The acaricidal activities of fourteen essential oils and fourteen of their major monoterpenoids were tested against house dust mites Dermatophagoides pteronyssinus. Five concentrations were used over two different tim...The acaricidal activities of fourteen essential oils and fourteen of their major monoterpenoids were tested against house dust mites Dermatophagoides pteronyssinus. Five concentrations were used over two different time intervals 24 and 48 h under laboratory conditions. In general, it was noticed that the acaricidal effect based on LC50 of either essential oils or monoterpenoids against the mite was time dependant. The LCso values were decreased by increasing of exposure time. Clove, matrecary, chenopodium, rosemary, eucalyptus and caraway oils were shown to have high activity. As for the monoterpenoids, cinnamaldehyde and chlorothymol were found to be the most effective followed by citronellol. This study suggests the use of the essential oils and their major constituents as ecofriendly biodegradable agents for the control of house dust mite, D. pteronyssinus.展开更多
Obiective To explore an effective method of Dermatophagoides pteronyssinus protein extraction suitable for two-dimensional electrophoresis （2-DE） analysis. Methods The extracts of Dermatophagoides pteronyssinus wer...Obiective 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 withTrizol 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.展开更多
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 patie...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 comparable to commercially available allergen extracts,and it may be an appropriate candidate for clinical screening and diagnosis of Dermatophagoides pteronyssinus allergy.展开更多
Objective:To examined the immediate and 24 hours post-irradiation germicidal effects of UV-C lamp on.eggs and adults of house dust mites Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermatophagoides farinae(D.fa...Objective:To examined the immediate and 24 hours post-irradiation germicidal effects of UV-C lamp on.eggs and adults of house dust mites Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermatophagoides farinae(D.farinae).Methods:This study investigated the immediate and24 hours post irradiation mortalities of adult mites exposed to UV-C at different exposure times(5mins,10 mins,15 mins,20 mins,30 mins and 60 mins)and distances(10 cm,25 cm,33 cm,45 cm and 55 cm).Fresh eggs of the 2 dust mites were also irradiated at 10,35 and 55 cm for 0.5,1,2,3,and 5 minutes,and observed daily post-irradiation for up to 7 days.Results:Highest immediate mortality of 100%occurred with direct irradiation al 10 cm distance from UV-C lamp and for 60mins,for both species of mites.The post 24 hours mean mortality rates were(58.4±17.4)%for D.pteronyssinus and(27.7±9.7)%for D.fitrinae when irradiated for 1 hour at 55 cm distance under UV-C lamp.When mites were irradiated in the presence of culture media,the highest mortality rates were lower compared to the direct irradiation;at 