Improved techniques for molecular tagging to increase accuracy of massively parallel sequencing


Researchers at TGen have developed a method for molecular tagging (barcoding) that combines the incorporation of a unique or degenerate molecular tag during the early phases of amplification with downstream informatics analysis. This method is capable of identifying variants in complex human or non-human samples of nucleic acids at extremely low fractions such as cell-free DNA in plasma or urine or other complex mixtures for example forensic samples.


Molecular tagging involves attaching a unique or degenerate oligonucleotide label to each template molecule in the early steps of sequencing library preparation, so that any low abundance signals of variant may be distinguished from noise introduced during the process. The overall goal is to improve sensitivity and accuracy of next generation sequencing for identifying, detecting and quantifying nucleic acid molecules of any given type (or carrying a variant). The concept of molecular tagging has been described as a viable approach for detection and quantification of low abundance variants in complex mixtures of nucleic acids. However, most current methods have limited efficiency in capture molecules and require high amount of input molecules to achieve adequate sensitivity and accuracy.


The molecular tagging method developed at TGen can be used for the screening and early detection of cancer, monitoring treatment response and progression, molecular stratification and assessment of clonal evolution and treatment resistance. This method will also enable the detection of any somatic mutation in circulation in non-malignant diseases. This method can be scaled for multiplexing such that sequencing of multiple genomic regions is possible using this approach, allowing for simultaneous identification, detection and quantification of multiple mutations. It is also readily customizable and can be implemented on an ad-hoc basis or developed to focus on specific scenarios (for example cancer diagnostics using a panel of genes).

Patent Information:
For Information, Contact:
Katie Bray
Intellectual Property Counsel
The Translational Genomics Research Institute
Muhammed Murtaza
Tania Contente-Cuomo
Maria De Las Nieves Perdigones Borderias