用PCR方法检测锥虫感染  

The detection of trypanosoma infection by PCR method

作　　者：陈小光[1] 陈汉妹 黄小敏 CHEN Xiao-guang;CHEN Han-mei;HUANG Xiao-min(Guangzhou Blood Center,Guangzhou,Guangdong 510095,China)

机构地区：[1]广州血液中心

出　　处：《中国国境卫生检疫杂志》2019年第6期428-429,449,共3页Chinese Journal of Frontier Health and Quarantine

摘　　要：目的 建立检测锥虫感染的PCR方法。方法 针对布氏锥虫表达位点相关基因(ESAG)设计引物并建立PCR扩增体系,检测其扩增效果。用梯度稀释的布氏锥虫DNA(10μg/μl、1 ng/μl、100 pg/μl、10 pg/μl、1 pg/μl和0.1 pg/μl)为模板评价其敏感性;用建立的PCR方法分别检测布氏锥虫、伊氏锥虫及克氏锥虫,评价方法的特异性,并比较PCR与镜检法的准确性。结果 PCR法扩增布氏锥虫的最低检测限为100 pg/μl;准确性为96.67%,高于镜检法的80.00%,差异有统计学意义(P <0.05)。建立的PCR方法检测10株布氏锥虫均为阳性,10株克氏锥虫均为阴性,1株伊氏锥虫为阳性。结论 PCR法检测锥虫感染具有较高的敏感性和特异性,在布氏锥虫感染的早期诊断中具有较高应用价值。Objective To develop a PCR method for the detection of trypanosomiasis infection.Methods The specific primer pair was designed targeted to the expression site-associated gene(ESAG)of Trypanosoma brucei,and PCR amplification system was established.The amplification effect was test.The sensitivity was assessed by using gradient concentrations of Trypanosoma brucei DNA template(10μg/μl,1 ng/μl,100 pg/μl,10 pg/μl,1 pg/μl and 0.1 pg/μl).The specificity was evaluated through the amplification of Trypanosoma brucei,Trypanosoma evansi and Trypanosoma cruzi.The accuracy of the PCR method and microscopy examination was compared.Results The detect limitation of the PCR method developed was 100 pg/μl.The accuracy was 96.67%,which was higher than that of microscopy examination(80.00%),the difference was statistically significant(P<0.05).All of 10 strains of Trypanosoma brucei had been detected positive by using this PCR method,but 10 strains of Trypanosoma cruzi were all negative,and 1 strain of Trypanosoma evansi was positive.Conclusion The PCR showed high sensitivity and specificity in the detection of Trypanosoma brucei infection,with high application value in the early diagnosis of trypanosomiasis.

关 键 词：聚合酶链式反应 锥虫 布氏锥虫 锥虫病 

分 类 号：R382.9[医药卫生—医学寄生虫学]