Single cell analysis of cancer cells using an improved RT-MLPA method has potential for cancer diagnosis and monitoring

• DOI: 10.1038/srep16519
• Publication Date: 2015-11-12T11:38:54Z
• Authors: L. Kvastad; B. Werne Solnestam; E. Johansson; A. O. Nygren; N. Laddach; P. Sahlén; S. Vickovic; Schirmer C. Bendi...; M. Aaserud; L. Floer; E. Borgen; C. Schwind; R. Himmelreich; D. Latta; J. Lundeberg
• Keywords: Multiplex; Single-Cell Analysis

Abstract Single cell analysis techniques have great potential in the cancer genomics field. The detection and characterization of circulating tumour cells are important for identifying metastatic disease at an early stage monitoring it. This protocol is based on transcript profiling using Reverse Transcriptase Multiplex Ligation-dependent Probe Amplification (RT-MLPA), which a specific method simultaneous multiple mRNA transcripts. Because small amount (circulating) cells, pre-amplification reaction performed after reverse transcription to generate sufficient number target molecules MLPA reaction. We designed highly sensitive detecting quantifying panel seven genes whose expression patterns associated with breast optimized single analysis. For we used fluorescence-dependent semi-quantitative involving hybridization unique barcodes array. evaluated three human lines identified gene profiles each line. Furthermore, applied confirmed heterogeneity population. Successful from blood patients supports use RT-MLPA as diagnostic tool genomics.