Circulating Salivary miRNA hsa-miR-221 as Clinically Validated Diagnostic Marker for Hand, Foot, and Mouth Disease in Pediatric Patients

Abstract

Enhancements in the diagnostic capabilities using host biomarkers are currently much needed where sensitivity and specificity issues plague the diagnosis of Hand, Foot and Mouth Disease (HFMD) in pediatrics clinical samples. We investigated miRNome profiles of HFMD saliva samples against healthy children and developed miRNA-based diagnosis models. Our 6-miRNA scoring model predicted HFMD with an overall accuracy of 85.11% in the training set and 92.86% in the blinded test set of Singapore cohort. Blinded evaluation of the model in Taiwan HFMD cases resulted in 77.08% accuracy with the 6-miRNA model and 68.75% with the 4-miRNA model. The strongest predictor of HFMD in all of the panels, hsa-miR-221 was found to be consistently and significantly downregulated in all of our HFMD cohorts. This is the first study to prove that HFMD infection could be diagnosed by circulating miRNAs in patient's saliva. Moreover, this study also serves as a stepping stone towards the future development of other infectious disease diagnosis workflows using novel biomarkers.

An epidemiological surveillance of hand foot and mouth disease in paediatric patients and in community: A Singapore retrospective cohort study, 2013–2018

“…Albeit serotype dominance of HFMD largely depending on temporal and geographical factors, underlying epidemiologic indices inflicting such paradigms are still not clearly understood [ 5 ]. Although human Enterovirus A71 (HEV-A71) and Coxsackievirus A16 (CV-A16) were major causative agents of HFMD in Singapore from 2000 to 2010, severely limited epidemiological surveillance data is available from 2010 forward despite several large outbreaks [ 2 , 6 , 7 ].…”

Bioinformatics Analysis of Key micro-RNAs and mRNAs under the Hand, Foot, and Mouth Disease Virus Infection

“…At the miRNA level, several potentially useful markers have been proposed in clinical diagnosis. A survey in Singapore reported a 6-miRNA scoring model which predicts HFMD with an overall accuracy of 85.11% in the training set and 92.86% in the blinded test set, and circulating salivary miRNA hsa-miR-221 (downregulated in that work) was regarded as a highly validated marker ( Mi et al 2018 ). Song et al have applied rhesus monkey peripheral blood mononuclear cells to search DE-miRNAs, and they identified 13 novel DE-miRNAs with 2501 targets ( Song et al 2018 ).…”

“…Previous studies have explored the disordered signals in different aspects, such as inflammatory profiles in cytokine expression ( Teo et al 2018 ; Linghua et al 2019), long non-coding RNA (lncRNA) profiles ( Meng et al 2017 ), and immune cell changes ( Wang et al 2014 ). It is conceivable and supported that patients with HFMD may exhibit changes in expression profiles of microRNAs and mRNAs, mostly derived from blood samples ( Cui et al 2011 ; Hu et al 2016 ; Yee et al 2016 ; Zhu et al 2016 ; Song et al 2017 ; Hu et al 2018 ; Jia et al 2018; Li et al 2018 ; Mi et al 2018 ; Song et al 2018 ). In addition, a comprehensive understanding of HFMD-related expression profiles and identification key markers may provide substantial diagnostic and prognostic values.…”

MicroRNA Biomarkers for Infectious Diseases: From Basic Research to Biosensing

“…Indeed, miRNAs have been found in almost all biofluids, including saliva (Min et al, 2018), urine (Ben-Dov et al, 2014, breast milk (Kelleher et al, 2019), and tears (Kim et al, 2019), however, the use of these less invasive sample types would depend on the underlying disease. For example, salivary miRNAs may be useful for the diagnosis of mouth infections (Min et al, 2018) but may not be useful for infections in other areas of the body. Blood, serum, and plasma are often used for biomarker identification as peripheral samples often contain markers of localized disease.…”