摘要
【目的】建立特异PCR方法诊断猪弓形虫病。【方法】以核糖体DNA第一内转录间隔区(ITS1)序列作为弓形虫种特异的遗传标记,优化PCR反应条件,对猪弓形虫分离株和临床病料进行检测。随机取阳性PCR扩增片段进行克隆,测序鉴定。【结果】在猪弓形虫分离株中PCR扩增出目的条带,且在肺门淋巴结和肠系膜淋巴结中扩增出特异性条带;PCR方法能检测到的最低DNA量为7.81 pg/μL;测序结果显示核苷酸序列与Genbank中已登录的猪弓形虫IST1基因序列相应部分序列完全相同。【结论】该PCR方法具有较高的敏感性和特异性,是诊断猪弓形虫病的一种快速检测方法。
【Objective】To establish specific PCR assay for diagnosing swine Toxoplasmosis.【Method】The first internal transcribed spacer sequence（IST1） of ribosomal DNA was used as the specific genetic marker for Toxoplasma gondii,the PCR reaction condition was optimized,the isolated strain and clinical infected materials of swine Toxoplasma were tested.Positive PCR amplified fragment was cloned and sequenced.【Result】DNA specific fragment for T.gondii strain was amplified,and specific bands were amplified in the bronchopulmonary lymph node and mesenteric lymph node ;the assay was able to detect as little as 7.81 pg/uL DNA of T.gondii;the nucleotide sequence was completely the same to the published homologous IST1 gene sequence of swine T.gondii in Genbank.【Conclusion】The PCR assay was highly sensitive and specific,which is a rapid method for diagnosing swine toxoplasmosis.
出处
《新疆农业科学》
CAS
CSCD
北大核心
2010年第9期1823-1827,共5页
Xinjiang Agricultural Sciences
基金
新疆生产建设兵团农业科技攻关计划项目(2007GG06)
国家绒毛用羊产业技术体系建设专项资金(NYCYTX-40-13)
关键词
猪
弓形虫
PCR
诊断
swine
Toxoplasma gondii
PCR
diagnosing
