Tumor cell microsatellite instable state detection system
A technology for microsatellite instability and tumor cells, applied in the fields of multiplex fluorescent PCR technology and capillary electrophoresis technology, can solve the problems of false positive or false negative, long detection cycle, complicated operation, etc.
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[0099] Example 1:
[0100] A 9-site MSI detection system was used to detect colorectal cancer tumor tissue sample A and its corresponding normal tissue samples.
[0101] 1.1 Detection system
[0102] The detection kit includes PCR reaction solution, primer mixture, enzyme mixture, positive control, negative control, internal standard, etc. The primer mixture includes 9 pairs of primers with 6 single nucleotide repeat sites and 3 control sites. The primer sequences are as follows:
[0103] NR-21 primer:
[0104] Forward primer: 5’-GAGTCGCTGGCACAGTTCTA-3’
[0105] Reverse primer: 5’-FAM-ATATTCCTACTCCGCATTCACAC-3’
[0106] NR-24 primer:
[0107] Forward primer: 5’-TTGCTGAATTTTACCTCCTGAC-3’
[0108] Reverse primer: 5’-TAMAR-ATTGTGCCATTGCATTCCAA-3’
[0109] NR-27 primer:
[0110] Forward primer: 5’-GGAAACAAAGCATTGAAGTCTGCAGT-3’
[0111] Reverse primer: 5’-HEX-GAGGTTCTGAGTCGATAATACTAGC-3’
[0112] BAT-25 primer:
[0113] Forward primer: 5’-CTCGCCTCCAAGAATGTAAGT-3’
[0114] Reverse primer: 5’-HEX-TCTGC...
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[0150] Example 2:
[0151] The 12-site MSI detection system was used to detect colorectal cancer tumor tissue samples B and C and their corresponding normal tissue samples.
[0152] 2.1 Detection system
[0153] The detection kit includes PCR reaction solution, primer mixture, enzyme mixture, positive control, negative control, internal standard, etc. There are 12 pairs of primers used in the examples, and the primer sequences are as follows:
[0154] The sequences of NR-21 primer, NR-24 primer, NR-27 primer, BAT-25 primer, BAT-26 primer, MONO-27 primer, PentaC primer, PentaD primer, and Amel primer are the same as in Example 1.
[0155] Braf V600E primers:
[0156] Forward wild-type primer: 5’-GGTGATTTTG GTCTAGCTAC A T-3’
[0157] Forward mutant primer: 5’- GGTT GGTGATTTTGGTCTAGCTAC GA-3’
[0158] Reverse common primer: 5’-TAMAR-GTTGAGACCTTCAATGACTTTCTA-3’
[0159] UGT1A1*6 sites:
[0160] Forward wild-type primer: 5’-CCTCGTTGTA CATCAGAG CG-3’
[0161] Forward mutant primer: 5’- CCTT ...
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