作者：车成业 作者单位：青岛大学医学院附属医院眼科,山东 青岛 266003
【摘要】 目的 了解6种角膜接触镜多功能护理液和加入甲硝唑滴眼液的护理液对自生生活性棘阿米巴原虫的杀伤效果。方法 将6种多功能护理液分别加入96孔培养板中，每种护理液占用48孔，其中24孔滴入棘阿米巴悬液，另外24孔先滴入甲硝唑滴眼液后再滴入棘阿米巴悬液，室温静置8 h后在倒置显微镜下观察棘阿米巴的形态变化和数量。将残存的棘阿米巴原虫分别在PYG培养液中培养5 d，观察其形态、活性与增殖能力的变化。结果 单纯护理液组1～6号棘阿米巴原虫检出率分别为0、8.3%、29.1%、41.7%、62.5%、79.2%，加入甲硝唑滴眼液后护理液1～6号棘阿米巴原虫检出率分别为0、0、4.2%、8.3%、16.7%、16.7%，3～6号护理液加与不加甲硝唑滴眼液杀伤棘阿米巴原虫的效果差异有统计学意义（χ2=3.75～18.78，P<0.05）。残存的棘阿米巴原虫经培养后活力与增殖力减弱。结论 部分多功能护理液对棘阿米巴原虫的杀伤效果不佳，添加甲硝唑滴眼液后杀伤效果明显提高。
【关键词】 棘阿米巴属 角膜接触镜多功能护理液 甲硝唑
THE KILLING EFFICIENCY OF ARILIN AND CONTACT LENS SOLUTIONS ON ACANTHAMOEBA
CHE CHENGYE, ZHAO GUIQIU, ZHANG JING, et al
The Department of Ophthalmology, The Affiliated Hospital of Qingdao University Medical College, Qingdao 266003, China
［ABSTRACT］ Objective To analyze the killing efficiency of six kinds of contact lens solutions and solutions with arilin on freeliving Acanthamoeba. Methods The solutions were added into 96well microtiter plates respectively. Each care solution used 48 holes,24 of those holes were added with suspension of Acanthamoeba. The other 24 holes were added in arilin and suspension of Acanthamoeba. After standing in room temperature for eight hours, the morphologic changes and quantity of the remnant Acanthamoeba were observed under an inverted microscope. The remnant Acanthamoeba were cultivanted in PYGculture medium for five days. Then, their variation of appearance, activity and reproductive activity were observed. Results In the six experimental groups using contact lens solutions only, the detection rates of Acanthamoeba of were 0,8.3%, 29.1%,41.7%,62.5% and 79.2%, respectively. After arilin was added, the detection rates of Acanthamoeba of those six groups were 0,0,4.2%,8.3%,16.7% and 16.7%, respectively. From group 3 to group 6, after added in arilin, the differences of the killing efficiency of contact lens solutions were signficant (χ2=3.75-18.78，P<0.05). Cultivation of the remnant Acanthamoeba showed a reduction in their activity and proliferative ability. Conclusion The killing efficiency of some contact lens solutions on freeliving Acanthamoeba is not satisfied. Arilin can improve the killing efficiency of contact lens solutions.
［KEY WORDS］ Acanthamoeba castellanii; Contact lens slutions; Arilin
棘阿米巴角膜炎由棘阿米巴原虫感染引起，是一种严重威胁视力的角膜炎。国外研究已证实，配戴被原虫污染的角膜接触镜及角膜轻度擦伤是诱发棘阿米巴角膜炎的主要原因[1]。合格的角膜接触镜多功能护理液应具备清洁、消毒和去蛋白三大功能[2]，所以理应在棘阿米巴角膜炎的预防中起到极为重要的作用。本实验选用市售的6种多功能护理液与自生生活性棘阿米巴原虫共同培养，以检验其杀伤效果，并与添加甲硝唑滴眼液的护理液的杀伤效果进行比较。
1 材料与方法
1.1 棘阿米巴原虫的培养
标准棘阿米巴菌种（CW1）购于北京市眼科研究所。实验前在28 ℃温箱，含有青霉素(500 kU/L) 和链霉素(500 kU/L)的蛋白胨酵母葡萄糖（PYG）培养液中无菌纯培养3～5 d至对数生长期。倒置显微镜下观察可见：PYG培养液中，阿米巴原虫有滋养体（80%以上）和包囊两种形态。滋养体多为不规则长圆形,体表有多个细小的棘刺状突起,胞质内可见多个空泡；包囊类圆形,有两层囊壁,内壁呈多面体形。血细胞计数板计数滋养体，制成滋养体浓度为3×107/L的悬液，振荡混匀。
1.2 多功能护理液
选择市售的6种角膜接触镜多功能护理液，均不包含甲硝唑及其有效成分。分别编号为1、2、3、4、5、6号。生产日期均为2007年4～8月，保质期2年。实验日期均在保质期内。
1.3 实验方法
使用3张无菌96孔板，每种护理液占用48孔，使用微量加样器在每个孔中加入护理液100 μL，再于其中的24孔中加入2 g/L甲硝唑滴眼液100 μL，所有孔中加入浓度为3×107/L的阿米巴原虫悬液1 μL。倒置显微镜观察，全部孔中均可见棘阿米巴原虫。室温静置8 h后在倒置显微镜下观察阿米巴原虫的形态变化和数量。
1.4 残存阿米巴原虫的培养
将3张96孔板中剩余的1～6号护理液收集后离心去上清（2 000 r/min，离心5 min），再加入PYG培养液2 mL进行培养。同时设置对照组，取浓度为3×107/L的阿米巴悬液24 μL，同样方法离心去上清，加入2 mL的PYG培养液培养。5 d后观察并比较两组残存阿米巴原虫的形态、活性与增殖能力的变化。
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