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Human leukocyte antigen gene detection kit for screening skin adverse reactions caused by clindamycin

An adverse reaction and kit technology, which is applied in biological testing, microbial determination/inspection, measurement devices, etc., and can solve problems such as no kit reports.

Active Publication Date: 2020-11-20
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the relevant kit

Method used

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  • Human leukocyte antigen gene detection kit for screening skin adverse reactions caused by clindamycin
  • Human leukocyte antigen gene detection kit for screening skin adverse reactions caused by clindamycin
  • Human leukocyte antigen gene detection kit for screening skin adverse reactions caused by clindamycin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1: Collection and extraction of genes

[0093] Patients with drug eruption caused by clindamycin came from Huashan Hospital Affiliated to Fudan University in Shanghai, China. They participated in the research of this subject under the premise of informed consent, collected blood, and signed an informed consent form. All enrolled patients were excluded from other diagnoses such as viral eruption and toxic erythema through medical history and skin pathological examination. All patients took clindamycin intravenously or orally within 2 months before the onset of rash. There are two groups of controls in this experiment. One is 283 healthy people from the human MHC database (dbMHC). In the lindamycin-resistant group, all patients received clindamycin intravenously for more than 2 months without any drug reaction;

[0094] Take 300 ul of sample blood, extract DNA according to the kit instructions, and detect the DNA concentration and purity with an ultraviolet spec...

Embodiment 2

[0095] Example 2: Detection of HLA genotyping

[0096] The present invention adopts PCR-SSO method (recommended Kit-One Lambda, CA, USA) for HLA genotyping. Its principle is to amplify the polymorphic region of HLA first, carry out isotope or non-isotope labeling on the PCR products during the amplification process, and then design a series of oligonucleotide probes for the PCR amplification products according to the principle of base pairing and immobilize them in Finally, the PCR product was hybridized with the probe on the membrane, and autoradiography was used to judge the result according to the signal. HLA genotyping was performed according to the standard steps of the kit. (i.e. DNA samples, substrates, Taq enzymes, and primers are mixed and evenly mixed, added to the amplification plate, amplified according to the conditions in the kit instructions, and the amplified products are hybridized, stained, and plate read). Results were analyzed by HLA Fusion software (On...

Embodiment 3

[0097] Example 3: HLA-B*51:01 is related to drug eruption caused by clindamycin

[0098] Statistics: SPSS20.0 was used to calculate Odds Ratio (OR) and its 95% confidence interval, P value was corrected by Bonferroni, Fisher's exact test was used for statistical analysis, and the statistical significance level was set at P less than 0.05.

[0099] The results showed that: 12 patients with clindamycin-induced drug eruption were collected, and 26 cases were in the clindamycin-resistant group. There were statistically significant differences between the clindamycin-resistant group (as shown in table 1) and the clindamycin-resistant group (as shown in table 2); the binding model of HLA-B*51:01 and clindamycin figure 1 Shown (model diagram of clindamycin binding to sHLA-B*51:01).

[0100] Table 1. Positive ratio of HLA-B*51:01 in the drug eruption group caused by clindamycin and the normal control group

[0101]

[0102] *The difference is statistically significant (P<0.05). ...

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Abstract

The invention belongs to the technical field of biological medicine, and relates to a human leukocyte antigen HLA-B*51: 01 gene detection kit. An HLA-B*51: 01 gene can be used as a marker gene for predicting clindamycin-induced drug rash. The invention provides the application of the detection kit to typing of the HLA-B*51: 01 gene in a sample. The application comprises the following steps: extracting DNA from obtained peripheral blood of a patient, and detecting the HLA-B*51: 01 gene by a current method, such as a sequence-specific oligonucleotide probe (PCR-SSO) method. The HLA-B*51: 01 gene detection kit provided by the invention can be applied to screening and sifting of targeted drugs for treating the clindamycin-induced drug rash before the use of clindamycin; screening application can guide clinical drug use so as to reduce incidence of the clindamycin-induced drug rash; selecting application mainly acts on an HLA-B*51: 01 molecule in a targeted manner so as to block interaction between the HLA-B*51: 01 molecule and clindamycin or other metabolites during the drug rash.

Description

technical field [0001] The invention belongs to the field of biomedicine and reagent detection, and relates to a human leukocyte antigen gene detection kit. It specifically involves the human leukocyte antigen gene - HLA-B*51:01 gene, which is related to the incidence of skin adverse drug reactions such as eruptive drug eruption, urticaria, fixed erythema drug eruption, and erythema multiforme caused by clindamycin . Background technique [0002] Clindamycin is one of the twenty most important antibiotics (J. et al., Lincomycin, clindamycin and their applications. Appl Microbiol Biotechnol, 2004.64: p.455–64.), is a commonly used drug in clinical practice, and is effective in Gram-positive bacteria, anaerobic Gram-negative bacteria, some native It plays an important role in the treatment of animals and fungal infections (Guay D. Update on clindamycin in the management of bacterial, fungal and protozoal infections. Expert Opin Pharmacother, 2007.8(14): p.2401-44. Frankel R...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883G01N33/68
CPCC12Q1/6883C12Q2600/106C12Q2600/156G01N33/6893G01N2333/705
Inventor 骆肖群邢清和杨颖陈圣安杨凡萍
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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