Objective To evaluate the significance of several Dermatophagoides pteronyssinus allergen extracts for skin prick test(SPT) in patients allergic to Dermatophagoides pteronyssinus.Methods Two hundred and nineteen patients enrolled in Peking Union Medical College Hospital underwent SPT and serum specific IgE assay to detect the Dermatophagoides pteronyssinus allergen.Three kinds of house dust mite allergen extracts were used for SPT,including the Dermatophagoides pteronyssinus extract prepared by our laboratory(group A),standardized Dermatophagoides pteronyssinus extract(group B),and mixed extracts of Dermatophagoides pteronyssinus and Dermatophagoides farinae(group C).Human serum specific IgE result was regarded as the reference standard for diagnosis of Dermatophagoides pteronyssinus allergy.The receiver operating characteristic(ROC) curve was used to evaluate the diagnostic performance of SPT with the extracts of three groups.Results SPT results showed that the median wheal diameter of group A,group B,and group C was 0.43,0.35,and 0.28 cm,respectively,with significant difference among three groups(P<0.05).The difference was significant between group A and B(P<0.01) as well as group A and C(P<0.01),but not between group B and C(P>0.05).There was no local urticaria or systemic allergic reactions following the procedure of SPT.Local reaction was observed in 5 patients and delayed reaction was in 2 patients of group A.As for group B and C,local reaction occurred in 3 cases and delayed reaction in 2 cases in each group.The area under ROC curve of SPT with extract in group A,group B,and group C was 0.765,0.801,and 0.782,respectively.Based on the detection results of serum specific IgE,the sensitivity of SPT in diagnosis of Dermato-phagoides pteronyssinus allergy with extract of group A,group B,and group C was 92.4%,87.0%,and 81.5%,and the specificity was 60.6%,73.2%,and 74.8%,respectively.Conclusion The Dermatophagoides pteronyssinus extract for SPT prepared by our laboratory offers good sensitivity and specificity co
Objective To explore an effective method of Dermatophagoides pteronyssinus protein extraction suitable for two-dimensional electrophoresis (2-DE) analysis. Methods The extracts of Dermatophagoides pteronyssinus were prepared with Coca's solution, lysis buffer of 2-DE, and Trizol reagent, respectively. Bicinchoninic acid (BCA) assay was used to determine the total protein concentration of the samples. The efficiency of different protein extraction methods were evaluated with 2-DE analysis. Results The concentrations of extracted protein by methods of Coca's solution, lysis buffer, and Trizol reagent were 0.63 g/L,0.90 g/L,and 0.80 g/L, respectively. The 2-DE analysis results showed that some protein spots in low molecular weight (LMW) range could be detected with the Coca's solution method. With the lysis buffer of 2-DE method, more protein spots in LMW range could be detected, while the medium molecular weight (MMW) protein spots were absent. Several MMW protein spots (174-178 kD and 133 kD) and more LMW protein spots were detected with Trizol reagent method. Conclusions Among Coca's solution, lysis buffer of 2-DE, and Trizol reagent, the concentration of extracted protein of Dermatophagoides pteronyssinus by lysis buffer of 2-DE is the highest. However, most protein components of Dermatophagoides pteronyssinus purified mite bodies can be extracted by Trizol reagent, which may generally reflect the whole profile of Dermatophagoides pteronyssinus allergens.
Jin-lu SunHong-yu ZhangZhi-yi GuoWan-tao YingXiao-hong QianJing-lan Wang
