Primer group and detection method for detecting pancreatic cancer
A primer set and pancreatic cancer technology, applied in the field of tumor genes, can solve the problems that the detection standards of pancreatic cancer CTCs have not been established, and achieve the effects of fast detection speed, simple operation, and low cost
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Embodiment 1
[0033] Example 1: Gene selection
[0034] Many studies have shown that the sensitivity of single-gene screening is about 20-60%. Most of the single-gene tests used for early tumor screening have low detection sensitivity or specificity, which cannot meet clinical needs. Multi-gene combined detection can effectively solve the problem of low sensitivity and improve the accuracy of early tumor screening and diagnosis. Zhou J et al. (Zhou J, Hu L, YuZ, et al. Marker expression inducing cancer cells of pancreatic cancer patients. J Surg Res 2011; 171:631–6.) simultaneously detected hTERT, CK20, C-met, and CEA genes, and the results It shows that the sensitivity of combined detection of four genes for early diagnosis of pancreatic cancer is much higher than that of single gene detection. In order to increase the detection rate of early tumors, improve the cure rate of tumor patients, and improve the prognosis of patients, we screened the differentially expressed genes between pancr...
Embodiment 2
[0035] Embodiment 2: primer screening
[0036](1) Design primers a1~a3, b1~b3, c1~c3, d1~d3, e1~e3, f1~f3, g1~g3, h1~h3, i1~i3, specifically: use bioinformatics software to The A-I sequence of the target gene was analyzed, and a specific primer set was designed by using the sequence analysis software, and the specificity of each paired primer in the human genome was detected by the NCBI primer search software, and three pairs of specific amplification primers a1~a3, b1~ b3, c1~c3, d1~d3, e1~e3, f1~f3, g1~g3, h1~h3, i1~i3;
[0037] Table 1: PCR Detection Primers
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[0042] (2) Processing of peripheral blood samples
[0043] The peripheral blood of pancreatic cancer patients admitted to a hospital and confirmed by pathology was collected. Fresh blood was collected through the cubital vein at 7:00 am on an empty stomach, stored in an anticoagulant tube, shaken well, and stored at room temperature for ≤4hr.
[0044] 2.1 Add an equal...
Embodiment 3
[0075] Example 3: Effect Verification
[0076] According to the screening results in Example 2, 200 tumor samples were tested using the primer sets described in the table below.
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[0079] Among them, the forward primer of CK8 is CGCCTGGAAGGGCTGACCGA, the reverse primer is GTTGGCAATATCCTCGTACT; the forward primer of N-cadherin is CCTTAACTGAGGAGTCAGTG, and the reverse primer is CAGACCTGATCCTGACAAGC; the forward primer of VEGF is GTGCCCGCTGCTGTCTAATG, and the reverse primer is CACATCTGCAAGTACGTTCG; The primer was GAGAAGCAGAGCCATGTGGT and the reverse primer was GCACTCTTCCTCCAACTGCC.
[0080] The detection rates of primer sets 1 to 9 are shown in the above table. It can be seen from the table that, in the primer set, as the number of primers increases, the detection rate can be improved to a certain extent. Further, by comparing the primer sets No. 5 to No. 9, it can be found that the combination in the primer set has an important influence on the detection ...
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