Multiplex polymerase chain reaction (PCR) method and kit for detecting human RhD blood type genotypes

Abstract

The invention relates to a multiplex polymerase chain reaction (PCR) method capable of detecting human RhD blood type genotypes and an in vitro diagnostic kit prepared by applying the method and capable of detecting the human RhD blood type genotypes comprising D / D, D / d, DEL, DEL / d and d / d genotype. According to the method, variation of specific sequence loci of the different genotypes of a human RhD blood type is utilized to design a plurality of pairs of primers to perform a PCR. Accordingly, the multiplex PCR method and the in vitro diagnostic kit can be regarded to be on the basis of a PCR-SSP (sequence specific primer) principle. By designing the sequence specific primers according to different loci and by optimizing mixing proportion, reaction buffer solution ingredients and PCR reaction conditions of the sequence specific primers, the aim of accurately detecting the human RhD blood type genotypes is achieved. In an embodiment, the multiplex PCR method for detecting the human RhD blood type genotypes is provided, and in another embodiment, the kit for detecting the human RhD blood type genotypes is provided. Details of one or a plurality of embodiments are described in attached drawings and explanations. By reading the attached drawings, detailed descriptions and claims, other characteristics, aims and advantages of the multiplex PCR method and the kit can be learned about clearly.

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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