[应用多重聚合酶链反应检测血样中恶性疟原虫和间日疟原虫的研究]

[Studies on detection of Plasmodium falciparum and Plasmodium vivax in blood samples by multiplex polymerase chain reaction].

作者信息

Chen S, Lu H, Gao Q, Tang X

机构信息

Department of Parasitology, Suzhou Medical College, Suzhou 215007.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 1999;17(6):380-3.

PMID:12563743

Abstract

AIM

To establish a sensitive and specific PCR-based method to detect Plasmodium falciparum and P. vivax in blood samples in a single amplification reaction.

METHODS

Malaria parasite DNA in blood was amplified by the multiplex polymerase chain reaction using two sets of primers derived from the P. f. moderately-repetitive DNA sequence and COIII gene of P.v.

RESULTS

A 206-bp product for P. f. and a 370-bp product for P.v. were amplified by multiplex PCR, being able to detect parasitemia level as low as 5 x 10(-7) for P. f. and 1.02 x 10(-6) for P. v. and having no cross-reaction with human leucocyte DNA. A total of 783 blood samples on the filter paper collected from patients attending to malaria clinics in malaria endemic areas were detected. The positive rate of multiplex PCR was 85.8%, the misdiagnosis rate was 0, and the under-diagnosis rate was 0.1%, while these three rates of microscopic examination were 84.9%, 3.1% and 1.0%, respectively. The concordance between the two methods was 95.8%.

CONCLUSION

The multiplex PCR method made the malaria detection more sensitive and specific than the microscopic examination and should be suitable for the diagnosis of malaria in mixed endemic areas, large-scale epidemiological studies, follow-up of drug treatment and donor blood screening.

摘要

目的

建立一种灵敏且特异的基于聚合酶链反应（PCR）的方法，以在单一扩增反应中检测血样中的恶性疟原虫和间日疟原虫。

方法

使用源自恶性疟原虫中度重复DNA序列和间日疟原虫细胞色素c氧化酶III（COIII）基因的两组引物，通过多重聚合酶链反应扩增血液中的疟原虫DNA。

结果

多重PCR扩增出了一条206bp的恶性疟原虫产物和一条370bp的间日疟原虫产物，能够检测到低至5×10⁻⁷的恶性疟原虫血症水平和1.02×10⁻⁶的间日疟原虫血症水平，且与人白细胞DNA无交叉反应。对从疟疾流行地区疟疾诊所就诊患者采集的783份滤纸片血样进行了检测。多重PCR的阳性率为85.8%，误诊率为0，漏诊率为0.1%，而显微镜检查的这三个率分别为84.9%、3.1%和1.0%。两种方法的一致性为95.8%。