Cervical CTC examination refers to a CTC examination of the cervix, mainly to determine whether there are tumor cells in the cervix. CTC examination is widely used in clinical medicine, especially in oncology, where it is a very necessary examination method. There are many different methods and detection methods for CTC examination. Below, this article will give a relatively detailed introduction to the specific items of cervical CTC examination and monitoring methods.
CTC is the abbreviation of Circulating Tumor Cell, which means circulating tumor cells. It is a general term for various tumor cells present in peripheral blood. CTC detection captures and detects CTCs in trace amounts in peripheral blood and monitors the trend of changes in CTC types and quantities, so as to monitor tumor dynamics in real time, evaluate treatment effects, and achieve real-time individual treatment. 2. Methods of CTC Detection CTC testing is considered a "liquid biopsy" because it only requires 5-10 ml of blood and is convenient, non-invasive, and has no side effects. CTC detection is a new non-invasive diagnostic tool that provides important guidance for the prognosis, efficacy evaluation and individualized treatment of cancer patients by accurately counting and typing tumor cells in the peripheral blood circulation system. Among them, the circulating tumor cell separation and typing technology launched by Meiji Bio is at the international leading level, providing technical support for non-invasive real-time monitoring and real-time individualized treatment of tumors.
Commonly used CTC detection techniques include immunofluorescence, PCR, FISH and high-throughput sequencing. 1. Immunofluorescence (IF) method: Solid tumor cells are generally of epithelial origin, and cytokeratin (CK) or other tumor markers (such as HER2) are expressed in the cells. With the help of immunofluorescence staining, tumor markers in CTCs from solid tumors can be identified, thereby achieving the purpose of detecting CTCs. This is also the most common method for detecting CTCs. The disadvantage is that during the mesenchymal transformation (EMT) stage of CTC formation, a large amount of CK will be degraded, resulting in false negatives due to "invisibility" during CTC detection. 2. Fluorescence in situ hybridization (FISH) Fluorescence in situ hybridization is based on known specific DNA sequences in cells. It uses fluorescently labeled specific oligonucleotide fragments as probes to hybridize with DNA molecules in the genome of the cell to detect the presence and abundance of the specific gene sequence. The combination of FISH technology and CTC can not only detect CTC surface markers, but also detect markers and karyotypes inside CTC cells. 3. RT-PCR reverse transcription PCR. Total RNA is extracted from tissues or cells, and the mRNA therein is used as a template. Primers and reverse transcriptase are used to reverse transcribe the mRNA into cDNA. The cDNA is then used as a template for PCR amplification to obtain the target gene or detect gene expression. RT-PCR has increased the sensitivity of RNA detection by several orders of magnitude, making it possible to analyze some extremely trace RNA samples. Combined with RT-PCR, the expression of several genes can be detected simultaneously. Site-specific PCR technology can be used to detect mutations in driver genes carried by CTCs.4. The number of CTCs in next-generation sequencing is small, and it is difficult to perform second-generation sequencing directly. The DNA of a single cell needs to be amplified and then the gene sequence can be checked using next-generation sequencing. The more representative ones are multiple annealing and circularization cycle amplification technology and multiple displacement amplification (MDA). However, the technology of single-cell sequencing is currently only in the laboratory stage, and there is still a long way to go before it can be promoted to the market in the future. Moreover, due to the heterogeneity of tumor cells, more scientific research is needed to prove whether single-cell sequencing is representative. |
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