肺病患者呼吸道曲霉随机扩增多态性DNA基因型分析

目的：了解肺病患者呼吸道曲霉基因分型状况,寻找肺部曲霉感染的可能来源,为曲霉感染的防治提供参考。方法：选择有呼吸道症状的患者925例作受试对象,收集其鼻腔、咽部分泌物及支气管肺泡灌洗液（bronchoal veolar lavage fluid,BALF）进行真菌培养、鉴定及随机扩增多态性DNA（random amplified polymorphic DNA,RAPD）基因分型分析。结果：在烟曲霉、黄曲霉和黑曲霉的RAPD带形图中,扩增条带数在1～11条之间,片断大小在150bp～2kb不等,不同菌株之间扩增条带的数量和扩增片断的大小均存在差异,即使带型完全一致的菌株,有些片断也存在亮度的差异。对分离自同一患者呼吸道不同部位的同种曲霉种内相似性比较的结果显示：虽然来自同一患者呼吸道不同部位的同种曲霉的形态学特征相同,但是基因型特征却存在差异。结论：通过呼吸作用进入肺组织内部的曲霉与患者鼻腔定植的同种曲霉存在差异,提示肺部曲霉的感染可能来源于患者周围的环境中,有效的控制环境中曲霉的污染程度是预防肺部曲霉感染的最有利的手段。

Analysis of Respiratory Tract Aspergilli s Genotype by Using the RAPD Technic in Patients with Pulmonary Diseases

Objective： To supervise the genotype of respiratory tract aspergilli in patients with pulmonary diseases, and search the probably infection source of pulmonary aspergilli. Methods： Through detecting 925 cases of pulmonary diseases＇ patients, the secretion of nasopharynx and BALF had been collected to be cultured, identified and analyzed the genotype of the same kind ofaspergilli which were isolated from the patients＇ different parts of the respiratory tract by using the RAPD technic. Results： It has been found that in the RAPD electrophoresis maps of A. fumigatus , A. flavus and A. niger, the number of amplification bands ranged from 1 to 11, in which the smallest fragment is about 150bp and the largest more than 2kb. Among the different strains, the quantity and the size of the amplification bands were all in differences. Even though the strains exactly had the same bands, the difference of brightness also existed in some fragments. Subsequently, the similarity of intraspecies that isolated from the aspergilli implanting in the different parts of the patients＇ respiratory tract was compared. The results indicated that although the same kind of aspergilli existing in the different parts of the patients＇ respiratory tract had the same morphological features, they were differences in characteristics of the genotype. Conclusion： It suggested that the aspergilli implanting in the internal pulmonary tissue through respiration differed from the same kind of aspergilli existing in the nasal cavity of the patients. The above demonstrated that the infection source of pulmonary aspergilli should probably originate from the patients＇ surrounding environments. So effectively controlling the aspergilli pollution of the surroundings is the best means to prevent pulmonary aspergillosis.