Soluble microneedle patch and preparation method thereof

Abstract

The invention discloses a soluble microneedle patch and a preparation method thereof. The invention provides a whole set of probes which are composed of single-stranded DNA molecules as shown in sequence 1-9 in a sequence table. The invention also discloses a gene chip prepared by immobilizing the whole set of probes on a chip medium. The gene chip can be used for detecting the mutation of the HBsAg sites in to-be-detected samples. During detection, the DNAs of viruses are extracted from the serum samples of to-be-detected patients and then subjected to in-vitro amplification and hybridizationwith the probes in the chip, and the types of mutation can be determined according to hybridization signals. A detection method provided by the invention has the advantages of high throughput, high specificity, simple operation, automatic judgment of results and easy realization of standard control, is applicable to clinical detection of HBsAg gene mutation, and can meet the needs of physical examination and blood donor screening.

Description

technical field [0001] The invention relates to a dissolvable microneedle patch and a preparation method thereof. Background technique [0002] Hepatitis B is caused by infection with the hepatitis B virus (HBV). HBV belongs to the hepadnaviridae family and is a circular double-stranded DNA virus. The outer layer is a lipoprotein envelope containing hepatitis B surface antigen (HBsAg); the inner layer is a core containing nucleic acid and hepatitis B core antigen. [0003] The HBV S gene encodes surface antigen (HBsAg). HBsAg has important biological significance, is the main component of hepatitis B vaccine, and is an important basis for diagnosing hepatitis B virus infection. The S gene region can be divided into three segments: the S region, the pre-S1 region and the pre-S2 region. Each of the three segments has a start codon (AUG) at the 5' end, but shares a stop codon. The translated products form the major protein (226 amino acids), medium protein (281 amino acids) ...

AI Technical Summary

Problems solved by technology

Mutations in the S gene region, especially the mutation at the amino acid 145 position of the "α" determinant, will lead to changes in the antigenicity of HBsAg, making it undetectable by the widely used HBsAg diagnostic reagents in clinical practice, resulting in missed detection, which may lead to Cause such virus carriers to become blood donors, bringing serious consequences to blood recipients

[0004] At present, there are four main molecular detection methods for HBsAg gene variation: (1) sequencing method: the gold standard for HBV gene detection, with high accuracy, but low sensitivity and long time-consuming; (2) RFLP method: the principle is to The DNA fragment to be tested is digested with a restriction endonuclease to identify and cut the specific sequence, and the digested product is subjected to electrophoresis to determine whether the target gene has a specific sequence according to the size of the DNA fragment, but the degree of standardization Low, poor ability to detect mixed genotypes. When detecting gene mutations, multiple detection systems are required to detect mutations of more than 1 base; (3) Quantitative PCR method: high sensitivity, but low throughput, It is difficult to detect multi-gene locus variation; (4) Reverse linear probe hybridization method: first point known probes on nitrocellulose membrane or nylon membrane respectively, and PCR amplification with 5' end-labeled biotin primers The product is hybridized, and then the hybridization signal is displayed by the corresponding color reaction

The naked eye interpretation is easy to misjudge, the detection cost is high, and it is difficult to carry out routinely in domestic clinical laboratories

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