PCR primer and kit for measuring individual RHD gene zygosity and measuring method thereof

Abstract

The invention relates to a PCR primer and a kit for measuring individual RHD gene zygosity and a measuring method thereof. The primer of the invention is used for detecting a first exon of an RHD gene, a 3' end position of an upstream primer is at a -554 basic group opposite to a first basic group position of an RHD gene code area, and a 3' end position of an downstream primer is at a 18th basic group of a first intron of the RHD gene. The kit and the measuring method of the invention based on the specific oligonucleotide primer has sophistication, specificity and high accuracy rate, and has simple and easy operation in the process of measuring.

Description

[0001] This application is a divisional application of the following invention application: the application number of the original application is 200410032010.X, the application date is March 25, 2004, and the title of the invention is "PCR primers and kits and measurement methods for individual RHD gene zygote determination " technical field [0002] The present invention relates to PCR primers for individual RHD gene zygote type determination or RHD gene number determination, kits for individual RHD gene zygote type determination or PHD gene number determination, and individual RHD gene zygote type determination or RHD gene number determination Methods. Background technique [0003] In 2000, Wagner et al. found that there is a highly homologous sequence on both sides of the PHD gene, about 9000 bp, called the upstream Rh box at the 5'-end of the RHD gene, and the downstream Rh box at the 3'-end. The RHD gene deletion of the RHD-negative haploid occurs between the upstream...

AI Technical Summary

Problems solved by technology

In the past, the determination of the number of RHD genes or the RHD zygote type was mainly based on the estimation of the Rh small factor phenotype, or through complex family investigations, or indirect methods such as identification based on the amount of RhD antigen. In 2000, the first RHD gene zygote type was established internationally. Direct assay technology is the restriction fragment length polymorphism (RFLP) method, which uses a pair of PCR primers to simultaneously amplify the downstream and fusion Rh box, and then uses restriction endonucleases for digestion, and then electrophoresis to analyze the size of the fragments. Three RHD zygote types can be judged by one experiment result, but this method is complicated to operate and takes a long time

In 2001 and 2002, Chinese Hong Kong scholars and Austrian scholars independently reported the number of RHD genes in individuals with known Rh-positive phenotypes using the amplification mutation block detection system and real-time polymerase chain reaction (Real-time PCR) technology. , which is also complex and requires special instruments, and cannot distinguish between RHD(+) / RHD(-) and RHD(-) / RHD(-) zygotic types

In 2003, Shao Chaopeng, one of the inventors of this patent, designed and established a simple polymerase chain reaction (PCR) detection method, which can judge three RHD zygote types at one time, but in the subsequent application, it was found that due to the position of the primer and the reaction area, etc. , resulting in a certain rate of false positives and false negatives

Images