摘要
[目的 ]建立适合于疟疾流行区现场应用的复式 PCR检测系统。 [方法 ]采用生理盐水处理样本 ,设计针对恶性疟原虫 (P.f) p BPK1- 14基因和间日疟原虫 (P.v)线粒体细胞色素 C氧化酶基因 CO 合成引物 ,混合一次扩增检测P.f 和 P.v。[结果 ]P.f 及 P.v模板分别被扩增出一条 2 10 bp和 370 bp的片段 ,与食蟹猴疟原虫 (P.c)、鼠伯氏疟原虫(P.b)及正常人白细胞无交叉 ,且可检测不同地理株的 P.f 和 P.v,P.f 和 P.v的敏感度分别为 5 .5× 10 - 7和 1.5×10 - 6 ,检测 146份疟区发热病人 ,与镜检结果相比 ,其敏感性和特异性 P.f为 97.9%和 98.7% ,P.v为 10 0 %和 98.7%。[结论 ]该复式 PCR检测系统适应于现场疟疾流行病学监测 ,防治效果考核 ,疑似疟疾病人的诊断及虫种鉴定。
Objective] To study the multiplex PCR for suitable to detect Plasmodium in field. [Methods] The samples were treated with Normal saline (N.S),malaria parasites DNA were amplified by the multiplex PCR using two sets of primers from the P.f moderately repetitive DNA sequence pRBK 1 14 and Pv COIII gene . [Results] A 210 bp product for P.f and 370 bp product for P.v were amplified by multiplex PCR, no special products were amplified for P.c, P.b and human WBC. P.f and P.v samples from different areas show the same amplified results, the method can detect parasitemia level as low as 5.5×10 -7 for P.f and 1. 5×10 -6 for P.v. A total of 146 fever samples were detected,the sensitivity and specificity were 100% and 98 7% for P.v, 97 9% and 98.7% for P.f. [Conclusion] The multiplex PCR can be suitable for The field malaria epidemiological surveillance, evaluation of the control effect ,diagnosis and determination of malaria species.
出处
《海峡预防医学杂志》
CAS
2001年第1期7-10,共4页
Strait Journal of Preventive Medicine
关键词
复式PCR
疟原虫
检测
应用
疟疾
诊断
multiplex polymerase chain reaction
plasmodium
detection
