基于血球计数板法的球虫卵囊计数方法研究  

Research on Coccidian Oocyst Counting Method Based on the Hemocytometer Counting Chamber Method

作　　者：李建[1] 辛凌翔[1] 王秀丽[1] 任小侠[1] 刘元杰 姚文生[1] 张一帜 魏津[1] 马欣[1] 王甲[1] 冯妍 赵浩然 刘燕[1] 朱良全[1] LI Jian;XIN Lingxiang;WANG Xiuli;REN Xiaoxia;LIU Yuanjie;YAO Wensheng;ZHANG Yizhi;WEI Jin;MA Xin;WANG Jia;FENG Yan;ZHAO Haoran;LIU Yan;ZHU Liangquan(China Institute of Veterinary Drug Control,Beijing 100081,China)

机构地区：[1]中国兽医药品监察所,北京海淀100081

出　　处：《中国兽医杂志》2025年第3期74-80,共7页Chinese Journal of Veterinary Medicine

基　　金：国家重点研发计划(2022YFD1800903);兽药行业公益性重点专项(GY202104)。

摘　　要：本试验旨在基于血球计数板法探究影响球虫卵囊计数准确性的因素,评价相关因素对结果准确性的影响程度,从而对目前广泛应用的球虫活疫苗卵囊计数方法进行改进。试验中对比不同放大倍数视野、不同生产厂家血球计数板、不同球虫孢子化卵囊浓度和助悬剂对球虫卵囊计数结果准确性的影响。结果显示,视野放大倍数、血球计数板、球虫孢子化卵囊浓度和助悬剂对球虫卵囊计数结果均有一定影响,其中,视野放大倍数有显著影响。添加助悬剂,调整样品球虫孢子化卵囊数量至181~343个/0.4μL,用改良Neubauer血球计数板在10×20倍显微镜下进行计数,结果较为准确,连续6次计数结果的变异系数基本控制在20%以内。结果表明,限定视野放大倍数、血球计数板和球虫孢子化卵囊浓度及添加助悬剂,可有效降低球虫卵囊计数结果的变异系数,提高计数结果的准确性,降低人员误差,为《中华人民共和国兽药典》改版修订提供了试验依据和理论参考。This study aims to explore the factors affecting the accuracy of coccidian oocyst counting using the hemocytometer counting chamber method,and to evaluate the impact of these factors on the result accuracy.The objective is to improve the currently widely used oocyst counting method for coccidian live vaccines.The experiment compared the effects of different magnification,different manufacturers'hemocytometer counting chamber,different oocyst concentrations of sporulated oocysts,and suspending agents on the accuracy of coccidian oocyst counting results.The results showed that the magnification,hemocytometer counting chamber,oocyst concentration,and suspending agents all had an impact on the counting accuracy,with magnification showing a significant effect.Adding a suspending agent and adjusting the oocyst concentration to 181-343 oocysts/0.4μL,and using an improved Neubauer hemocytometer counting chamber under a 10×20 magnification microscope yielded the most accurate results,with the coefficient of variation for six consecutive counts controlled within 20%.The results indicate that by controlling magnification,hemocytometer counting chamber type,oocyst concentration of sporulated oocysts,and adding a suspending agent,the coefficient of variation in oocyst counting results can be effectively reduced,improving the accuracy and minimizing human errors.These findings provide experimental evidence and theoretical reference for the revision and update of the Pharmacopoeia of the People's Republic of China.

关 键 词：卵囊计数 助悬剂 孢子化卵囊 球虫 血球计数板 

分 类 号：S855[农业科学—临床兽医学]