Sequencing circulating miRNA in maternal plasma with modified library preparation.
ABSTRACT Circulating microRNAs in maternal plasma as one type of the cell free nucleotide acid revealed its potential for non-invasive prenatal diagnosis. The next generation sequencing technology provides promising approach detecting miRNA for such purpose.
In this present study, a modified library preparation method for SOLiD sequencing technology was developed and maternal plasma miRNA from single and twin pregnancies was analyzed. Quantitative PCR was carried out for comparison.
Results showed that the sequenced data was improved remarkably with this modified library preparation method; different types and levels of miRNA expression were found in twin pregnancy compared with control. Several miRNAs were validated that remarkably changed in twin pregnancy.
It is indicated that miRNAs might involve the process of pregnancy such as the generation of twin pregnancy, and it also suggested that the specific miRNAs could act as potential biomarkers for clinical diagnosis and therapy.
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ABSTRACT: MicroRNAs (miRNAs) are increasingly envisaged as biomarkers for various tumor and non-tumor diseases. MiRNA biomarker identification is, as of now, mostly performed in a candidate approach, limiting discovery to annotated miRNAs and ignoring unknown ones with potential diagnostic value. Here, we applied high-throughput SOLiD transcriptome sequencing of miRNAs expressed in human peripheral blood of patients with lung cancer. We developed a bioinformatics pipeline to generate profiles of miRNA markers and to detect novel miRNAs with diagnostic information. Applying our approach, we detected 76 previously unknown miRNAs and 41 novel mature forms of known precursors. In addition, we identified 32 annotated and seven unknown miRNAs that were significantly altered in cancer patients. These results demonstrate that deep sequencing of small RNAs bears high potential to quantify miRNAs in peripheral blood and to identify previously unknown miRNAs serving as biomarker for lung cancer.Molecular BioSystems 12/2011; 7(12):3187-99. · 3.35 Impact Factor
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ABSTRACT: Breast cancer treatment is improving due to the introduction of new drugs, guided by molecular testing of the primary tumour for mutations/oncogenic drivers (e.g. HER2 gene amplification). However, tumour tissue is not always available for molecular analysis, intra-tumoural heterogeneity is common and the "cancer genome" is known to evolve with time, particularly following treatment as resistance develops. After resection, those patients with only residual micrometastases are likely to be cured but those with radiologically detectable overt disease are not. Thus, the discovery of blood test(s) that could (1) alert clinicians to early primary or recurrent disease and (2) monitor response to treatment could impact significantly on mortality. Towards this, we and others have focused on molecular profiling of circulating nucleic acids isolated from plasma, both cell-free DNA (cfDNA) and microRNAs, and the relationship of these to circulating tumour cells (CTCs). This review considers the utility of each as circulating biomarkers in breast cancer with particular emphasis on the bioinformatic tools available to support molecular profiling.CANCER AND METASTASIS REVIEW 10/2012; · 9.35 Impact Factor