Unlock instant, AI-driven research and patent intelligence for your innovation.

HLA subtype detection kit and application of kit

A detection kit and kit technology, applied in the field of HLA subtype detection kits, can solve problems that plague clinicians, skin adverse reactions, etc., and achieve the effect of improving sensitivity and specificity

Pending Publication Date: 2020-06-05
THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV
View PDF8 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this class of drugs often causes cutaneous adverse reactions (cADRs), which have long puzzled clinicians

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • HLA subtype detection kit and application of kit
  • HLA subtype detection kit and application of kit
  • HLA subtype detection kit and application of kit

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1. Multiplex amplification 9 kinds of HLA subtype detection kits of the present invention

[0021] 1. Design of primers and probes for detection of 9 HLA subtypes

[0022] This kit is aimed at 9 HLA gene subtypes found in human genomic DNA and related to epilepsy drug allergy: HLA-A*02:01, HLA-A*24:02, HLA-B*15:01, HLA-B *15:02, HLA-B*15:11, HLA-B*35:01, HLA-B*38:02, HLA-B*13:01, and HLA-DRB1*04:06. Specific primers and probes were designed, wherein the specific primers were multiplexed and amplified to obtain the amplification products of 9 HLA gene subtypes; The hybridization signals of the needles can be used to determine the subtypes of each HLA gene.

[0023] The primers and probe sequences involved in the present invention are as follows in Table 1:

[0024] Table 1. PCR primer sequences for multiplex amplification of 9 HLA subtypes

[0025]

[0026] The 5' ends of the downstream primers are all modified with Biotin. Three bases of GCG were adde...

Embodiment 2

[0042] Comparison test under different conditions of embodiment 2

[0043] In the multiplex PCR amplification system of the present invention, the upstream primer is set as a rate-limiting primer, additional bases are added to improve the Tm value, and the upstream primer is 4-8°C higher than the Tm value of the downstream primer; and the concentration of the rate-limiting primer is only 1 / 40 of the downstream primers, so in PCR amplification, more single strands prepared by downstream primers can be obtained. The traditional method of preparing single strands by PCR generally adjusts the ratio of upstream and downstream primers. Only after the primers with a small proportion are used up can a certain amount of single strands be obtained by single-primer amplification on the excess primers. The present invention combines by increasing the annealing temperature of rate-limiting primers and adjusting the ratio of upstream and downstream primers, and can prepare more single stran...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to an HLA subtype detection kit. The kit comprises primers and probes of multiplex PCR amplification for detecting a plurality of HLA subtypes, the amount ratio of each pair of upstream and downstream primers for HLA subtypes is 1:20 to 1:50, and the Tm value of the upstream primer of each pair is 4-8 DEG C higher than the Tm value of the downstream primer. According to the present invention, by the combination of increasing the annealing temperature of a rate-limiting primer and adjusting the ratio of the upstream and downstream primers, more single strands can be prepared without losing the PCR amplification efficiency, compared with traditional single-strand preparation methods, so that the hybridization signals of the probes can be significantly improved comparedwith the traditional multiple single-strand preparation PCR, so as to improve the sensitivity and specificity of the detection.

Description

technical field [0001] The invention relates to the field of molecular diagnosis, in particular to an HLA subtype detection kit and its application. Background technique [0002] Epilepsy is the second most common disease in neurology. There are over ten million epilepsy patients in my country, and about 400,000 new cases are added each year. Drug therapy is the main treatment for epilepsy. Most patients need to take medicine for a long time, but about 16% of patients have skin allergic reactions. Including mild maculopapular exanthema (MPE) and severe Stevens-Johnson syndrome (Stevens-Johnson syndrome, SJS) and toxic epidermal necrolysis (toxic epidermal necrolysis, TEN), significantly affecting the quality of life Exfoliative dermatitis occurs when it is severe, usually with ulcers on the mucous membranes (eyes, mouth, genitals) of the whole body, blisters on the skin, and even detachment of the whole body's epidermis, just like a large area of ​​burns, the fatality is as hi...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12Q1/6881C12Q1/6858
CPCC12Q1/6883C12Q1/6881C12Q1/6858C12Q2600/106C12Q2600/16C12Q2600/156C12Q2531/113C12Q2537/143
Inventor 廖卫平石弈武
Owner THE SECOND AFFILIATED HOSPITAL OF GUANGZHOU MEDICAL UNIV