用^(32)P标记海南株基因组DNA探针检测我国恶性疟原虫

Detection of Plasm odium falciparum From China with Genomic DNA Probe of Hainan Isolates

作者从人工培养的海南株恶性疟原虫提取基因组DNA,以^(32)P-dATP用切口移位法标记,采用斑点杂交法检测培养的海南株、云南株及女徽株恶性疟原虫及其提取的DNA.结果表明,该探针可检测出各株疟原虫的密度和DNA水平如下:海南株为0.0001％和1pg众两株均为0.001％和10pg.诊断海南省和云南省恶性疟患者的血标本共39份,其中38份阳性.在10份间日疟患者血标本中7份出现交叉反应.此探针与约氏疟原虫、食蟹猴疟原虫和诺氏疟原也呈现交叉反应,但不与正常人血细胞发生杂交反应。

The total genomic DNA was isolated from cultured P. falciparum Hainan Feel HN and was labelled with α - 32P-dATP through nick - translation. The gcnomic DNA probe was hybridized with DNA of P. falciparum Hainan, Yunnan and Anhui isolates. It was allowed to detect 0.0001% P. falciparum Hainan parasitemia and 0.001% Yunnan and Anhui parasitcmia in 20μ1 of blood samplc. 1 pg DNA of P. falcipwum Hainan and 10 pg DNA of Yunnan and Anhui isolates were detected The probe was used for detection of P. falciparum in 39 malarial patient samples from Hainan and Yunnan and the hybridization assay had a sensitivity of 97.4% (38/39). It was cross-hybridized with DNA of P. vivax, P. knowlcsi, P. cynomolgi, and P. yoclii, but not with human DNA.