【作者】 童华章；

【导师】 刘志刚；

【作者基本信息】 江西医学院 ， 病原生物学， 2003， 硕士

【摘要】 背景 互隔交链孢霉是一种常见且重要的气传致敏真菌变应原。目前国内尚未见有关互隔交链孢霉变应原分离纯化及免疫学特性的研究报道。目的 旨在对互隔交链孢霉主、次要变应原分离、鉴定和初步纯化，为真菌变应原的标准化提供理论基础。方法 本研究在前期对南昌地区气传致敏真菌调查的基础上，选择互隔交链孢霉作为研究对象，采用不同的提取方式对互隔交链孢霉进行提取；通过聚丙烯酰胺凝胶电泳（SDS-PAGE）对其蛋白组分进行分离；选用Western-blotting对其主要及次要变应原进行鉴定；通过DEAE Cellulose-52离子交换层析对互隔交链孢霉的主、次要变应原进行初步纯化。结果 本研究采用不同的提取方式得到互隔交链孢霉的粗浸液，通过SDS-PAGE主要分离到9种蛋白质组分；通过Western-blotting，发现40KD和 45KD的抗原为互隔交链孢霉主要变应原，13KD、57KD和71KD的抗原为次要变应原。通过DEAE Cellulose-52离子交换层析对互隔交链孢霉变应原进行初步纯化，得到主要及次要变应原的混合组分。结论 互隔交链孢霉的主要变应原成分代表了变应原主要致敏成分，对互隔交链孢霉变应原进行分离、鉴定和初步纯化，将为进一步研制标准化的真菌变应原制剂及探讨更安全、有效的诊断和治疗方法提供理论基础。更多还原

【Abstract】 Background Alternaria alternata is one of the most important and common fungal allergens in the world. Many articles about purification and identification of Alternaria allergens have been reported abroad so far. But few have been reported in our country.Objective Our research is to extract allergens of Alternaria alternata, to identify and partially purify the primary and the secondary allergens of Alternaria alternata, to provide theoretical basis for the standardization of fungal allergens.Methods On the basis of our previous investigation on aero-allergens in Nanchan city, Jiangxi province, we selected Alternaria alternata as our research subject. The Alternaria alternate allergens were extracted by different extraction methods , the protein allergen components were isolated by means of SDS-polyacrylamide gel electrophoresis (SDS-PAGE) . And its primary and secondary allergen proteins have been identified by Western-blotting. Then, primary and secondary allergen proteins of Alternaria alternata were purified by DEAE Cellulose-52 Ion Exchange Chromato-graphy (IEC).Their bioactivities were detected by Western-blotting.Results Several protein bands from Alternaria alternata extract have been isolated by means of SDS-polyacrylamide gel electro-phoresis (SDS-PAGE).The results show that the antigens with the<WP=5>molecular weight of 40KD and 45KD are the primary allergens of Alternaria alternata;Unique allergen protein bands of the Alternaria alternata were identified , whose molecular weights are 40KD and 45KD. And its primary and secondary allergen proteins have been identified by Western blotting.Then the primary and secondary allergen proteins of Alternaria alternata were purified by DEAE Cellulose-52 Ion Exchange Chromatography (IEC). Its bioactivity was detected by Western-blotting. Its molecular weight determined by SDS-PAGE was 40K Dalton and 45K Dalton.Conclusion The major allergens represent the main allergic components of Alternaria alternata. The isolation, identification and purification of the primary antigens of Alternaria alternata will provide theoretical basis for the development of standardized fungal allergen and explore safer and more effective diagnosis and treatment methods for the fungal allergic diseases.更多还原