Combination marker used for liver cancer detection and application thereof
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A marker, liver cancer technology, applied in the field of biomedicine, can solve only 33% of the problems
Active Publication Date: 2020-03-24
CANCER INST & HOSPITAL CHINESE ACADEMY OF MEDICAL SCI +1
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For nodules larger than 20mm in ultrasonography, the diagnostic accuracy rate is still
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Embodiment 1
[0061] Embodiment 1, the establishment of the method for liver cancer detection
[0062] 1. Obtain markers for liver cancer detection
[0063] The inventors of the present invention combined various databases and comprehensive clinical information, designed probes to enrich the methylation sites of related genes through large-scale screening, and then compared the cfDNA 3-6 months before the diagnosis of liver cancer by building a library and sequencing The difference in methylation level between samples and cfDNA samples of non-liver cancer patients was used to obtain markers for liver cancer detection. The marker consists of TBX15 (GeneID: 6913), AK055957 (GenBank: AK055957.1), BDH1 (GeneID: 622), C6orf223 (GeneID: 221416), DAB2IP (GeneID: 153090), PPFIA1 (GeneID: 8500), LRRC4 ( GeneID: 64101), NFIX (GeneID: 4784), PSD4 (GeneID: 23550), GRASP (GeneID: 160622), OPLAH (GeneID: 26873) and F12 (GeneID: 2161).
[0064] According to the nucleotide sequences of the above-mentione...
Embodiment 2
[0099] Embodiment 2, accuracy experiment
[0100] Select 11 cases of liver cancer tissue samples and 12 cases of white blood cell samples from healthy people, and measure their HCCscore. The names and types of the samples are listed in columns 1 and 2 in Table 7. For each sample to be tested, the following experiments were performed:
[0101] 1. Same as 1 in step 2 of Example 1.
[0102] 2. Use The DNA Mini Kit extracts the genomic DNA of the sample to be tested, and obtains the genomic DNA of the sample to be tested (OD 260 / 280 between 1.8-2.1).
[0103] 3. Take the genomic DNA of the sample to be tested and use EZ DNA Methylation-Lightning TM The Kit performs bisulfite conversion and then purification to obtain the converted DNA of the sample to be tested.
[0104] 4. Replace the "cfDNA transformed by the test subject" in step 2 of Example 1 with the transformed DNA of the test sample, and the other steps remain unchanged.
[0105] 5. Same as 5 in Step 2 of Exampl...
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Abstract
The invention discloses a combination marker used for liver cancer detection and application thereof. The combination marker used for liver cancer detection comprises the respective methylation levelsof adenylate kinase 055957 (AK055957), 3-hydroxybutyrate dehydrogenase 1 (BDH1), chromosome 6 open reading frame 223 (C6orf223), disabled homolog 2-interacting protein (DAB2IP), human coagulation factor XII (F12), Golgi reassembly stacking protein (GRASP), leucine-rich repeat-containing protein 4 (LRRC4), nuclear factor I/X (NFIX), 5-oxoprolinase (ATP-hydrolysing) (OPLAH), recombinant protein tyrosine phosphatase F interacting protein 1 (PPFIA1), Pleckstrin homology and SEC7 domain-containing protein 4 (PSD4) and T-box 15 (TBX15). Experiments prove that whether the genome DNA or plasma circulating free DNA (cfDNA) of to-be-detected liver tissue comes from patients with liver cancer or patients without liver cancer can be judged by detecting the combination marker, and accuracy is relatively high; and the combination marker has significant application value.
Description
technical field [0001] The invention belongs to the field of biomedicine, in particular to a combined marker for detecting liver cancer and its application. Background technique [0002] At present, the diagnosis and treatment of liver cancer still faces major problems, such as difficulty in early diagnosis, high rate of recurrence and metastasis, resistance to radiotherapy and chemotherapy, and lack of effective therapeutic targets. In fact, for any type of cancer, early screening is always a key step in the fight against cancer. [0003] Routine monitoring and screening indicators for early screening of liver cancer mainly include alpha-fetoprotein (AFP) and liver ultrasonography (US). Based on reports and clinical practice at home and abroad, the diagnostic sensitivity of AFP is about 60%, the misdiagnosis rate is 40%, the specificity is about 80%, and the misdiagnosis rate is 20%. AFP is an important reason for being used as a screening index. Ultrasonography has a di...
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