Journal Article

Bridging a diagnostic Kawasaki disease classifier from a microarray platform to a qRT-PCR assay

Tremoulet, Adriana H., Burns, Jane C., Tempel, Dennie, van Keulen, Danielle, Hoggart, Clive J., Rodriguez-Manzano, Jesus, Kaforou, Myrsini, Habgood-Coote, Dominic, Levin, Michael, Herberg, Jethro A., Wright, Victoria J., Shimizu, Chisato, Kuiper, Rowan, Huigh, Daphne

2023

Overview

Background Kawasaki 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. Methods We 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. Results The 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. Conclusion We 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. Impact A 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.

• Publisher: Nature Publishing Group US,Nature Publishing Group
• Subject: Basic Science / Basic Science Article / Child / Child, Preschool / Diagnostic tests / Fever / Gene expression / Gene Expression Profiling / Humans / Kawasaki disease / Medical laboratories / Medicine / Medicine & Public Health / Mucocutaneous Lymph Node Syndrome - diagnosis / Mucocutaneous Lymph Node Syndrome - genetics / Pediatric Surgery / Pediatrics / Reverse Transcriptase Polymerase Chain Reaction / ROC Curve