Particle Measuring Method for Simultaneous Nucleic Acid and Polypeptide Detection
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Solution Overview
Problem
Current methods for detecting abnormalities in cells, such as the in situ PCR method, are limited as they primarily focus on nucleic acids and fail to effectively diagnose issues caused by polypeptides or combinations of nucleic and polypeptide abnormalities, necessitating a more comprehensive approach for pathological diagnosis.
Innovation Solution
A particle measuring method that simultaneously labels and amplifies target nucleic acids and polypeptides within cells, allowing for the simultaneous measurement of both, using a flow cytometer and specific labeling substances to determine the presence of abnormalities in nucleic acids, polypeptides, or both, without the need for protease treatment, thereby reducing complexity and increasing diagnostic reliability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If only nucleic acid detection methods (in situ PCR) are used, then nucleic acid abnormalities can be detected, but polypeptide abnormalities cannot be detected
Solution Approach 1:
The patent applies multi-functionality by enabling a single detection system to simultaneously detect both nucleic acids (via PCR amplification) and polypeptides (via immunofluorescence labeling). The flow cytometer is configured to measure multiple parameters including DNA content, RNA content, and protein expression levels from the same cell population, making the system versatile for detecting various types of cellular abnormalities without requiring separate testing procedures.
2Loss of information
If multiple separate tests are performed for nucleic acid and polypeptide abnormalities, then comprehensive diagnostic information can be obtained, but diagnostic complexity and time increase
Solution Approach 1:
The patent merges multiple diagnostic functions into a single flow cytometry-based assay. The system combines PCR amplification for nucleic acid detection with immunofluorescence labeling for polypeptide detection, allowing simultaneous measurement of DNA content, RNA content, and protein expression from the same cell samples. This integrated approach eliminates the need for separate testing procedures, reducing both diagnostic time and sample requirements while maintaining comprehensive diagnostic information.
3Measurement precision
If protease treatment is applied to enable nucleic acid access, then nucleic acid amplification is improved, but polypeptide integrity is compromised
Solution Approach 1:
The patent applies preliminary action by performing nucleic acid extraction and PCR amplification on a portion of the cell sample before polypeptide detection. Alternatively, it uses gentle cell lysis methods that release nucleic acids without requiring protease treatment, preserving polypeptide integrity for subsequent immunofluorescence labeling. This preliminary separation of detection targets allows optimal conditions for each type of detection without mutual interference.
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 method enhances the reliability of pathological diagnosis by providing comprehensive information on nucleic acid and polypeptide abnormalities in cells, reducing the need for multiple tests and improving the accuracy of disease diagnosis and treatment planning.
Implementation Method 1
labeling and amplifying, within the particle, a target nucleic acid in the particle, labeling a target polypeptide and other target nucleic acid different from the first target nucleic acid of the particle, and measuring the labeled target nucleic acid and the labeled target polypeptide and other labeled target nucleic acid
Implementation Method 2
measuring the labeled target nucleic acid and the labeled target polypeptide and other labeled target nucleic acid
Data Source
AI summary
A particle measuring method, a sample processing method, and a particle imaging apparatus capable of efficiently acquiring information effective in pathological diagnosis or the like are provided. The particle measuring method comprises a step (S13) of labeling and amplifying, within a particle, a target nucleic acid in the particle, a step (S14) of labeling a surface target polypeptide on the surface of the particle and/or target nucleic acid different from the first target nucleic acid, and a step (S2) of measuring the labeled target nucleic acid and the labeled target polypeptide and/or other labeled target nucleic acid.