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应用聚合酶链反应/DNA探针鉴定中国间日疟原虫环子孢子蛋白基因型

Use of PCR/DNA probes to identify circumsporozoite genotype of Plasmodium vivax in China.

作者信息

Han G D, Zhang X J, Zhang H H, Chen X X, Huang B C

机构信息

Shandong Institute of Parasitic Diseases, Jining, China.

出版信息

Southeast Asian J Trop Med Public Health. 1999 Mar;30(1):20-3.

Abstract

The paper reports the result of identifying cirumsporozoite (CS) genotype of Plasmodium vivax by using PCR/DNA probe labeled with biotin. The sensitivity of this method to detect patient blood samples was 0.2 parasite/microl and also with high specific to P. vivax. CS genes from 52 blood samples collected from patients with P. vivax in Hainan and Yunnan Provinces were amplified by PCR and 49 were positive by gel-e electrophoresis analysis, positive rate was 94%. Then the amplified CS genes further were probed with special oligoprobes (PV210 and PV247) that hybridized with the predominant CS repeat region and the variant CS repeat region. The results showed 46 (88.5%) PV210 positive and 6 (11.5%) PV247 positive; 2 hybridized with both probes. The variant genotype was present only in samples from Yunnan Province. The above results showed that the PCR/DNA probe labeled with biotin was highly sensitive and specific to P. vivax and found a CS variant genotype of P. vivax in Yunnan Province of China.

摘要

本文报道了使用生物素标记的PCR/DNA探针鉴定间日疟原虫环子孢子(CS)基因型的结果。该方法检测患者血样的灵敏度为0.2个寄生虫/微升,对间日疟原虫也具有高度特异性。对从海南和云南省间日疟患者采集的52份血样中的CS基因进行PCR扩增,经凝胶电泳分析49份呈阳性,阳性率为94%。然后用与主要CS重复区域和变异CS重复区域杂交的特殊寡核苷酸探针(PV210和PV247)对扩增的CS基因进行检测。结果显示,46份(88.5%)PV210阳性,6份(11.5%)PV247阳性;2份与两种探针均杂交。变异基因型仅存在于云南省的样本中。上述结果表明,生物素标记的PCR/DNA探针对间日疟原虫具有高度敏感性和特异性,并在中国云南省发现了间日疟原虫的一种CS变异基因型。

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