response classifier that discriminates KD patients from patients with other febrile conditions. Here, we bridged this microarray-based classifier to a clinically applicable quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay: the Kawasaki Disease Gene Expression Profiling (KiDs-GEP) classifier.MethodsWe designed and optimized a qRT-PCR assay and applied it to a subset of samples previously used for the classifier discovery to reweight the original classifier.ResultsThe performance of the KiDs-GEP classifier was comparable to the original classifier with a cross-validated area under the ROC curve of 0.964 [95% CI: 0.924-1.00] vs 0.992 [95% CI: 0.978-1.00], respectively. Both classifiers demonstrated similar trends over various disease conditions, with the clearest distinction between individuals diagnosed with KD vs viral infections.ConclusionWe successfully bridged the microarray-based classifier into the KiDs-GEP classifier, a more rapid and more cost-efficient qRT-PCR assay, bringing a diagnostic test for KD closer to the hospital clinical laboratory.ImpactA diagnostic test is needed for Kawasaki disease and is currently not available. We describe the development of a One-Step multiplex qRT-PCR assay and the subsequent modification (i.e., bridging) of the microarray-based host response classifier previously described by Wright et al. The bridged KiDs-GEP classifier performs well in discriminating Kawasaki disease patients from febrile controls. This host response clinical test for Kawasaki disease can be adapted to the hospital clinical laboratory.
- Published
- 2023