HT-STELA Telomere Length Analysis via Capillary Electrophoresis
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for determining telomere length are low-resolution and unsuitable for high-throughput analysis, particularly for detecting short telomeres, which are critical for diagnosing and prognosing diseases like cancer, as they fail to reliably detect dysfunctional telomeres that can lead to genomic instability and tumorigenesis.
Innovation Solution
The development of High Throughput Single Telomere Length Analysis (HT-STELA) method using capillary electrophoresis for resolving and detecting telomere length distributions, which involves annealing primers to regions adjacent to telomere repeat arrays, PCR amplification, and optimization of DNA and cycle parameters to produce a telomere length smear for accurate analysis.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If hybridisation of DNA probes to telomere repeat units is used, then telomere length can be detected, but detection resolution deteriorates for short telomeres
Solution Approach 1:
The patent extracts the telomere repeat units from their native chromosomal context and amplifies them in vitro using PCR. This allows the telomere sequences to be isolated and analyzed separately, enabling high-resolution detection of short telomeres that would be undetectable in native chromosomal DNA using hybridisation methods
Solution Approach 2:
The patent changes the detection parameter from direct hybridisation to PCR amplification followed by fragment length analysis. By amplifying the telomere repeat units and analyzing the length of amplification products, the method achieves sufficient resolution to detect short telomeres with high reliability
2Productivity
If Q-PCR-based methods are used for high throughput analysis, then analysis throughput increases, but measurement precision deteriorates for short telomeres
Solution Approach 1:
The patent segments the analysis into two distinct stages: (1) PCR amplification of telomere repeat units from chromosomal DNA, and (2) fragment length analysis of the amplification products. This segmentation allows each stage to be optimized independently - PCR for throughput and fragment analysis for precision
Solution Approach 2:
The patent replaces the quantitative PCR measurement system with a fragment length analysis system. Instead of measuring telomere length through quantitative PCR signal intensity, the method uses direct fragment length measurement of amplified telomere products, which provides superior precision for short telomere detection while maintaining high throughput capability
3Loss of time
If current low-resolution technologies are used, then analysis time is reduced, but diagnostic value deteriorates for cancer prognosis
Solution Approach 1:
The patent performs preliminary PCR amplification of telomere repeat units before final length analysis. This preliminary action enriches the telomere sequences and creates sufficient material for high-resolution fragment length analysis, enabling both rapid processing and high diagnostic value to be achieved simultaneously
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
HT-STELA enables rapid and accurate determination of telomere lengths, reducing analysis time and increasing throughput, allowing for reliable detection of short telomeres and improved prognostic assessment of cancer patients, with results that can be obtained within a single working day.
Implementation Method 1
annealing a primer sequence to a region adjacent a telomere repeat array of chromosomal DNA
Implementation Method 2
PCR amplifying 20 ng-35 ng of said chromosomal DNA using 21-23 cycles to generate an amplification product
Implementation Method 3
The development of High Throughput Single Telomere Length_analysis (HT-STELA) method using capillary electrophoresis for resolving and detecting telomere length distributions
Data Source
AI summary
The invention relates to a high throughput method for determining telomere length of mammalian chromosomal DNA; primers for use in said method; a kit comprising said primers; use of said method to diagnose or prognose or to determine the risk of developing a telomere shortening disease such as cancer, ageing, neurological disorders including Alzheimer's disease, Parkinson's disease and other dementias, brain infarction, heart disease, chronic HIV infection, chronic hepatitis, skin diseases, chronic inflammatory bowel disease including ulcerative colitis, anaemia, atherosclerosis, Barrett's oesophagus and cancers including pre-cancerous conditions, infertility, telomere syndromes including dyskeratosis congenita, aplastic anaemia, idiopathic pulmonary fibrosis, familial myelodysplastic syndrome-acute myeloid leukaemia, Hoyeraal-Hreiderasson syndrome, Revesz syndrome, Coats plus syndrome, bone marrow failure, and cryptogenic liver cirrhosis. Additionally, the invention also has application in assessing an individual's suitability to be a transplantation donor, for example a bone marrow donor.