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目的探讨群组PCR策略高通量筛选不同密度疟原虫的敏感性和特异性,以期用于消除疟疾后的疟疾病例监测。方法采用2种敏感的PCR方法分别对8个不同密度梯度疟原虫下单个标本组,5个标本群组,25个标本群组,50个标本群组,100个群组进行敏感性和特异性检测,并对8个梯度密度疟原虫进行显微镜检查和免疫学诊断。结果群组PCR检测疟原虫的敏感性随着原虫密度梯度的下降和群组大小的增加呈现下降趋势。5个样品群组和25个样品群组均可对低密度疟原虫进行检测,采用核糖体基因检测法(18SSSu RNA基因)比细胞色素B基因(CYTB)的敏感度最高,最低可检测到0.8p/μl疟原虫DNA;5个样品群组检测的敏感性高于25个样品群组。结论在敏感性和特异性一定的情况下,群组PCR可对低密度疟原虫感染者进行高通量筛选,较逐一筛选节省大量费用,适合于在消除疟疾阶段对疟疾病例进行监测。
OBJECTIVE: To investigate the sensitivity and specificity of high-throughput screening of malaria parasite of different densities by group PCR strategy in order to monitor malaria cases after eliminating malaria. Methods Two sensitive PCR methods were used to detect the sensitivity and specificity of single samples, five specimens, 25 specimens, 50 specimens and 100 specimens of 8 different density gradient Plasmodium strains The microscopic examination and immunological diagnosis of 8 gradient density Plasmodium species were carried out. Results The sensitivity of group PCR for Plasmodium falciparum showed a decreasing trend with the decrease of protozoan density gradient and group size. Low-density Plasmodium could be detected in 5 sample groups and 25 sample groups, and the highest sensitivity was detected by ribosomal gene assay (18SSSu RNA gene) than cytochrome B gene (CYTB). The lowest detection limit was 0.8 p / μl Plasmodium DNA; the sensitivity of the five sample groups was higher than that of the 25 sample groups. Conclusion In the case of certain sensitivity and specificity, the group PCR can be used for high-throughput screening of low-density Plasmodium infection, saving a lot of costs compared with screening one by one, and is suitable for monitoring malaria cases in the phase of malaria elimination.