10 cm distance and 60 mins exposure,the mean mortality was(74.0±6.8)%for D.pteronyssinus and(70.3±6.7)%for D.farinae.Egg hatchability for both species of mites was also notably reduced by greater than 50%following irradiation.Conclusions:Ultraviolet C irradiation is lethal to an array of organisms by damaging their nucleic acids(DNA and RNA).This study demonstrates the increasing mite mortalities with increasing exposure times and decreasing distances.展开更多
Objective:To investigate the short and long tenn efficacy of a commercial air ionizer in killing Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermalophagoides farinae(D.farinae)mites.Methods:The effect of a comm...Objective:To investigate the short and long tenn efficacy of a commercial air ionizer in killing Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermalophagoides farinae(D.farinae)mites.Methods:The effect of a commercial ionizer on D.pteronyssinus and D.farinae was evaluated in the laboratory,using a specially designed test.Mortality was assessed after 6,16and 24 hours for direct exposure and after 24,36,48,60 and 72 hours for exposure in simulated mattress.New batches of mites were used for each exposure time.Results:LT_(50)for direct exposure of ionizer was 10 hours for D.pteronyssinus and 18 hours for D.farinae.The LT_(50)for exposure in simulated mattress was 132 hours or 5.5 days for D.pteronyssinus and 72 hours or 3days for D.farinae.LT_(95)for direct exposure of ionizer was 36 hours for D.pteronyssinus and D.farinae.Meanwhile,the LT_(95)for exposure in simulated mattress was 956 hours or 39.8 days for D.pteronyssinus and 403 hours or 16.8 days for D.farinae.Conclusions:This study demonstrates the increasing mite mortalities with increasing exposure time of a commercial ionizer and suggests that negative ions produced by an ionizer kill dust mites and can be used to reduce natural mile populations on exposed surfaces such as floors,clothes,curtains,etc.However,there is reduced efficacy on mites inside stuffed materials as in mattresses and furniture.展开更多
【目的】利用扫描电子显微镜(scanning electron microscopy,SEM)观察屋尘螨Dermatophagoides pteronyssinus颚体、躯体、外生殖器及足等的形态结构。【方法】从床尘、枕尘中采集屋尘螨,分离出雌雄成螨,在体视显微镜下清洗处理活螨后,用...【目的】利用扫描电子显微镜(scanning electron microscopy,SEM)观察屋尘螨Dermatophagoides pteronyssinus颚体、躯体、外生殖器及足等的形态结构。【方法】从床尘、枕尘中采集屋尘螨,分离出雌雄成螨,在体视显微镜下清洗处理活螨后,用SEM观察其外部形态特征。【结果】SEM照片显示,屋尘螨螯肢钳状,须肢扁平;体表具细密皮纹,似指纹状,纹间距小于2μm。外生殖器位于腹面正中,雌螨为产卵孔,雄螨生殖孔具1对叶状生殖盖。肛门呈纵行裂孔,雄螨具2个肛吸盘。雌螨足跗节末端各具爪垫1个,雄螨跗节Ⅳ具2个吸盘。【结论】本研究观察结果为尘螨鉴定提供了更多依据。展开更多
目的评价标准化屋尘螨过敏原淋巴免疫治疗过敏性哮喘的临床疗效及安全性。方法 72例屋尘螨致敏性哮喘患者,随机分为治疗组和对照组,每组各36例,对照组给予布地奈德气雾剂和沙丁胺醇气雾剂治疗;治疗组在上述药物治疗同时,行超声引导下分...目的评价标准化屋尘螨过敏原淋巴免疫治疗过敏性哮喘的临床疗效及安全性。方法 72例屋尘螨致敏性哮喘患者,随机分为治疗组和对照组,每组各36例,对照组给予布地奈德气雾剂和沙丁胺醇气雾剂治疗;治疗组在上述药物治疗同时,行超声引导下分别于0、4、8、12、16和20周行腹股沟淋巴结内注射屋尘螨过敏原制剂,除0周用药剂量为100 SQ-U外,其余5次应用剂量均为1000 SQ-U。治疗前后两组患者分别进行血清屋尘螨特异性IgE(sIgE)检测、哮喘控制测试(asthma control test,ACT),药物用量评分、肺功能[第1秒用力呼气容积占预计值的百分比(percentage of forced expiratory volume in first second to predicted value,FEV1%)、第1秒用力呼气容积与用力肺活量比值(ratio of forced expiratory volume in first second to forced vital capacity,FEV1/FVC)和峰流速占预计值的百分比(percentage of peak expiratory flow to predicted value,PEF%)]检测并记录患者疗程中的不良反应。