聚合酶链反应检测实验动物弓形虫核酸的研究  

Study on Detecting Nucleotide of Toxoplasmosis in Laboratorium Animals with Polymerase Chain Reaction

作　　者：丛斌[1] 姚玉霞[1] 尤红煜[1] 彭郁葱[1] 

机构地区：[1]河北医科大学实验动物学部分子生物学研究室

出　　处：《中国实验动物学报》1998年第2期9-12,共4页Acta Laboratorium Animalis Scientia Sinica

摘　　要：建立弓形虫动物模型，常规方法提取肝、脾、肾、肺等组织DNA，应用聚合酶链反应（PCR）扩增，产物经电泳检测显示199bp的弓形虫特异带谱。并以γ－32p标记克隆的弓形虫特异DNA片段为探针，对扩增产物行Southern印迹分析，结果上述4种标本均出现阳性杂交带，进一步证实扩增条带是弓形虫特异DNA顺序。同时用酶标法检测显示鼠血清弓形虫抗体IgG；阳性。组织病理学检查结果，肝组织损伤较严重，肝细胞肿大，肝窦消失，脾、肾、肺组织可见轻微的病理改变。另外本文介绍一种简单PCR方法[1]，取鼠尾静脉血2μl直接进行扩增，结果与酚-氯仿法提取的DNA扩增结果一致。The DNAs were extracted from the liver, spleen, kidney and lung tissue of the animalmodels (KM mice) that were infected with Toxoplasma gondii by routine methods and the Toxoplasma gondii specific amplified bands (199bp) were shown in the PCR products from DNAs.Using r-32p-cloned T. gondii specific DNA fragment as probe, the result of southern blot assayshown that the probe only hybridized to the specific amplified DNA bands from the DNAs ofabove organs, which further proved that specific amplified bands were T. gondii specific DNA scquence. Also the T. gondii antibody(IgG) was POsitive in the serum of the KM mice with enzymelinked immunosorbent assay and the result of histopathology shown that lived tissue was injuredseriously but the lesion of spleen, kidney and lung tissue were slight. Additionally this article introduced a simple PCR method that 2μl blood from the m'ice caudal vein was directly amplifed andthe result was the same as that of DNA amplification with phenol-chloroform extractionmethod.

关 键 词：弓形虫抗体 扩增产物 阳性 肝组织损伤 肝窦 酶标法 严重 聚合酶链反应检测 杂交 酚-氯仿法 

分 类 号：Q78[生物学—分子生物学] R382.5[医药卫生—医学寄生虫学]