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弓形虫的冷冻保存及其在小鼠体内抗原性的研究。

Studies on cryopreservation of Toxoplasma gondii and its antigenicity in mice.

作者信息

Lin D B, Su K E, Yu J C

机构信息

School of Medical Technology, Chung Shan Medical and Dental College, Taichung, Taiwan, Republic of China.

出版信息

Am J Trop Med Hyg. 1995 Oct;53(4):392-6. doi: 10.4269/ajtmh.1995.53.392.

Abstract

Two strains (RH and GC, the latter of which is a Taiwan isolate of porcine origin) of Toxoplasma gondii were kept at -20 degrees C, -60 degrees C, and in liquid nitrogen (-196 degrees C) to follow the time course change in viability and virulence of the parasites by direct count and animal inoculation methods. Changes in antibody titers in some of the mice inoculated with the thawed organisms were assayed by the indirect immunofluorescent antibody test. Viability and virulence of T. gondii were best preserved by storage in liquid nitrogen. Tachyzoites kept in liquid nitrogen for eight years still can lead to the death of the injected mice in 2-3 weeks. Virulence of the tachyzoites could be maintained for eight weeks at most at -20 degrees C and -60 degrees C. Dimethylsulfoxide (DMSO) seemed to be a better cryoprotectant for T. gondii than glycerol, but the DMSO-preserved organisms resulted in fewer tachyzoite-containing peritoneal exudates in inoculated mice than the glycerol-preserved organisms. The local isolate (GC strain) tachyzoites tolerated cryopreservation less well than the RH strain parasites. Low antibody titers (at most 1:64) were produced in mice that survived more than 16 days after inoculation with thawed tachyzoites.

摘要

将两株刚地弓形虫(RH株和GC株,后者是源自猪的台湾分离株)分别保存在-20℃、-60℃和液氮(-196℃)中,通过直接计数和动物接种方法追踪寄生虫活力和毒力随时间的变化。对接种解冻后虫体的部分小鼠,采用间接免疫荧光抗体试验检测抗体滴度的变化。刚地弓形虫的活力和毒力通过液氮保存最佳。保存在液氮中的速殖子8年后仍能在2至3周内导致接种小鼠死亡。速殖子的毒力在-20℃和-60℃下最多可维持8周。二甲基亚砜(DMSO)对刚地弓形虫而言似乎是比甘油更好的冷冻保护剂,但与甘油保存的虫体相比,DMSO保存的虫体接种小鼠后含速殖子的腹腔渗出液较少。本地分离株(GC株)速殖子对冷冻保存的耐受性不如RH株寄生虫。接种解冻后速殖子存活超过16天的小鼠产生的抗体滴度较低(最高1:64)。

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