mLES法培养人芽囊原虫的实验观察  

Experimental study on culturing Blastocystis hominis using the mLES method

作　　者：孙煦苧 廖德君 刘静[1] 傅晓茵[1] 裴培 廖振捷 刘登宇[1] 辛致炜 SUN Xu-ning;LIAO De-jun;LIU Jing;FU Xiao-yin;PEI Pei;LIAO Zhen-jie;LIU Deng-yu;XIN Zhi-wei(Department of Parasitology, School of Preclinical Medicine, Guangxi Medical University, Nanning 530021, China;Department of Parasitology, Qiannan Medical College for Nationalities;Department of Parasitology, Institute of Microbiology and Immunology, College of Medicine, National Cheng Kung University)

机构地区：[1]广西医科大学病原生物学系,广西南宁530021 [2]黔南民族医学高等专科学校病原生物学教研室 [3]台湾成功大学医学院寄生虫学科

出　　处：《中国病原生物学杂志》2018年第3期259-262,266,共5页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.81360256);广西高校科研项目(No.YB2014063);广西自然科学基金项目(No.2017GXNSFAA198124)

摘　　要：目的建立一种简便、灵敏的可用于人芽囊原虫(Blastocystis hominis,Bh)感染诊断实验室方法。方法比较改良洛氏-鸡蛋-血清双相培养法(modified Locke-egg serum medium culture method,mLES培养法)与碘液直接涂片法对Bh的检出率;比较普通洛氏-鸡蛋-血清双相培养基(Locke-egg serum medium culture method,LES培养法)与mLES培养法中Bh形态、最低检出限、生长曲线及厌氧条件形成时间。结果检查397份临床粪便标本,mLES培养法Bh阳性率为5.54%(22/397),碘染法阳性率为1.01%(4/397),差异有统计学意义(P<0.01)。中央空泡型是Bh存在两种培养基中的主要形态。mLES培养法48hBh最低检出限(10~2/5ml)优于LES培养法(10~3/5ml),重复测量方差分析表明组内Bh密度在不同时间点有差异、时间与分组交互作用及培养基种类的差异有统计学意义(P<0.05);除了第6d,其余各时间点两种培养基中人芽囊原虫生长密度差异均有统计学意义(P<0.05),mLES培养法前3dBh密度高于LES培养法。mLES培养法先于普通培养基达到厌氧环境。结论 mLES培养法简便、灵敏,可用于诊断Bh的体外培养检查。Objectives To design a simple and sensitive method of in vitro culturing with which to diagnose Blastocystis hominis. Method Modified Locke-egg serum medium culturing（mLES culturing）and an iodine direct smear were compared in terms of the rate at which they detected B.hominis.In addition,the minimum detection limit 48 hafter infection,the growth curve,and the time needed to create anaerobic conditions for the 2 techniques were compared. Results mLES culturing（22/397）detected B.hominis at a higher rate than the iodine direct smear（4/397）,and the rate of detection differed significantly（P〈0.01）.In the 2 media,B.hominis was mainly present in vacuolar form（a spherical cell with a central vacuole）.The minimum detection limit of mLES culturing（102/5 ml）after 48 hwas higher than that in LES culturing（103/5 ml）.Repeated measures ANOVA indicated that there was significant difference in B.hominis density at different time points in each group and a significant difference in the interaction of time＊groups and grouping factors（type of media）（P〈0.05）.Moreover,results indicated that the density of B.hominis was significantly higher in the two media at all time points（P〈0.05）except day 6（P=0.327〉0.05）.The density of growth differed significantly（P〈0.05）,and the density of B.hominis in mLES culturing was higher than that in LES culturing in the first 3 days.mLES culturing produced an anaerobic environment faster than LES culturing. Conclusion mLES culturing is a simpler,more convenient,and more sensitive form of culturing that can be used to diagnose B.hominis.

关 键 词：人芽囊原虫 培养法 诊断 

分 类 号：R382[医药卫生—医学寄生虫学]