Shrestha Dutta, Amity University Kolkata
What is Liquid biopsy?
A fluid biopsy, otherwise called liquid biopsy or fluid phase biopsy, is the examination of non-solid organic tissue, principally blood. The fluid biopsy used in the investigation is known as rapid plasma genotyping – includes a test tube filled with blood, which contains DNA that is free-floating from malignant growth cells and examining DNA for changes or other abnormalities. This method gives a “snapshot” of principal genetic abnormalities in a tumor, is a typical tool in research for testing the molecular makeup of various types of malignancy.
What is Droplet digital PCR?
Droplet Digital PCR (ddPCR) is a strategy for performing digital PCR that depends on water-oil emulsion drop innovation. ddPCR is presented as an innovation that may work with miRNA estimation, particularly in a fluid biopsy, since it has ended up being more delicate, to offer exceptionally reproducible outcomes, and to be less vulnerable to inhibitors than ordinary RT-qPCR. The ddPCR procedure depends on the separation of the sample into a huge number of micro-reactions of characterized volume in the form of droplets in oil. After the PCR, every droplet may or may not contain the nucleic acid of interest, hence permitting the perfect estimation of the number of particles in the reaction with the help of Poisson’s distribution.
Detection of MLH1 promoter methylation by liquid biopsy and droplet digital PCR:
MutL Homolog 1 (MLH1) promotor methylation is related to microsatellite instability-high colorectal cancer (CRC). The solid relationship between’s methylation status and cancer improvement and movement has prompted a developing interest in the utilization of methylation markers in circulating tumor DNA (ctDNA) for early malignant growth identification and longitudinal checking. As cancer explicit DNA methylation changes in body liquids are restricted, it is especially difficult to apply clinical fluid biopsy techniques with high affectability and particularity. The reason for this examination was to fabricate a fit-for-purpose methylation-sensitive restriction enzyme (MSRE) based ddPCR test to analyze MLH1 promoter methylation in ctDNA in CRC.
Primers and probes were fabricated to intensify CpG locales of the MLH1 promoter. Methylated and non-methylated control genomic DNA was sheared to impersonate ctDNA and exposed to MSRE HpaII assimilation.
The research exhibited that the fluid biopsy test for MLH1 promoter methylation identification utilizing pure quantitative ddPCR is a straightforward and profoundly delicate system that gives steady methylation detection in ctDNA. The MSRE ddPCR approach can likewise be applied to a different gene of interest where methylation examples could uncover clinically applicable data for future clinical biomarkers or potential medication development.
The created fluid biopsy test for identifying MLH1 promoter methylation by ddPCR is an advantageous and cost-effective methodology that could be effortlessly moved to a clinical setting. The basic MSRE ddPCR strategy and high affectability of the test give a dependable ctDNA methylation biomarker measure to reflect single or various DNA methylation changes starting in tumor cells.
Besides, this methodology can be applied to a different gene of interest, or malignancy signs, where methylation patterns could uncover vital clinically applicable data for future clinical biomarkers as well as for developing and implementing medicines.
Also read: Low UVB exposure can increase risk of Colorectal Cancer
Reference:
- Wang, D., O’Rourke, D., Sanchez-Garcia, J. F., Cai, T., Scheuenpflug, J., & Feng, Z. (2021). Development of a liquid biopsy based purely quantitative digital droplet PCR assay for detection of MLH1 promoter methylation in colorectal cancer patients. BMC cancer, 21(1), 797. https://doi.org/10.1186/s12885-021-08497-x
Author info:
Shrestha Dutta is a 4th-year Biotechnology Engineering Student with a great interest in Genetics, Recombinant DNA Technology, and Immunology. She is a creative scientific writer in Bioxone with an inclination towards gaining knowledge regarding various sections of Biotechnology and engaging herself in various wet lab skills. She also has a review paper published in the journal IJSER.
Reference links:
- https://www.ijser.org/researchpaper/Unfaltering-boon-of-Nanotechnology-on-Plant-Growth.pdf
- https://bioxone.in/news/worldnews/therapy-for-congenital-myasthenia-a-destructive-neuromuscular-disorder/
- https://bioxone.in/news/indianews/first-cadaveric-liver-transplantation-in-india-by-hope-pump/
- https://bioxone.in/news/worldnews/nanodecoys-from-special-lung-cells-can-kill-sars-cov2/
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