Integrated Next-Generation Sequencing and qPCR Workflow in Clinical Diagnostics
Poster May 27, 2016
Elian Rakhmanaliev, Tatiana Ivanova, Gerd Michel
Sanger sequencing and PCR methods have been the standard molecular methods in clinical diagnostics for decades. NGS technology revolutionized the field of genomics, transcriptomics and metagenomics and is now swiftly becoming a routine method in different areas of clinical diagnostics. Vela Diagnostics developed an integrated automated multi-purpose Sentosa workflow, which consists of: 1) a robotic liquid handling system for nucleic acid extraction, PCR set-up and/or NGS library preparation (Sentosa SX101); 2) instruments for real-time PCR or template preparation and deep sequencing; 3) kits for nucleic acid extraction, real-time PCR-based tests, NGS library preparation assays and reagents for deep sequencing; 4) assay specific applications, and 5) data analysis and reporting software. Different diagnostic applications employ the same robotic platform for qPCR set-up and preparation of NGS libraries. Vela Diagnostics offers 7 NGS and more than 20 qPCR viral, microbial and oncology CE-IVD tests, which can be run on the same system. In addition, several extraction kits were developed to isolate nucleic acids from various types of clinical samples, including FFPE, whole blood, plasma/serum, swabs, sputum, stool and urine. These unique abilities of the Sentosa workflow provide complete and relevant information to aid clinical decision-making and patient management.
When there is a need to quickly analyze samples using a number of different PCR assays, it is likely that optimal conditions for each assay will not be the same. First, different assays often will require different annealing temperatures for their primers. In addition, amplicons may be designed to be of different lengths and therefore require varying durations of the extension step.READ MORE