Serum miRNA profiling by quantitative RT PCR [RT-PCR Array 2]

Initial screening, the expression of 125 mature miRNA was compared between pooled control and autism samples by two microarray analysis. The differential expression of 14 miRNA was further validated by SYBR green quantitative PCR of Individual samples. Thirteen miRNAs were differetially expressed in autistic individuals compared to the controls (8 upregulated and 5 down regulated). miR-151a-3p, miR-181b-5p, miR-320a, miR-328, miR-433, miR-489, miR-572, and miR-663a were downregulated, while miR-101-3p, miR-106b-5p, miR-130a-3p, miR-195-5p, and miR-19b-3p were upregulated. We have identified a set of serum miRNAs that could be used as non-invasive biomarkers for autism. RNA from age- and gender- matched control (n=40) and autism (n=40) serum samples were pooled separately to generate four pools per group, with each pool consisting of 10 samples. miRNAs were detected and quantified using SYBR Green real-time quantitative PCR performed on ABI PRISM 7900 Sequence Detection System (SDS) [Applied Biosystems (ABI), Foster City, CA, USA]. This observed an altered expression of 14 miRNAs in the autism samples compared to that of healthy controls.