CCK-8法在阴道毛滴虫体外增殖检测中的应用研究  

The Application of CCK-8 Method in the Detection of Trichomonas Vaginalis Proliferation in vitro

作　　者：关玮 杨树国 李健 李晓强[2] 赵燕清 谢祎婷 GUAN Wei;YANG Shu-guo;LI Jian;LI Xiao-qiang;ZHAO Yan-qing;XIE Yi-ting(Department of Parasitology,School of Basic Medicine,Taihe Hospital,Hubei University of Medicine,Shiyan,Hubei 442000,China;Department of Clinical Laboratory Center,Taihe Hospital,Hubei University of Medicine,Shiyan,Hubei 442000,China)

机构地区：[1]湖北医药学院基础医学院寄生虫学教研室,湖北十堰442000 [2]湖北医药学院附属太和医院检验部,湖北十堰442000

出　　处：《湖北医药学院学报》2021年第6期591-596,共6页Journal of Hubei University of Medicine

基　　金：湖北省教育厅科学技术研究项目(Q20192102)。

摘　　要：目的:探究CCK-8法能否用于阴道毛滴虫体外增殖的检测。方法:通过半浓度梯度稀释法设置阴道毛滴虫TV36株虫量梯度,用CCK-8法制备滴虫定量标线。再用不同浓度甲硝唑处理滴虫,检测在药物评估中CCK-8法和显微镜计数法对滴虫虫量和生长抑制率的一致性。同时检测TV-THS1g、TV21、TV35三株滴虫的CCK-8定量标线以及甲硝唑敏感度。比较检测滴虫增殖的4种方法的优缺点。结果:使用CCK-8法制备的4株滴虫虫量标线,在1~4 h均具有良好的线性关系。不同浓度甲硝唑处理TV36后,CCK-8法和显微镜计数法对滴虫数量(ICC=0.98,P<0.0001)和滴虫生长抑制率(ICC=0.919,P<0.0001)的检测均具有高度一致性。使用两种方法检测的甲硝唑对TV36、TV-THS1g、TV21、TV35的半抑制浓度值也很相近。结论:CCK-8法能够用于阴道毛滴虫体外增殖的检测,可为抗滴虫感染的药物筛选及其他增殖相关研究提供一种重要的检测工具。Objective To explore whether Cell Counting Kit-8(CCK-8)could be used for proliferation detection of Trichomonas vaginalis in vitro.Methods The worm population gradient of Trichomonas vaginalis TV36 strain was set by the half-concentration gradient dilution method,and the trichomoniasis quantitative marking line was prepared by the CCK-8 method.Then,different concentrations of metronidazole were used to treat Trichomonas,and the consistency of the CCK-8 method and the microscope counting method on the amount and growth inhibition rate of Trichomonas was detected in the drug evaluation.Meanwhile,the standard curves of other three T.vaginalis strains including TV-THS1g,TV21,TV35 and their sensitivity to metronidazole were detected by CCK-8.Finally,the advantages and disadvantages of four methods for detecting the proliferation of Trichomonas were compared.Results The standard curves for quantification of T.vaginalis(TV36,TV-THS1g,TV21 and TV35)generated by CCK-8 all have good linearity within 1~4 h.After treatment of TV36 with metronidazole in different concentrations,the cell counting results(ICC=0.98,P<0.0001)and the growth inhibition rates of T.vaginalis(ICC=0.919,P<0.0001)obtained by the two methods have excellent consistency.The half-inhibitory concentrations(IC_(50))of metronidazole on TV36,TV-THS1g,TV21,and TV35 detected by the two methods are also very similar.Conclusion The CCK-8 method can be used to detect the proliferation of Trichomonas vaginalis in vitro,and can provide an important detection tool for drug screening against trichomonas infection and other proliferation-related research.

关 键 词：阴道毛滴虫 CCK-8 显微镜计数 增殖检测方法 

分 类 号：R711.31[医药卫生—妇产科学]