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应用氯化锶对小鼠卵母细胞孤雌活化的研究
引用本文:杨红,陈建泉,刘海峰,李厚达,顾惠心,成国祥.应用氯化锶对小鼠卵母细胞孤雌活化的研究[J].实验动物与比较医学,2003,23(1):15-17.
作者姓名:杨红  陈建泉  刘海峰  李厚达  顾惠心  成国祥
作者单位:上海市实验动物质量监督检验站,200032
基金项目:上海市科技发展基金资助项目 ( 0 0 49190 71)
摘    要:根据弓形虫P30基因序列设计二对引物分别进行了PCR一次扩增和套式扩增,结果这两对引物在一次扩增和套式扩增中分别出现预期的扩增片段914bp,522bp和522bp;套式扩增出现的条带比一产供销扩增出现的条带亮度明显增强,提示套式扩增比一次扩增具有更高的敏感性,用外引物进行扩增后,最低可以检测到1pg的弓形虫DNA,说明所建立的反应体系具有高度的敏感性。用内引物对人工感染弓形虫的ICR小鼠血液,组织进行弓形虫DNA的检测,结果均扩增出522bp的扩增带。根据B1基因序列设计的弓物进行PCR一次扩增和半套式扩增。结果均出现预期的扩增片段619bp,362bp和362bp;用半套式扩增检测感染弓形虫形虫的ICR小鼠血液,组织时可扩增出522bp的619bp,362bp和362bp;用半套式扩增检测感染弓形虫的ICR小鼠血液,组织时可扩增出522bp的扩增带,半套式扩增比一次扩增更敏感。

关 键 词:检测方法  弓形虫  一次PCR  套式PCR  半套式PCR  ICR小鼠
文章编号:1004-8448(2003)01-0015-04
修稿时间:2002/2/27 0:00:00

Study on the Induction of Parthenogenetic Activation in Mouse Oocytes by Strontium Chloride
YANG Hong,CHEN Jian-quan,LIU Hai-feng,LI Hou-d,GU Hui-xin,CHENG Guo-xiang.Study on the Induction of Parthenogenetic Activation in Mouse Oocytes by Strontium Chloride[J].Laboratory Animal and Comparative Medicine,2003,23(1):15-17.
Authors:YANG Hong  CHEN Jian-quan  LIU Hai-feng  LI Hou-d  GU Hui-xin  CHENG Guo-xiang
Abstract:Two kinds of specific and sensitive methods for detecting Toxoplasma gondii was established by means of NT-PCR and One-Tube Hemi-Nested PCR. The DNAs were extracted from Toxoplasma gondii?Giardia lamblia and also blood?brain?liver?spleen?lung?kidney tissue of ICR mice infected with Toxoplasma gondii by routine methods. Two DNA fragments of the expected size (522 bp and 362 bp) was ampliflied from Toxoplasma gondii?blood?brain?liver?spleen?lung and kidney tissue by using NT-PCR and Hemi-Nested PCR rspectivelly,but no band was observed in Giardia lamblia. The sensitivity of the reaction was determined with different concentrations of genomic Toxoplasma gondii DNA, the DNA even less then 1pg could be detected by using single-step PCR. The NT-PCR or One-Tube Hemi-Nested PCR was more sensitive than single-step PCR.
Keywords:Toxoplasma gondii  Single-step PCR  NT-PCR  One-Tube Hemi-Nested PCR  ICR mice
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