疟原虫体外培养过程中污染微生物鉴定及污染环节探讨

Identifications of the contaminated microbes in culture of Plasmodium falciparum in vitro and discussion on the resources and steps of contamination

目的鉴定体外培养疟原虫过程中污染的微生物种类，探讨污染微生物来源以及疟原虫培养过程中的污染环节。方法采用CO2混合气体法人工培养恶性疟原虫，并对污染的培养物进行细菌鉴定；分别采用表面采样法和自然沉降法对操作者手指部和实验室内空气进行微生物采样、培养、鉴定。结果培养过程中污染的微生物为嗜麦芽寡养单胞菌（Stenotrophomonas maltophilia）、鲍曼氏／醋酸钙不动杆菌复合体（Acinetobacter baumannii／calcoaceticus complex）和鲁氏不动杆菌[Acinetobacter lwoffii（acilwo）]，均为革兰阴性杆菌（G。），且属于多重耐药菌株；实验室空气中微生物数量在人员操作前后差异极大，操作前共检测到6个微生物菌落，操作后共检测到101个，紫外线照射后检测到4个。结论体外培养恶性疟原虫时污染微生物可能来源于操作者身体所携带，出现污染的环节最可能是在复苏过程。

Objective To identify species of the microbes contaminated in culture of Plasmodium falciparum in vitro,and to preliminarily discuss the sources of the contaminated microbes, as well as the most likely contamination steps. Methods 5% carbon dioxide mixture gas method was used for cultivation of P. falciparum in vitro. The microorganisms were collected, cultured and identified if the culture medium was contaminated. The microorganisms in the air samples of laboratory were collected by the natural sedimentation method, then cultured and identified, too. The microorganisms on surface of the operator＇s right hand were also collected before and after washing, then cultured and identified. Results The microbes from contaminated culture medium were Stenotrophomonas maltophilia, Acinetobacter baumannii/calcoaceticus complex and Acinetobacter lwoffii （acilwo）. They are Gram＇s negative bacillus （G-b）, and belong to the multiple drug resistance strains. The numbers of microbes from air samples in laboratory were significantly different before and after operation. The microbe colonies collected in the air samples before and after operation and after ultraviolet radiation were 6, 101 and 4, respectively. Conclusion The source of microbes＇ likely stems from the body of the operator, the contamination step is most probable in the setp of reviving in cultivation of P. falciparum.