Human leukocyte antigen gene detection kit and application thereof
A leukocyte antigen and gene detection technology, which is used in the screening of skin adverse drug reactions caused by metronidazole, a human leukocyte antigen gene detection kit, and a human leukocyte antigen gene-HLA-B*39:01 gene detection kit. Can solve problems such as unseen and unseen kit reports
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Embodiment 1
[0048] Example 1: Collection and extraction of genes
[0049] Patients with drug eruption caused by metronidazole were from Huashan Hospital Affiliated to Fudan University, Shanghai, China. Among them, the diagnosis of urticaria-type drug eruption needs to meet the clinical requirements of wheal-like rash, which cannot completely subside within 24 hours, and may or may not be accompanied by throat obstruction or paroxysmal abdominal pain; exanthematous drug eruption is a generalized symmetrically distributed red patch , and no mucosal involvement; fixed drug eruptions are single or multiple target lesions, and pigmentation remains after the rash improves (Roujeau JC. Clinical heterogeneity of drug hypersensitivity. Toxicology 209(2), 123–129(2005). S, Rzany B, Stern RS, Shear NH, Naldi L, Roujeau JC. Clinical classification of cases of toxic epidermal necrolysis, Stevens–Johnson syndrome, and erythema multiforme. Arch. Dermatol. 129(1), 92–96(1993)); On the premise of informe...
Embodiment 2
[0050] Example 2: Detection of HLA genotyping
[0051] The present invention adopts PCR-SSO method (recommended Kit-One Lambda, CA, USA) for HLA genotyping; the principle is to amplify the polymorphic region of HLA first, and perform isotope or non-isotope labeling on the PCR product during the amplification process, and then target the PCR amplification. According to the principle of base pairing, a series of oligonucleotide probes are designed and immobilized on the membrane, and finally the PCR product is hybridized with the probes on the membrane, and autoradiography is used to judge the result according to the signal; HLA genotyping is carried out according to the standard steps of the kit : Mix DNA samples, substrates, Taq enzymes, and primers and mix them evenly, add them to the amplification plate, amplify according to the conditions in the kit instructions, and perform hybridization, staining, and plate reading of the amplified products; through HLAFusion software (O...
Embodiment 3
[0052] Example 3: HLA-B*39:01 is related to drug eruption caused by metronidazole
[0053] Odds Ratio (OR) and its 95% confidence interval were calculated by using SPSS16.0, and Haldane’s correction was used when necessary, and chi-square test was used for statistical analysis, and the statistical significance level was set at P less than 0.05;
[0054] The results showed that 9 patients with drug eruption caused by metronidazole were collected, and the clinical features and HLA types of drug eruption caused by metronidazole are shown in Table 1;
[0055] Table 1
[0056]
[0057] The positive ratio of HLA-B*39:01 in patients with drug eruption caused by metronidazole compared with the positive carrier ratio of the normal control group is shown in Table 2; the difference between them is statistically significant.
[0058] Table 2
[0059]
[0060] *The difference is statistically significant (P<0.05).
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