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用于诊断恶性疟原虫疟疾的特异性DNA探针。

Specific DNA probe for the diagnosis of Plasmodium falciparum malaria.

作者信息

Barker R H, Suebsaeng L, Rooney W, Alecrim G C, Dourado H V, Wirth D F

出版信息

Science. 1986 Mar 21;231(4744):1434-6. doi: 10.1126/science.3513309.

Abstract

Malaria can be diagnosed either by direct microscopic examination of blood smears, which is time consuming and requires expertise, or by immunological techniques, which are effective but do not distinguish between past and present infections. In this study, a simple procedure was developed for spotting lysed blood from infected patients directly onto nitrocellulose paper and identifying the malaria species on the basis of hybridization of parasite DNA with a species-specific probe. A genomic DNA library of Plasmodium falciparum was screened to detect clones containing DNA sequences that are highly repeated within the parasite genome. Several such clones were further analyzed to identify those that hybridize specifically with P. falciparum DNA but not with DNA from humans, P. vivax, or P. cynomolgi. This technique appears to be sensitive enough to detect 10 picograms of purified P. falciparum DNA (equivalent to 100 parasites) and in field studies is able to detect approximately 40 parasites per microliter of blood.

摘要

疟疾的诊断既可以通过对血涂片进行直接显微镜检查,这种方法耗时且需要专业知识,也可以通过免疫技术,免疫技术虽有效但无法区分既往感染和当前感染。在本研究中,开发了一种简单的程序,即将感染患者的溶血直接点样在硝酸纤维素纸上,并基于寄生虫DNA与物种特异性探针的杂交来鉴定疟原虫种类。对恶性疟原虫的基因组DNA文库进行筛选,以检测含有在寄生虫基因组中高度重复的DNA序列的克隆。对几个这样的克隆进行进一步分析,以鉴定那些与恶性疟原虫DNA特异性杂交但不与人类、间日疟原虫或食蟹猴疟原虫的DNA杂交的克隆。该技术似乎足够灵敏,能够检测到10皮克纯化的恶性疟原虫DNA(相当于100个疟原虫),并且在现场研究中能够检测到每微升血液中约40个疟原虫。

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