MCC Gene Methylation for Basophil Identification
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Solution Overview
Problem
Current methods are inadequate for reliably and efficiently identifying and quantifying basophil granulocytes in complex samples, such as blood or tissues, due to their low abundance and similarity to other immune cells, which hinders diagnostic and prognostic applications in diseases like cancer and autoimmune disorders.
Innovation Solution
Analyzing the methylation status of specific CpG positions in the MCC gene region, particularly using bisulfite convertibility to differentiate basophil granulocytes from other cells, allowing for their identification and quantification without purification or staining, leveraging epigenetic markers and PCR-based methods.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If basophil granulocytes are identified using conventional methods in complex samples, then identification can be performed, but reliability and precision are insufficient due to low abundance and similarity to other immune cells
Solution Approach 1:
The patent changes the identification parameter from conventional morphological or functional characteristics to epigenetic methylation status of the MCC gene. By analyzing methylation patterns at specific CpG positions, the method achieves high precision and reliability in distinguishing basophils from other immune cells, even in complex samples where basophils are present in low abundance.
2Device complexity
If basophil granulocytes are identified without purification or staining, then complexity and time are reduced, but specificity and sensitivity are compromised
Solution Approach 1:
The patent replaces mechanical and chemical methods (purification, staining) with a molecular biology-based epigenetic analysis method. By using PCR-based detection of methylation status, the method achieves high specificity and sensitivity without requiring cell purification or staining steps, thereby reducing overall method complexity while maintaining or improving detection precision.
3Ease of manufacture
If conventional identification methods are used, then existing protocols can be applied, but diagnostic and prognostic value is limited
Solution Approach 1:
The patent changes the detection parameter from conventional cell surface markers or morphology to epigenetic methylation patterns. This provides new diagnostic information that is not accessible through traditional methods, enabling better characterization of basophil populations and their functional states, thereby reducing information loss while maintaining ease of application through standardized PCR-based protocols.
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
This approach enables robust and reliable identification and quantification of basophil granulocytes, providing valuable diagnostic and prognostic insights into autoimmune diseases, cancer, and immune status, with high specificity and sensitivity, even in whole blood or non-trypsinized tissues.
Implementation Method 1
analyzing a modification such as for example the methylation status of at least one CpG position in the mammalian gene region for the gene 'mutated in colorectal cancer' (MCC)
Data Source
Figure 1
AI summary
The present invention relates to a method, in particular an in vitro method, for identifying basophil granulocytes, comprising analyzing a modification such as for example the methylation status of at least one CpG position in the mammalian gene region for the gene "mutated in colorectal cancer" (MCC), wherein a demethylation or lack of modification or methylation of said gene region is indicative for a basophil granulocyte, when compared to a non-basophil granulocyte, or any other cell type in the peripheral blood or in other tissues. The analyses according to the invention can identify basophil granulocytes on an epigenetic level and distinguish them from all other cells in complex samples, such as, for example, other blood or immune cells. The present invention furthermore provides an improved method for quantifying basophil granulocytes, in particular in complex samples. The method can be performed with or without a step of purifying and/or enriching cells, preferably in whole blood and/or non-trypsinized tissue. Finally, the invention provides primers, probes and amplicons based on bisulfite treated nucleic acids.