Detection kits for detecting HLA-B27 gene and method

Abstract

The invention discloses a group of detection kits for rapidly detecting an HLA-B27 gene and establishes a relatively efficient and sensitive method for rapidly detecting the HLA-B27 gene. The detection kits for rapidly detecting the HLA-B27 gene are convenient to use, simple to operate and good in detection effect, greatly simplifies the operation process and reduce pollution during operation, have the characteristics of high sensitivity, high specificity and high selectivity and realize typing detection of the HLA-B27 gene. The provided detection method adopts completely closed tube operation and is simple, convenient and quick to operate; detection results are obtained by directly detecting a fluorescence signal Ct value in a PCR direct detection process, post-processing such PCR electrophoresis detection is not needed, the defects that pollution is easily caused and the false positive rate is high with the conventional PCR technology are overcome, and the method can effectively avoid non-specific amplification and is suitable for large-volume sample detection.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a detection kit and a method for detecting HLA-B27 gene by fluorescent quantitative PCR technology. Background technique [0002] Human leukocyte antigen (HLA), also known as major histocompatibility antigen (MHC), is located in the 6p21.3 region of the short arm of chromosome 6, with a total length of about 3.6Mb. The HLA gene is the most complex human gene system known so far. According to its arrangement on the chromosome, it can be divided into three categories: class I gene region includes HLA-A, HLA-B and HLA-C; class II gene region includes HLA- DR, HLA-DP and HLA-DQ; class III gene regions are located between the above two types of gene regions, and mainly encode products such as complement and heat shock proteins. [0003] In 1973, Brewerton first reported that human histocompatibility antigen HLA-B27 (human leucocyteantigen B27, HLA-B27) was highly correlated w...

AI Technical Summary

Problems solved by technology

However, due to the particularity of the gene structure and the designed PCR primers and other factors, there may be overlapping peaks in the sequencing results, which is not conducive to the interpretation of the results

Moreover, sequencing has special requirements for reagents and instruments, complicated operation, relatively high cost, slow speed, and low throughput, and is not suitable for rapid auxiliary diagnosis of clinical diseases

The principle of the PCR-SSP method is to design specific primers for direct PCR amplification based on known HLA-B27 gene fragments, and detect the PCR products by agarose gel electrophoresis. The results are intuitive and easy to promote in grassroots medical and health units. However, the biggest drawback of this method is that there are many steps, the operation is easy to pollute, and it is prone to false positives. It cannot be batched and automated.

Flow cytometry (FCM) has the advantages of rapidity, high degree of automation, and is suitable for large-scale sample detection, but the specificity of the fluorescent monoclonal antibody relied on in this method is not high, and it is easy to mix with many other HLA antigens, such as HLA-B7 , HLA-B22, etc. have interference reactions, which affect the accuracy of the results

In addition, flow cytometry is a serological level test and cannot determine the exact genotype of the sample

Moreover, flow cytometry equipment is expensive in materials, high in maintenance costs, and has high technical requirements for operators, which limits its promotion and application in primary medical units.

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