Group of probes, detection kit and detection method for detecting thalassemia gene point mutation based on liquid chip of locked nucleic acid sensibilization

Abstract

The invention discloses a group of probes, a detection kit and a detection method for detecting thalassemia gene point mutation based on a liquid chip of locked nucleic acid sensibilization. Due to the characteristics that locked nucleic acid can flexibly modify probes, probes for detecting thalassemia gene point mutation are designed; these probes are respectively coupled to different numbers of microspheres; a specific primer disclosed by the invention is designed to amplify mutation sites, so as to obtain a PCR product; the PCR product is hybridized with the microspheres coupled to the probes; fluorescence labeling is carried out by using a fluorescence labeling reagent; and finally detection on the mutation sites is carried out through a liquid chip detector. The probes based on LNA sensibilization are shortened in comparison with a conventional probe, and still can keep relatively high Tm values; so that the hybridization signal ratio of completely matched probes and mismatched probes is improved to over four times from below twice in the past, thus the probes are easy to distinguish.

Description

Technical field: [0001] The invention belongs to the field of molecular diagnosis, and in particular relates to a group of probes, detection kits and detection methods for detecting thalassemia gene point mutations based on locked nucleic acid-sensitized liquid-phase chips. Background technique: [0002] Thalassemia (Thalassemia) is a genetic hemolytic disease caused by globin synthesis disorders, divided into two types: α-thalassemia (abbreviated as α-thalassemia) and β-thalassemia (β-thalassemia). The disease is mainly found in countries along the Mediterranean Sea and Southeast Asian countries. It is the single gene disease with the widest distribution and the largest cumulative population in the world. There are about 90 million thalassemia gene mutation carriers in the world. It is more common in the south of my country, and it is one of the genetic diseases with the highest incidence and the greatest impact in the provinces south of the Yangtze River, especially in Gua...

AI Technical Summary

Problems solved by technology

If there is only one base difference between wild-type and mutant DNA, probes designed according to conventional methods will often produce high non-specific hybridization signals, resulting in the inability to clearly distinguish wild-type and mutant DNA fragments

Images