结果治疗组32例患者完成了淋巴免疫治疗,2例因怀孕退出,2例失访;对照组32例按要求完成了对照观察,4例失访。治疗组患者治疗后4、8、12、16和20周ACT评分分别为(15.72±2.00)、(16.44±2.06)、(18.88±1.29)、(20.16±1.43)、(23.97±1.03),高于治疗前(0)周(15.34±2.15),差异有统计学意义(P<0.01);12、16和20周ACT评分均高于对照组同时间ACT评分[(16.63±2.12)、(16.81±2.10)、(16.87±2.30)],差异有统计学意义(P<0.01)。治疗后肺功能检测(FEV1%、FEV1/FVC和PEF%)分别为(74.81±5.91)、(77.31±5.43)、(74.09±4.71),均明显高于治疗前各项[(57.06±7.49)、(60.31±7.04)、(50.81±6.57)]及治疗后对照组水平[(54.91±7.79)、(59.44±8.77)、(50.38±8.13)],差异有统计学意义(P<0.01)。治疗组治疗后药物用量评分(0.66±0.43)和血清屋尘螨sIgE浓度(1.69±0.92)kU/L明显低于治疗前[(2.19±0.78)、(2.73±1.09)kU/L],且明显低于对照组[(1.92±0.77)、(3.09±1.29)kU/L],差异均有统计学差异(P<0.01)。32例淋巴免疫治疗患者进行了192次浅表淋巴结注射,未发生局部不良反应和全身不良反应。结论过敏原淋巴免疫治疗不仅临床疗效显著,而且大大缩短了疗程,减少了变应原剂量、注射次数和不良反应的发生,为安全、有效的病因治疗方法。展开更多
目的：建立Der p 2（屋尘螨Ⅱ类变应原）杂交瘤细胞株，并对其分泌的Der p 2单克隆抗体进行鉴定．方法：用重组Der p 2蛋白作为免疫原，免疫BALB／c小鼠，取脾细胞与Sp2／0小鼠骨髓瘤细胞融合，经筛选和3次克隆化，制备出单克隆抗体．...目的：建立Der p 2（屋尘螨Ⅱ类变应原）杂交瘤细胞株，并对其分泌的Der p 2单克隆抗体进行鉴定．方法：用重组Der p 2蛋白作为免疫原，免疫BALB／c小鼠，取脾细胞与Sp2／0小鼠骨髓瘤细胞融合，经筛选和3次克隆化，制备出单克隆抗体．采用间接ELISA法、Western blot法确定单克隆抗体的免疫球蛋白的类型和亚类、相对亲和力、特异性，对单克隆抗体进行鉴定．结果：获得3株能稳定分泌高效价Der p 2单克隆抗体的杂交瘤细胞株，186为IgG1类，1G11、4B1为IgG2a类．间接ELISA法检测细胞上清效价为10^4～10^5，腹水效价为10^5～10^6．Western blot试验证明3株单抗与Der p 2蛋白均有特异性反应，结论：成功制备了3株抗Der p 2的单克隆抗体，均具有良好的特异性和亲和力，为进一步建立Der p 2蛋白检测和纯化的方法奠定了基础．展开更多
文摘The acaricidal activities of fourteen essential oils and fourteen of their major monoterpenoids were tested against house dust mites Dermatophagoides pteronyssinus. Five concentrations were used over two different time intervals 24 and 48 h under laboratory conditions. In general, it was noticed that the acaricidal effect based on LC50 of either essential oils or monoterpenoids against the mite was time dependant. The LCso values were decreased by increasing of exposure time. Clove, matrecary, chenopodium, rosemary, eucalyptus and caraway oils were shown to have high activity. As for the monoterpenoids, cinnamaldehyde and chlorothymol were found to be the most effective followed by citronellol. This study suggests the use of the essential oils and their major constituents as ecofriendly biodegradable agents for the control of house dust mite, D. pteronyssinus.
基金Supported by grants from the Ministry of Science and Technology of China (2008BAI59B04, 2003AA2Z3502)National Natural Science Foundation of China (30671943)
文摘Obiective 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 withTrizol 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.
基金Supported by the National Natural Science Foundation of China (30671943)the National Scientific Supporting Foundation of China (2008BAI59B04)the National High Technology Research and Development Program of China (863 Program) (2003AA2Z3502)
文摘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 comparable to commercially available allergen extracts,and it may be an appropriate candidate for clinical screening and diagnosis of Dermatophagoides pteronyssinus allergy.
