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基于巢式PCR扩增恶性疟原虫特异性小亚基核糖体DNA片段的疟疾诊断研究

[Studies on diagnosis of falciparum malaria based on amplifying specific SSUrDNA fragment with nested PCR].

作者信息

Wan L, Chen P, Xue C, Jiang S

机构信息

Department of Parasitology, Fourth Military Medical University, Xi' an.

出版信息

Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 1995;13(3):174-7.

PMID:8556790
Abstract

Two pairs of primers specific to small subunit ribosomal DNA of Plasmodium falciparum were designed and the expected SSUrDNA fragment was amplified for detecting P. falciparum infection with double-temperature-nested polymerase chain reaction using DNA prepared by boiling method. The results showed that the nested PCR could amplify a constant size of desired SSUrDNA fragment of P. falciparum which was further confirmed by digestion of restriction endonuclease and could detect parasitemia level of 0.8 x 10(-6). It has great potentials for identifying Plasmodium species in ring form of erythrocytic stage and detecting mixed Plasmodium infections. Therefore, it is suggested that this method is sensitive, accurate, simple and rapid in detecting Plasmodium falciparum in blood samples for malaria diagnosis.

摘要

设计了两对针对恶性疟原虫小亚基核糖体DNA的引物,并使用煮沸法制备的DNA,通过双温嵌套聚合酶链反应扩增预期的小亚基核糖体DNA片段,以检测恶性疟原虫感染。结果表明,嵌套式PCR能够扩增出恒定大小的恶性疟原虫所需小亚基核糖体DNA片段,经限制性内切酶消化进一步证实,且能检测到0.8×10⁻⁶的原虫血症水平。该方法在鉴定红细胞期环状体疟原虫种类和检测混合疟原虫感染方面具有很大潜力。因此,建议该方法在检测血样中的恶性疟原虫以进行疟疾诊断时灵敏、准确、简便且快速。

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