Lung Biopsy Sample Preparation for Nucleic Acid Preservation
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Solution Overview
Problem
The detection of multiple gene mutations in lung cancer specimens is hindered by the degradation of nucleic acids caused by formalin fixation and paraffin embedding, and the limited availability of neoplastic nucleic acids in cytologic specimens and plasma cell-free DNAs, leading to suboptimal sample quality for next-generation sequencing.
Innovation Solution
A method involving the use of negative pressure to release cells from lung biopsy tissues into an isotonic solution, mixing with cytologic specimens to create a cell suspension, and treating with ammonium sulfate to extract and stabilize nucleic acids, thereby bypassing formalin fixation and paraffin embedding, which degrades nucleic acids and improves nucleic acid yield and amplification.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If formalin fixation and paraffin embedding are used to preserve lung cancer specimens, then tissue structure is maintained for pathological diagnosis, but nucleic acid degradation occurs reducing sample quality for genetic analysis
Solution Approach 1:
The invention separates the specimen into two distinct pathways: one for tissue structure preservation (formalin fixation and paraffin embedding for pathological diagnosis) and another for nucleic acid preservation (fresh frozen specimens for genetic analysis). This segmentation allows both requirements to be met simultaneously by using different preservation methods for different analytical purposes.
Solution Approach 2:
The invention introduces an intermediary step of collecting cytologic specimens (bronchoalveolar lavage fluid, bronchial washing fluid, or puncture needle washing fluid) that contain neoplastic cells. These cytologic specimens serve as a bridge, providing accessible nucleic acid sources that can be processed separately without compromising the formalin-fixed tissue blocks needed for pathological diagnosis.
2Adaptability or versatility
If multiple gene mutation detection by next-generation sequencing is performed, then comprehensive molecular profiling is achieved, but the limited amount of neoplastic nucleic acids in cytologic specimens becomes insufficient
Solution Approach 1:
The invention merges multiple specimen sources to accumulate sufficient neoplastic nucleic acids. It combines cytologic specimens (bronchoalveolar lavage fluid, bronchial washing fluid, or puncture needle washing fluid) with fresh frozen tissue specimens, thereby pooling the nucleic acid content from multiple sources to achieve the required quantity for comprehensive multiple gene mutation detection by next-generation sequencing.
Solution Approach 2:
The invention performs preliminary enrichment of neoplastic cells from cytologic specimens before nucleic acid extraction. By pre-concentrating the neoplastic cells through cytological examination and selection, sufficient quantities of nucleic acids are obtained in advance, enabling subsequent multiple gene mutation detection without being limited by the initial scarce nucleic acid amount.
3Duration of action of stationary object
If outdated specimens are used for next-generation sequencing, then sample availability is maintained, but nucleic acid degradation increases affecting detection accuracy
Solution Approach 1:
The invention changes the storage parameters from formalin fixation (chemical preservation) to fresh frozen storage (physical preservation at low temperature). This parameter change maintains nucleic acid integrity over extended storage periods while preserving the ability to perform sensitive next-generation sequencing analysis, thereby resolving the contradiction between long-term storage and detection accuracy.
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 produces a sample with less degradation and higher nucleic acid yield, suitable for genetic analysis by next-generation sequencing, enhancing the determination of molecular target drugs and improving the precision of lung cancer treatment regimens.
Implementation Method 1
treating a biopsy tissue taken from a lung cancer patient with negative pressure in an isotonic solution to release a cell from the biopsy tissue
Implementation Method 2
treating with ammonium sulfate to extract and stabilize nucleic acids
Data Source
AI summary
A method for preparing a sample from a specimen includes treating a biopsy tissue taken from a lung cancer patient with negative pressure in an isotonic solution to release a cell from the biopsy tissue, collecting the isotonic solution containing the released cell, and mixing the collected isotonic solution and a cytologic specimen collected from the lung cancer patient together to obtain a cell suspension, wherein the cytologic specimen is one or more selected from the group consisting of a bronchial lavage fluid, a bronchoalveolar lavage fluid, a bronchial scraping lavage fluid and a puncture needle lavage fluid, and wherein the biopsy tissue and the cytologic specimen contain a tumor cell.