文摘Objective:To examined the immediate and 24 hours post-irradiation germicidal effects of UV-C lamp on.eggs and adults of house dust mites Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermatophagoides farinae(D.farinae).Methods:This study investigated the immediate and24 hours post irradiation mortalities of adult mites exposed to UV-C at different exposure times(5mins,10 mins,15 mins,20 mins,30 mins and 60 mins)and distances(10 cm,25 cm,33 cm,45 cm and 55 cm).Fresh eggs of the 2 dust mites were also irradiated at 10,35 and 55 cm for 0.5,1,2,3,and 5 minutes,and observed daily post-irradiation for up to 7 days.Results:Highest immediate mortality of 100%occurred with direct irradiation al 10 cm distance from UV-C lamp and for 60mins,for both species of mites.The post 24 hours mean mortality rates were(58.4±17.4)%for D.pteronyssinus and(27.7±9.7)%for D.fitrinae when irradiated for 1 hour at 55 cm distance under UV-C lamp.When mites were irradiated in the presence of culture media,the highest mortality rates were lower compared to the direct irradiation;at 10 cm distance and 60 mins exposure,the mean mortality was(74.0±6.8)%for D.pteronyssinus and(70.3±6.7)%for D.farinae.Egg hatchability for both species of mites was also notably reduced by greater than 50%following irradiation.Conclusions:Ultraviolet C irradiation is lethal to an array of organisms by damaging their nucleic acids(DNA and RNA).This study demonstrates the increasing mite mortalities with increasing exposure times and decreasing distances.
文摘Objective:To investigate the short and long tenn efficacy of a commercial air ionizer in killing Dermatophagoides pteronyssinus(D.pteronyssinus)and Dermalophagoides farinae(D.farinae)mites.Methods:The effect of a commercial ionizer on D.pteronyssinus and D.farinae was evaluated in the laboratory,using a specially designed test.Mortality was assessed after 6,16and 24 hours for direct exposure and after 24,36,48,60 and 72 hours for exposure in simulated mattress.New batches of mites were used for each exposure time.Results:LT_(50)for direct exposure of ionizer was 10 hours for D.pteronyssinus and 18 hours for D.farinae.The LT_(50)for exposure in simulated mattress was 132 hours or 5.5 days for D.pteronyssinus and 72 hours or 3days for D.farinae.LT_(95)for direct exposure of ionizer was 36 hours for D.pteronyssinus and D.farinae.Meanwhile,the LT_(95)for exposure in simulated mattress was 956 hours or 39.8 days for D.pteronyssinus and 403 hours or 16.8 days for D.farinae.Conclusions:This study demonstrates the increasing mite mortalities with increasing exposure time of a commercial ionizer and suggests that negative ions produced by an ionizer kill dust mites and can be used to reduce natural mile populations on exposed surfaces such as floors,clothes,curtains,etc.However,there is reduced efficacy on mites inside stuffed materials as in mattresses and furniture.
