Serum microRNA expression profile distinguishes enterovirus 71 and coxsackievirus 16 infections in patients with hand-foot-and-mouth disease

Abstract

Altered circulating microRNA (miRNA) profiles have been noted in patients with microbial infections. We compared host serum miRNA levels in patients with hand-foot-and-mouth disease (HFMD) caused by enterovirus 71 (EV71) and coxsackievirus 16 (CVA16) as well as in other microbial infections and in healthy individuals. Among 664 different miRNAs analyzed using a miRNA array, 102 were up-regulated and 26 were down-regulated in sera of patients with enteroviral infections. Expression levels of ten candidate miRNAs were further evaluated by quantitative real-time PCR assays. A receiver operating characteristic (ROC) curve analysis revealed that six miRNAs (miR-148a, miR-143, miR-324-3p, miR-628-3p, miR-140-5p, and miR-362-3p) were able to discriminate patients with enterovirus infections from healthy controls with area under curve (AUC) values ranged from 0.828 to 0.934. The combined six miRNA using multiple logistic regression analysis provided not only a sensitivity of 97.1% and a specificity of 92.7% but also a unique profile that differentiated enterovirial infections from other microbial infections. Expression levels of five miRNAs (miR-148a, miR-143, miR-324-3p, miR-545, and miR-140-5p) were significantly increased in patients with CVA16 versus those with EV71 (p<0.05). Combination of miR-545, miR-324-3p, and miR-143 possessed a moderate ability to discrimination between CVA16 and EV71 with an AUC value of 0.761. These data indicate that sera from patients with different subtypes of enteroviral infection express unique miRNA profiles. Serum miRNA expression profiles may provide supplemental biomarkers for diagnosing and subtyping enteroviral HFMD infections.

Figure 1. Differential serum expression levels of ten miRNAs in healthy control and patients with enterovirus infection.

Serum levels of miR-140-5p, miR-143, miR-148a, miR-362-3p, miR-455-5p, miR-324-3p, miR-206, and miR-628-3p except for miR-545 and miR-597 were significantly higher in patients with enterovirus infection than those in the control group. **, p<0.01.

Figure 2. Receiver operating characteristic (ROC) curve for expression changed miRNAs to distinguish enterovirus infection from healthy control.

Combination of miR-148a, miR-143, miR-324-3p, miR-628-3p, miR-130-5p, and miR-362-3p showed stronger distinguish efficiency. The AUC of 0.989 was obtained by combining above six miRNAs.

Figure 3. Differential serum expression levels of ten miRNAs in EV71 group and CVA16 group.

Serum levels of miR-148a, miR-143, miR-324-3p, miR-545, miR-140-5p, were significantly higher in CVA16 group than those in the EV71 group. *, p<0.05. **, p<0.01.

Figure 4. Receiver operating characteristic (ROC) curve for expression changed miRNAs to distinguish CVA16 infection from EV71 infection.

ROC curve of miR-545 (A), miR-324-3p (B), and miR-143 (C) showed moderate distinguish efficiency. The AUC was mild increased to 0.761 through combining above three miRNAs (D).