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目的观察人芽囊原虫(Blastocystis hominis,Bh)在IMDM、RPMI1640以及传统的洛克氏液-鸡蛋-血清(LES)培养基中的生长情况,应用正交试验筛选人芽囊原虫的最适培养基及培养条件。方法将门诊病人Bh阳性粪便标本调匀后作虫体计数,分别接种于IMDM、RPMI1640及LES培养基进行培养,以正交试验分析培养基、血清浓度、pH值和接种虫体数4个因素对Bh体外培养效果的影响,以首次达虫体数量高峰时间、虫密度平均峰值、虫密度平均值的综合评分值K为指标,考核虫株的综合培养效果。结果对正交试验结果进行极差及方差分析,各因素对Bh培养效果的影响为:培养基>血清浓度>pH值>接种量。经SSR检验,IMDM、RPMI 1640及LES 3种培养基对Bh的培养效果差异有统计学意义(P<0.05),以IMDM培养基中培养综合评分K值最高,培养效果最优;RPMI 1640与LES培养基对Bh的培养效果差异无统计学意义(P>0.05)。培养基中加入血清0.5、1、1.5ml时培养效果差异有统计学意义(P<0.05),以血清量0.5ml培养效果最优;Bh在pH7.0、pH7.5、pH8.0条件下培养效果差异有统计学意义(P<0.05),在pH7.5和pH8之间培养效果差异无统计学意义(P>0.05)。结论含血清0.5ml、pH 7.5～8.0的IMDM单相培养基适宜Bh生长繁殖,可用于Bh体外长期培养。
Objective To observe the growth of Blastocystis hominis (Bh) in IMDM, RPMI1640 and traditional Lactobacillus casei-egg-serum (LES) medium. The optimal medium for screening Blastocystis hominis And culture conditions. Methods Bh-positive stool specimens from outpatients were evenly collected and counted for parasites. They were respectively inoculated into IMDM, RPMI1640 and LES medium for culture. The effects of medium, serum concentration, pH value and the number of inoculated parasites Bh in vitro culture effect, the first time the amount of peak number of larvae, the average peak density of insects, the average value of insect density integrated score K as an indicator to assess the integrated culture of insect strains. Results The range and variance of the orthogonal test results were analyzed. The influence of each factor on the culture effect of Bh was: medium> serum concentration> pH value> inoculum size. The results of SSR showed that there was significant difference in the culture effect of BMI between IMDM, RPMI 1640 and LES (P <0.05), and the highest comprehensive score (K) in IMDM culture medium with the best culture effect; RPMI 1640 and There was no significant difference in the culture effect of LES medium on Bh (P> 0.05). The culture effect of 0.5,1,1.5ml serum was statistically different (P <0.05), 0.5ml serum culture best; Bh pH7.0, pH7.5, pH8.0 conditions There was no significant difference in culture effect between pH7.5 and pH8 (P> 0.05). Conclusion IMDM single phase medium containing 0.5 ml of serum and pH 7.5-8.0 is suitable for Bh growth and reproduction and can be used for long-term culture of Bh in vitro.