Dna probe for enriching low-frequency dna mutation and its application
A DNA probe, low-frequency technology, applied in the direction of DNA / RNA fragments, recombinant DNA technology, microbial measurement / inspection, etc., can solve the problem of high sample size, limit the application of simultaneous detection of multiple sites, etc., to achieve The effect of high sensitivity and low false positive rate
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Embodiment 1
[0046] In this example, the L858R mutation on exon 21 of the EGFR gene hotspot mutation is taken as an example to analyze the method for capturing and enriching EGFR mutations by special DNA probes of the present invention. The samples used in the experiment were Horizon’s Multiplex I cfDNAReference Standard Set (HD780), which were EGFR gene wild type, with 5% EGFR L858R mutation, with 1% EGFR L858R mutation, and with 0.1% EGFR L858R mutation; 3 cases of lung adenocarcinoma Clinical samples.
[0047] The method for detecting the L858R mutation on exon 21 of the EGFR gene hotspot mutation by using the above method comprises the following steps:
[0048] (1) Separation of peripheral blood samples
[0049] 1. Collect 1 tube (10mL / tube) of peripheral blood from the subject into a Streck tube, gently upside down (to prevent cell rupture) 6-8 times and mix thoroughly, and perform the following treatment within 7 days after blood collection;
[0050] 2. Centrifuge at 1600g for 10 m...
Embodiment 2
[0109] In this example, the T790M mutation on exon 20 of the EGFR gene hotspot mutation is taken as an example to analyze the method for capturing and enriching EGFR mutations by special DNA probes of the present invention. The samples used in the experiment were Horizon’s Multiplex I cfDNAReference Standard Set (HD780), which were EGFR gene wild type, with 5% EGFR T790M mutation, with 1% EGFR T790M mutation, and with 0.1% EGFR T790M mutation; two cases of lung adenocarcinoma Clinical samples.
[0110] The method for detecting the T790M mutation on exon 20 of the EGFR gene hotspot mutation by using the above method comprises the following steps:
[0111] (1) Separation of peripheral blood samples
[0112] 1. Collect 1 tube (10mL / tube) of peripheral blood from the subject into a Streck tube, gently invert it upside down (to prevent cell rupture) 6-8 times and mix thoroughly, and perform the following treatment within 7 days after blood collection;
[0113] 2. Centrifuge at 16...
Embodiment 3
[0173] In this example, the deletion mutation on exon 19 of the EGFR gene hotspot mutation is taken as an example to analyze the method for capturing and enriching EGFR mutation with a special DNA probe of the present invention. The samples used in the experiment were Horizon's Multiplex I cfDNAReference Standard Set (HD780), which were EGFR gene wild type, with 5% EGFR 19DEL mutation, with 1% EGFR 19DEL mutation, and with 0.1% EGFR 19DEL mutation; two cases of lung adenocarcinoma Clinical samples.
[0174] The method for detecting the deletion mutation on exon 20 of EGFR gene hotspot mutation by using the above method comprises the following steps:
[0175] (1) Separation of peripheral blood samples
[0176] 1. Collect 1 tube (10mL / tube) of peripheral blood from the subject into a Streck tube, gently invert it upside down (to prevent cell rupture) 6-8 times and mix thoroughly, and perform the following treatment within 7 days after blood collection;
[0177] 2. Centrifuge a...
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