文摘【目的】利用扫描电子显微镜(scanning electron microscopy,SEM)观察屋尘螨Dermatophagoides pteronyssinus颚体、躯体、外生殖器及足等的形态结构。【方法】从床尘、枕尘中采集屋尘螨,分离出雌雄成螨,在体视显微镜下清洗处理活螨后,用SEM观察其外部形态特征。【结果】SEM照片显示,屋尘螨螯肢钳状,须肢扁平;体表具细密皮纹,似指纹状,纹间距小于2μm。外生殖器位于腹面正中,雌螨为产卵孔,雄螨生殖孔具1对叶状生殖盖。肛门呈纵行裂孔,雄螨具2个肛吸盘。雌螨足跗节末端各具爪垫1个,雄螨跗节Ⅳ具2个吸盘。【结论】本研究观察结果为尘螨鉴定提供了更多依据。
文摘目的评价标准化屋尘螨过敏原淋巴免疫治疗过敏性哮喘的临床疗效及安全性。方法 72例屋尘螨致敏性哮喘患者,随机分为治疗组和对照组,每组各36例,对照组给予布地奈德气雾剂和沙丁胺醇气雾剂治疗;治疗组在上述药物治疗同时,行超声引导下分别于0、4、8、12、16和20周行腹股沟淋巴结内注射屋尘螨过敏原制剂,除0周用药剂量为100 SQ-U外,其余5次应用剂量均为1000 SQ-U。治疗前后两组患者分别进行血清屋尘螨特异性IgE(sIgE)检测、哮喘控制测试(asthma control test,ACT),药物用量评分、肺功能[第1秒用力呼气容积占预计值的百分比(percentage of forced expiratory volume in first second to predicted value,FEV1%)、第1秒用力呼气容积与用力肺活量比值(ratio of forced expiratory volume in first second to forced vital capacity,FEV1/FVC)和峰流速占预计值的百分比(percentage of peak expiratory flow to predicted value,PEF%)]检测并记录患者疗程中的不良反应。结果治疗组32例患者完成了淋巴免疫治疗,2例因怀孕退出,2例失访;对照组32例按要求完成了对照观察,4例失访。治疗组患者治疗后4、8、12、16和20周ACT评分分别为(15.72±2.00)、(16.44±2.06)、(18.88±1.29)、(20.16±1.43)、(23.97±1.03),高于治疗前(0)周(15.34±2.15),差异有统计学意义(P<0.01);12、16和20周ACT评分均高于对照组同时间ACT评分[(16.63±2.12)、(16.81±2.10)、(16.87±2.30)],差异有统计学意义(P<0.01)。治疗后肺功能检测(FEV1%、FEV1/FVC和PEF%)分别为(74.81±5.91)、(77.31±5.43)、(74.09±4.71),均明显高于治疗前各项[(57.06±7.49)、(60.31±7.04)、(50.81±6.57)]及治疗后对照组水平[(54.91±7.79)、(59.44±8.77)、(50.38±8.13)],差异有统计学意义(P<0.01)。治疗组治疗后药物用量评分(0.66±0.43)和血清屋尘螨sIgE浓度(1.69±0.92)kU/L明显低于治疗前[(2.19±0.78)、(2.73±1.09)kU/L],且明显低于对照组[(1.92±0.77)、(3.09±1.29)kU/L],差异均有统计学差异(P<0.01)。32例淋巴免疫治疗患者进行了192次浅表淋巴结注射,未发生局部不良反应和全身不良反应。结论过敏原淋巴免疫治疗不仅临床疗效显著,而且大大缩短了疗程,减少了变应原剂量、注射次数和不良反应的发生,为安全、有效的病因治疗方法。
文摘目的：建立Der p 2（屋尘螨Ⅱ类变应原）杂交瘤细胞株，并对其分泌的Der p 2单克隆抗体进行鉴定．方法：用重组Der p 2蛋白作为免疫原，免疫BALB／c小鼠，取脾细胞与Sp2／0小鼠骨髓瘤细胞融合，经筛选和3次克隆化，制备出单克隆抗体．采用间接ELISA法、Western blot法确定单克隆抗体的免疫球蛋白的类型和亚类、相对亲和力、特异性，对单克隆抗体进行鉴定．结果：获得3株能稳定分泌高效价Der p 2单克隆抗体的杂交瘤细胞株，186为IgG1类，1G11、4B1为IgG2a类．间接ELISA法检测细胞上清效价为10^4～10^5，腹水效价为10^5～10^6．Western blot试验证明3株单抗与Der p 2蛋白均有特异性反应，结论：成功制备了3株抗Der p 2的单克隆抗体，均具有良好的特异性和亲和力，为进一步建立Der p 2蛋白检测和纯化的方法奠定了基础．
