基于SSU rRNA基因的巢氏PCR隐孢子虫系统发育研究  

Molecular and phylogenetic characterization of Cryptosporidium by nested PCR based on SSU rRNA gene

作　　者：张晓[1] 吕静婷 杨璐璐[1] 孙宗科[1] 王友斌[1] 丁培[1] 武利平[1] 丁珵[1] 毛怡心 李霞[1] ZHANG Xiao;Lü Jing-ting;YANG Lu-lu;SUN Zong-ke;WANG You-bin;DING Pei;WU Li-ping;DING Cheng;MAO Yi-xin;LI Xia(Naliomd histitute of Environmental Health,Chinese Center for Disease Control and Prevention,Beijing 100021,China;不详)

机构地区：[1]中国疾病预防控制中心环境与健康相关产品安全所,北京100021 [2]北京市东城区卫生局卫生监督所

出　　处：《环境与健康杂志》2019年第6期511-514,F0003,共5页Journal of Environment and Health

基　　金：卫生部卫生行业专项(201302004).

摘　　要：目的鉴定由安徽某地鸭粪便中分离到的隐孢子虫虫种,并完善隐孢子虫DNA提取方法。方法对比液氮反复冻融提取DNA方法及普通DNA提取方法。对由某地区鸭粪便中分离到的隐孢子虫提取DNA,并对其SSU rRNA基因进行巢式PCR扩增,通过序列比对分析和构建系统进化树,解析隐孢子虫的种属关系。结果液氮反复冻融法更易于提取隐孢子虫DNA。该地区鸭粪便感染的隐孢子虫为贝氏隐孢子虫(Cryptosporidium baileyi,C.baileyi)。结论液氮反复冻融提取DNA方法对于检测被隐孢子虫感染的动物粪便切实有效。该地区的鸭存在贝氏隐孢子虫感染,需引起养殖者的重视。Objective To identify the Cryptosporidium species and genotypes in the duck feces from Anhui,and improve the DNA extraction method of Cryptosporidium. Methods Comparison of repeated freezing-thawing in liquid nitrogen and common DNA extraction methods was conducted. The DNA of Cryptosporidium isolated from duck feces was extracted and tested by nested PCR amplification of the 18 S rRNA gene. Results The application of repeated freezing-thawing in liquid nitrogen could make it easier to extract Cryptosporidium DNA. The Cryptosporidium isolates were C. baileyi by this method. Conclusion The method of extracting DNA by repeated freezing-thawing in liquid nitrogen is effective for detecting Cryptosporidium-infected animal feces. The ducks in this area present the infection with C. baileyi and the attention should be paid to.

关 键 词：鸭 隐孢子虫 巢式PCR SUU RRNA 

分 类 号：R117[医药卫生—公共卫生与预防医学]