猪附红细胞体50S核糖体基因PCR检测方法的建立及应用

根据GenBank上最新发布的猪附红细胞体基因组序列(NC-015155)设计一对引物,并以吉林省延边地区猪附红细胞体基因组DNA为模板,建立猪附红细胞体50 S核糖体基因PCR诊断方法,通过特异性、敏感性及临床应用试验验证,快速准确的检测出猪附红细胞体.试验结果显示,建立的猪附红细胞体PCR诊断方法扩增片段大小为10...

Establishment and Application of PCR Assay for Eperythrozoon suis Based on 50S Ribosome Gene

In order to detect Eperythrozoon suis rapidly and accurately,a pair of primers were designed based on the latest genome sequence of E.suis（NC₀15155） in GenBank.The genomic DNA of E.suis in Yanbian Jilin province was taken as a template to establish a PCR diagnostic method of E.suis 50S ribosome,and then specific test,sensitivity test and clinical practice test were finished.The results show that,according to the established PCR diagnostic method,the length of the amplified fragment is 1 099 bp（accession ID: JN166805）.Meanwhile,the sequence of the gene shares 93% homology with Illinois in GenBank（NC₀15155）.With this method,several pathogens can’t be amplified,such as Escherichia coli,Streptococcus,Mycoplasma hyopneumoniae E.wenyonii and so on.The lowest detectable limit of the method is 19 fg/μL for DNA content in E.suis.Moreover,45 swine blood samples from Yanbian area were detected with the method and compared with Giemsa staining of blood smear.Results indicated that,the PCR diagnostic method is specific,sensitive and accurate,which is totally suitable for detection of E.suis.