周悦昌，　任丽娜，　王旭辉，　陈建文，　陈妙佩，　毛鸿忠，　韩立杰

不同培养条件下透析液和反渗水微生物监测的比较分析

周悦昌1，任丽娜1，王旭辉1，陈建文3，陈妙佩3，毛鸿忠3，韩立杰2

(1. 牡丹江巿第二人民医院检验科，黑龙江 牡丹江 157000；

2. 牡丹江巿第二人民医院血液净化中心，黑龙江 牡丹江 157000；

3. 屏基医疗财团法人屏东基督教医院教学研究部肾脏科研究小组，台湾 屏东 811)

摘要：目的比较不同培养基、孵育温度及培养时间的选择对透析液和反渗水微生物监测总菌落计数的差异，确认透析液和反渗水微生物监测的最佳培养条件。方法采用无菌透析液和无菌反渗水配置模拟污染水样本，倾注法接种于伊红美蓝琼脂(EMB)、大豆酪蛋白琼脂(TSA)、胰蛋白胨葡萄糖浸液(TGEA)培养基，于25℃和35℃分别进行培养，并记录培养48、72 h及7 d的生长情况。结果不论是25℃还是35℃，TGEA的生长情况均最佳且最接近预期菌落数(P<0.05)；各种培养基培养72 h比48 h生长好(P<0.05)；培养7 d后的菌落数与培养72 h比较差异无统计学意义(P>0.05)。 结论使用倾注平板法及TGEA培养基对透析液和反渗水进行微生物监测，需同时在25℃及35℃条件下培养至少72 h，方可产生有效的监测报告，确保患者的透析安全。

关键词：血液透析液；反渗水；总菌落数计算

Comparison analysis of microbiological monitoring between hemodialysate and reverse osmosis water under different mediaZHOUYuechang1，RENLina1，WANGXuhui1，CHENJianwen3，CHENMiaopei3，MAOHongzhong3，HANLijie2

.(1.DepartmentofClinicalLaboratory，theSecondPeople′sHospitalofMudanjang，HeilongjiangMudanjiang157000,China； 2.BloodPurificationCenter，theSecondPeople′sHospitalofMudanjang，HeilongjiangMudanjiang157000,China； 3.TeamofNephrologicalResearch,DepartmentofMedicalResearch，PingtungChristianHospital，TaiwanPingtung811,China)

Abstract:ObjectiveTo compare the difference between media， incubation temperatures and culture times on the choice of hemodialysate and reverse osmosis water for microbiological monitoring of heterotrophic plate count， and further to confirm the best culture method of microbiological monitoring for hemodialysate and reverse osmosis. MethodsThe polluted samples were simulated by using sterile dialysate and sterile reverse osmosis water， and then were incubated in 25℃ and 35℃ with eosin methylene blue agar(EMB)， tryptone soy agar(TSA) and tryptone glucose extract agar(TGEA)for 48 h， 72 h and 7 d， respectively. ResultsEither on 25℃ or 35℃， the bacteria′s growth condition in TGEA was better than those under other conditions(P<0.05). The colony numbers of 72 h incubation were more than those of 48 h incubation in all media(P<0.05). However， there was no statistical significance of colony numbers between 7 d incubation and 72 h incubation(P > 0.05). ConclusionsThe microbiological monitoring for hemodialysate and reverse osmosis water needs to use pour plate method with TGEA， incubating in 25℃ and 35℃ for 72 h， which may provide the effective reports for ensuring the safety of hemodialysis patients.

Key words:Hemodialysate; Reverse osmosis water; Heterotrophic plate count

血液透析主要是靠血液与透析液在半透膜间进行弥散作用移除尿毒素，水是很好的溶剂，但进行高通量血液透析时微生物及其代谢产物会通过反弥散作用(back filtration)或透析器破膜方式进入血液，特别是当以透析液做为补充液时，微生物及其代谢产物更会直接进入人体[1-2]，导致菌血症和脓毒血症的发生。因此，要避免血液透析带来的长期慢性微发炎反应，让透析更安全，透析液的微生物监测是血液透析质量达标的第一步[3]。……