Multi-PCR method and kit for detecting human RhD blood type and gene type

Abstract

The invention relates to a multi-PCR method capable of detecting a human RhD blood type and gene type and an in-vitro diagnosis kit prepared using same, which can detect the human RhD blood type and gene type comprising gene types of D/D, D/d, DEL, DEL/d and d/d. The method is finished by utilizing the change of the specific sequence sites of different gene types of the human RhD blood type, designing a plurality of pairs of primers and carrying out a polymerase chain reaction (PCR). Thereby, the invention can be regarded as being established on the basis of the principle of PCR-SSP (sequence specific primer). By utilizing the sequence specific primer designed aiming at different sites and optimizing the mixing proportion, the reaction buffer components and the PCR reaction conditions of the sequence specific primer, the purpose of accurately detecting the human RhD blood type and gene type is achieved. In one implementation, the invention provides the multi-PCR method for detecting the human RhD blood type and gene type; and in another implementation, the invention provides the kit for detecting the human RhD blood type and gene type. The details of one or a plurality of implementations of the invention are described in the accompanying drawing and the specification. By reading the appended drawings, the detailed descriptions and the claims, a user can clearly learn about other characteristics, purposes and advantages of the invention.

Description

technical field [0001] The invention relates to a multiplex PCR method capable of detecting human RhD blood type genotypes, and an in vitro diagnostic kit prepared by applying the method, which can detect human RhD blood type genotypes, including D / D, D / d, DEL, DEL / d and d / d genotypes. technical background [0002] In this paper, the term "Rh" is the first two letters of the foreign language name of the rhesus monkey (Rhesus Macacus). Since the RhD antigen was discovered in 1939, it has become an important antigen in red blood cells after ABO blood type because it causes hemolytic disease of the newborn and extravascular delayed hemolytic transfusion reaction. The antigenicity of the Rh blood group is very strong, especially the D antigen is the strongest, second only to the A and B antigens of the ABO system. Those who have RhD antigen on human red blood cells are Rh positive, otherwise they are negative. The RhD-negative blood type accounts for about 3 to 5‰ of Chinese...

AI Technical Summary

Problems solved by technology

[0007] 1. Expensive: Although the D gene detection reagents produced by Germany’s BAG and INNO-TRAIN companies are reliable, they cost about 1200-1300 RMB per person, which is very expensive; A copy costs about 100 RMB, but the reliability is poor, and this price is still unaffordable for large-scale testing;

[0008] 2. The operation is cumbersome: the D gene detection reagents of Germany’s BAG and IVIN companies need 4 to 8 reaction tubes per person, while the American G&T reagents need 12 reaction tubes per person, and the consumption of sample DNA and consumables is very large , and the addition of samples is cumbersome, error-prone, and difficult to interpret, so it is not suitable for large-scale testing;

[0009] 3. The internal control is the human growth hormone gene, but in our actual use, we found that there are amplification bands in the internal control, but there is no amplification product in the detection band. It is impossible to judge whether some negative results are caused by the degradation of the Rh gene.

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