| Objective: Chronic urticaria is the most common chronic allergic disease with a high incidence and a long duration.This disease greatly interferes with the patient's life and work for the severe itching.Finding allergens is the key to diagnosis and treatment.Dermatophagoides pteronyssinus is an important allergen in chronic urticaria.The detection of serum specific Ig E(s Ig E),as a safe,effective and convenient detection method,is widely used in the diagnosis of allergic diseases.However,the inconsistence of the clinical and laboratory diagnosis is founded sometimes.In order to optimize the s Ig E detection system of dermatophagoides pteronyssinus in patients with chronic urticaria,we designed the following schemes to purifying the proteins from dermatophagoides pteronyssinus.Methods: 1 An epidemiological investigation was carried out on patients with allergic diseases in the dermatology of dermatology,the Fourth Hospital of Hebei Medical University from January 2018 to January 2019,who were carried out the detection of s Ig E.2 The component of Dermatophagoides pteronyssinus was analyzed by Sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE).3 The serum pool of chronic urticaria patients,who had the inconsistent result of serum s Ig E test and clinical manifestations for Dermatophagoides pteronyssinus was selected for Western-blot(WB)and proteomic analysis.4 Further analysis of the Dermatophagoides pteronyssinus were performanced by high performance liquid chromatography(HPLC),followed by SDS-PAGE and WB of each peak solution.5 The results of the main peak 2 by slot-blot(SB)and enzyme-linked immunosorbent assay(ELISA)was compared to that of serum s Ig E detection system,respectively.Results: 1 In our hospital,the main allergens of patients visiting in our hospital are dust mite,egg white and milk.2 Epidemiological investigations: 2.1 In our hospital,from January 2018 to January 2019,totally there were 2665 patients registered,including 1032 males and 1633 females,and the average age is 33.4±10.5.The main diseases are urticaria,dermatitis and eczema,including chronic urticaria 644 cases,ranking first.There are 235 patients with chronic urticaria were suspected to dust mite,accounting for the highest proportion.2.2 Comparing the results of epidemiological investigation and serum allergen s Ig E detection in the 235 patients with chronic urticaria,we found that 87 cases of patients diagnosed as chronic urticaria suspected of dust mite allergy had serum allergen s Ig E detection results inconsistent with clinical manifestations.3 By SDS-PAGE,16 bands were found,the molecular weight are respectively estimated to be 129 k Da,117 k Da,105 k Da,59.1k Da,56.8k Da,49.0k Da,43.7k Da,39.1k Da,35.5k Da,28.6k Da,25.7k Da,22.4k Da,19.6k Da,16.7k Da,15.5k Da and 13.9k Da.4 By WB,many bands were found.The bands of 59.1k Da,49.0k Da-39.1k Da,35.5k Da,28.6k Da,25.7k Da,22.4k Da and 15.5k Da were obvious,considered as Der p4,Der p9,Der p10,Der p7,Der p1,Der p3 and Der p2.5 Seven main peaks were detected by HPLC chromatogram for the extract of Dermatophagoides pteronyssinus.By analyzing the results of SDS-PAGE and WB,it can be seen that the more obvious reaction bands of the main peak 2 were 61.6k Da,45.5k Da,35.8 k Da,32.6k Da,26.5k Da,24.2k Da and 16.3k Da,considering as Der p4,Der p9,Der p10,Der p7,Der p1,Der p3 and Der p2.6 The results of the main peak 2 by slot-blot(SB)or enzyme-linked immunosorbent assay(ELISA)were more accurate compared to that of serum s Ig E detection system.Conclusions: 1.In our hospital the main allergens are dust mite,egg white and milk.2.In the chronic urticaria patients,the major allergens of Dermatophagoides pteronyssinus were considered to be Der p4,Der p9,Der p10,Der p7,Der p1,Der p3,Der p3 and Der p2.3.Using Dermatophagoides pteronyssinus protein purified by HPLC,the two methods of SB and ELISA all can be optimize the detection results of Dermatophagoides pteronyssinus allergen. |
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