Real-time fluorescent PCR (polymerase chain reaction) detection kit for non-deletion type alpha thalassemia

Abstract

The invention relates to the field of a biological medicine, and particularly relates to a real-time fluorescent PCR (polymerase chain reaction) detection kit for non-deletion type alpha thalassemia. The kit comprises two PCR reagents, namely a PCR reagent I and CPR reagent II, wherein each PCR reagent comprises a primer, a probe, PCR Buffer, dNTPs, MgCl2, DNA (deoxyribonucleic acid) polymerase, UNG (unguentum) enzyme and glycine betaine. By adopting the kit, QS (quality safety) and CS mutant types of the non-deletion type alpha-thalassemia can be simultaneously detected by a single test. The real-time fluorescent PCR detection kit has the characteristics of short detection time, high sensitivity and strong specificity, and is simple to operate; the result can be judged and read after fluorescent PCR is finished; the problem of carrying out complicated operations of hybridization, sequencing and solubility curve analysis on a PCR product in the prior art is overcome.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a real-time fluorescent PCR detection kit for non-deletion type α-thalassemia. Background technique [0002] Thalassemia, also known as thalassemia, is an inherited hemolytic anemia. Its pathogenesis is the reduction or loss of globin chains in the synthesis of hemoglobin, resulting in abnormal structure of hemoglobin. Thalassemias are classified according to the amino acid chains involved, and thalassemias are usually divided into 4 types: α, β, δβ, and δ, among which β and α thalassemias are more common. [0003] In foreign countries, thalassemia is more common in Mediterranean countries (hence the name thalassemia) and Southeast Asian countries. The vast area south of the Yangtze River in my country is a high incidence area of ​​thalassemia, especially in the three provinces (regions) of Guangxi, Guangdong, and Hainan, and other provinces (regions) such as Hong Kong, Taiwan, Fujia...

AI Technical Summary

Problems solved by technology

These two methods involve many operational steps, and the results cannot be directly interpreted after PCR. Subsequent analysis of the PCR products is required. The operation of opening the tube is prone to contamination, and the detection process takes a long time.

[0014] However, when using the TaqMan probe fluorescent PCR method, the design is relatively difficult, especially the design of the TaqMan probe, which must not only ensure specificity but also satisfy effective and efficient amplification, and at the same time need to solve the gap between channels in single-tube multi-channel detection. The problem of signal interference, as well as the problem that there were not many users of fluorescent PCR instruments before, have affected the application of TaqMan fluorescent PCR method.

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