Primer combination for detecting mutation of PIK3CA gene in trace tissue and application of primer combination

Abstract

The invention discloses a primer combination for detecting mutation of a PIK3CA gene in trace tissue. The primer combination comprises a pre-amplification primer group, a primer group for detecting No.9 exon mutation and a primer group for detecting No.20 exon mutation. According to a method, a trace amount of DNA is pre-amplified to obtain a high-concentration DNA template, and the mutation condition of the PIK3CA gene in a sample is detected by combining a direct sequencing method; by applying the primer combination to preparation of a detection reagent for detecting mutation of the PIK3CA gene, the DNA concentration of the detectable sample is low up to 100 pg, and all mutation of a No.9 exon and a No.20 exon of the PIK3CA gene can be simultaneously detected. The detection method is high in sensitivity and specificity, low in cost and suitable for PIK3CA gene mutation detection on a clinical tumor patient and has the very good clinical application value.

Description

technical field [0001] The invention belongs to the technical field of biological detection, in particular to a PIK3CA Primer combination for gene mutation detection and its application in tumor drug selection and disease diagnosis. Background technique [0002] Phosphatidylinositol-3-kinase (PI3K) is a phosphatidylinositol kinase that can phosphorylate the third hydroxyl group of the inositol ring, and has dual activities of phosphatidylinositol kinase and serine-threonine protein kinase (Fruman , D. A. et al.2014), regulating various biological functions such as cell proliferation, survival, migration, apoptosis and cell cycle. The PIK3CA gene encodes the class IA PI3K catalytic subunit P110α, which is located at 3q26.3 and contains 21 exons (Volinia, S. et al. 1994). KANG S et al found that, PIK3CA Gene plays an oncogene role in a variety of tumors, present in about 30% of human solid tumors PIK3CA Gene mutations, 80%-90% of which occur in the 9th and 20th exons of th...

AI Technical Summary

Problems solved by technology

However, the application of the last base mismatch in ARMS technology cannot completely block the amplification of wild-type DNA, there is a risk of false positives, and it can only be detected for specific sites

In addition, such as high performance liquid chromatography, capillary electrophoresis, etc., require special equipment, and the operation is complicated, which is not conducive to large-scale clinical promotion

Nucleic acid sequence amplification (NASBA), self-sequencing sequence replication (3SR) and strand displacement replication (SDA) are all isothermal amplification methods, but their specificity for the amplification of target sequences is not strong

[0005] despite this, PIK3CA The detection method of mutation is still based on the detection of tumor tissue samples as the gold standard, but in clinical practice, the detection is often limited due to insufficient tissue samples

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