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用于检测间日疟原虫环子孢子蛋白多态性VK 247和VK 210的DNA探针的比较试验

Comparative test of DNA probes for detection of Plasmodium vivax circumsporozoite protein polymorphs VK 247 and VK 210.

作者信息

Sattabongkot J, Suwanabun N, Rongnoparut P, Wirtz R A, Kain K C, Rosenberg R

机构信息

Department of Entomology, US Army Medical Component, Bangkok, Thailand.

出版信息

J Infect Dis. 1994 Feb;169(2):464-6. doi: 10.1093/infdis/169.2.464.

Abstract

Oligonucleotide probes specific to the characteristic repeat sequences of two alleles of the circumsporozoite protein gene of Plasmodium vivax (VK 210 and VK 247) were selected, synthesized, and tested on matched blood and sporozoite DNA amplified by polymerase chain reaction from 182 cases naturally acquired in Thailand. Probe results were compared to those of circumsporozoite phenotype-specific ELISAs used to evaluate sporozoites from the same cases. There was a 96% agreement between probe results for blood and for sporozoites. Although there was also a nearly complete agreement between probe and ELISA results for cases producing only VK 210 or VK 247 sporozoites, the probes detected 45% more mixed infections than did the ELISAs when used to test specimens from western and southern Thailand; there was no discrepancy when mixed cases from Cambodia were tested. Examination of Southern blots from ambiguous mixed cases demonstrated the presence of both genes, suggesting suppression of VK 247 in some mixed cases to numbers below those detectable by the ELISA.

摘要

选择、合成了针对间日疟原虫环子孢子蛋白基因两个等位基因(VK 210和VK 247)特征性重复序列的寡核苷酸探针,并对从泰国自然感染的182例病例中通过聚合酶链反应扩增的配对血液和子孢子DNA进行了检测。将探针检测结果与用于评估同一病例子孢子的环子孢子表型特异性酶联免疫吸附测定(ELISA)结果进行比较。血液和子孢子的探针检测结果有96%的一致性。虽然对于仅产生VK 210或VK 247子孢子的病例,探针和ELISA结果也几乎完全一致,但当用于检测泰国西部和南部的标本时,探针检测到的混合感染比ELISA多45%;检测来自柬埔寨的混合病例时没有差异。对不明确的混合病例的Southern印迹进行检查,证实了两个基因的存在,这表明在一些混合病例中VK 247被抑制到ELISA检测不到的数量以下。

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