中国血吸虫病防治杂志 ›› 2022, Vol. 34 ›› Issue (4): 400-.

• 实验研究 • 上一篇    下一篇

曼氏裂头蚴病动物模型的建立及诊治技术研究Ⅱ原尾蚴灌胃法建立曼氏裂头蚴病小鼠模型

汤宪时,许永良,王强,张英,仝德胜*   

  1. 国家卫生健康委员会寄生虫病预防和控制技术重点实验室、江苏省寄生虫与媒介控制技术重点实验室、江苏省血吸虫病防治研究所(江苏 无锡214064)
  • 出版日期:2022-08-31 发布日期:2022-08-31
  • 作者简介:汤宪时,男,硕士,助理研究员。研究方向:寄生虫分子生物学
  • 基金资助:
    江苏省卫生健康委科研项目(H2018099)

Establishment of an animal model of Sparganum mansoni infection and study on therapeutic methods II Establishment of a mouse model of sparganosis mansoni via oral administration of procercoids

TANG Xian⁃shi, XU Yong⁃liang, WANG Qiang, ZHANG Ying, TONG De⁃sheng*   

  1. Key Laboratory of National Health Commission on Parasitic Disease Control and Prevention; Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology; Jiangsu Institute of Parasitic Diseases, Wuxi, Jiangsu 214064, China
  • Online:2022-08-31 Published:2022-08-31

摘要: 目的 探索通过感染原尾蚴的剑水蚤灌胃小鼠建立曼氏裂头蚴病小鼠动物模型的可行性。方法 实验室条件下用曼氏裂头蚴感染家猫,收集当日新产猫粪,用去氯水淘洗后过滤获得曼氏迭宫绦虫虫卵,制成虫卵悬浊液。将20只C57BL/6j小鼠随机分为实验组和对照组,实验组15只、对照组5只。以曼氏迭宫绦虫钩球蚴感染野生剑水蚤,使每只剑水蚤体内含有3 ~ 5条原尾蚴;再将感染原尾蚴的剑水蚤灌胃实验组小鼠,每只小鼠灌入剑水蚤15只,对照组灌胃等体积去氯水。6个月后处死小鼠,分离疑似裂头蚴虫体,并用间接酶联免疫吸附试验检测小鼠血清中抗曼氏裂头蚴特异性IgG抗体水平。提取疑似虫体基因组DNA,以曼氏裂头蚴细胞色素C氧化酶1(CO1)基因片段为目的序列进行PCR扩增以鉴别疑似虫体。结果 实验组15只小鼠中,6只小鼠血清抗曼氏裂头蚴特异性IgG抗体检测呈阳性;于其中4只小鼠皮下发现并分离出乳白色虫体。每只小鼠均仅检出1条虫体,虫体形态与曼氏裂头蚴相似。基于CO1基因的疑似虫体PCR扩增产物在毛细管电泳上于151 bp处显示特异性条带,测序结果与已知曼氏裂头蚴序列100%符合。结论 本研究成功建立了一种曼氏裂头蚴病小鼠动物模型。

关键词: 曼氏裂头蚴病, 曼氏裂头蚴, 原尾蚴, 动物模型

Abstract: Objective To establish an animal model of sparganosis mansoni through oral administration of Cyclops infected with procercoids. Methods Domestic cats were infected with Sparganum mansoni under laboratory conditions, and fresh cat stool samples were collected, washed in dechlorinated water, and filtered. Spirometra mansoni eggs were collected and prepared into suspensions. Twenty C57BL/6j mice were randomly divided into the experimental group (n = 15) and the control group (n = 5). Wild Cyclops were infected with Spirometra mansoni coracidia to allow 3 to 5 procercoids in each Cyclop. Then, each mouse in the experimental group was given 15 Cyclops infected with procercoids by gavage, while mice in the control group were orally administered with the same volume of dechlorinated water. All mice were sacrificed after 5 months, and dissected, and suspicious Sparganum mansoni worms were collected. The serum specific IgG antibody against Sparganum mansoni was measured in mice using enzyme⁃linked immunosorbent assay (ELISA). Genomic DNA was isolated from suspicious Sparganum mansoni worms, and the specific Sparganum mansoni cytochrome oxidase I (COI) gene was amplified using PCR assay. Results Among the 15 mice in the experimental group, six were positive for the serum specific IgG antibody against Sparganum mansoni, and milky white worms were found and collected from the subcutaneous regions of 4 out of 6 mice. Only one worm was detected in each mouse, and the worm morphology was similar to Sparganum mansoni. Capillary electrophoresis of the PCR amplification products of COI gene presented a specific band with 151 bp in size, and sequencing analysis revealed 100% homology with Sparganum mansoni. Conclusions A mouse model of sparganosis mansoni is successfully created through oral administration of Cyclops infected with Spirometra mansoni procercoids.

Key words: Sparganosis mansoni, Sparganum mansoni, Procercoid, Animal model

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