Method and probe for detecting cis-trans mutation configurations of EGFR-T790M and EGFR-C797S
A technology of EGFR-C797S and EGFR-T790M, which is applied in biochemical equipment and methods, measurement/testing of microorganisms, DNA/RNA fragments, etc., can solve the problems of long time consumption, drug resistance, high detection sensitivity, etc., and achieve specificity Effects of High, Reduced Sensitivity, and High Sensitivity
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Embodiment 1
[0139] This embodiment provides a kit for detecting cis-trans mutation types of EGFR-T790M and EGFR-C797S, the composition of which is shown in the following table:
[0140] Table 2
[0141]
[0142]
[0143] (2) The experimental procedure of this kit is as follows
[0144] table 3
[0145]
[0146] (1) Comparison of blocking performance of Blocker 1:
[0147] Use plasmid DNA with different mutation information as a template, and the plasmid DNA information is shown in the table below:
[0148] Table 4
[0149] name mutation type Plasmid 001 wild type (WT) Plasmid 002 T790M C>T Plasmid 003 C797S T>A Plasmid 004 C797S G>C Plasmid 005 T790M C>T&C797S T>A Plasmid 006 T790M C>T&C797S G>C
[0150] The system was prepared according to the formula in the table below, and the PCR program was shown in Table 3.
[0151] table 5
[0152]
[0153]
[0154] The result is as Figure 3-8 As shown, Blocker 1 block...
Embodiment 2
[0196] This embodiment provides a kit for detecting cis-trans mutation types of EGFR-T790M and EGFR-C797S. The difference from the embodiment is that the following probes are used:
[0197] Table 11
[0198]
[0199] Use plasmid DNA with different mutation information as a template, and the plasmid DNA information is shown in the table below:
[0200] Table 12
[0201] name mutation type Plasmid 001 wild type (WT) Plasmid 002 T790M C>T Plasmid 003 C797S T>A Plasmid 004 C797S G>C
[0202] The system was prepared according to the formula in the table below, and the PCR program was shown in Table 3.
[0203] Table 13
[0204]
[0205] The product is sent for inspection and next-generation sequencing: the results are as follows
[0206] Plasmid 001 (wild type) Blocker 3 product sequencing results are as follows Figure 55 Shown; Blocker 4 product sequencing results are as follows Figure 56 Shown: Blocker 3 product has T790M C>...
Embodiment 3
[0212] This embodiment provides a kit for detecting cis-trans mutation types of EGFR-T790M and EGFR-C797S. The difference from the embodiment is that the following probes are used:
[0213] Table 14
[0214]
[0215] The system was prepared according to the formula in the following table, the DNA samples are shown in Table 4, and the PCR program is shown in Table 3.
[0216] Table 15
[0217]
[0218] Blocker5 amplification results are as follows Figures 63-68 Shown: Blocker 5 blocks wild-type and single-point C797S mutation DNA amplification, does not block or less blocks single-point T790M mutation and cis mutation T790M C>T&C797S T>A or G>C DNA amplification.
[0219] Blocker6 amplification results are as follows Figures 69-74 As shown, Blocker 6 blocks wild-type and single-point T790M mutant DNA amplification, does not block or less blocks single-point C797S T>A or G>C and cis-mutant T790M C>T&C797S T>A or G>C DNA amplification.
[0220] Comparison of blocking...
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