Yang J L, Yang Z Y, Luo G X, Wang X R, Li J L, Yang C X
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 1989;7(3):174-6.
A simple and rapid method for detecting Plasmodium falciparum in human blood was used in this study. The assay is based on DNA-DNA spot hybridization. For this purpose, the total genomic DNA of P. falciparum was isolated and purified from the parasite cultured in vitro. Then the total genomic DNA was used as a probe and labelled with [alpha-32P]-dATP by nick-translation. Twenty-five test samples, ten-microliter lysed infected blood each, were spotted onto dry nitrocellulose paper and hybridized with labelled genomic DNA. After hybridization, the paper was exposed to X-ray film for autoradiography, resulting in an image in places where hybridization occurred. The result shows that the assay appears to be sensitive enough to detect parasitaemia up to 0.0009%. No visible hybridization was detected in normal human blood or human leukocytes. Many samples can be processed simultaneously. This may be applicable to mass survey for detecting P. falciparum in epidemiological study (Fig. 2).
本研究采用一种简单快速的方法检测人体血液中的恶性疟原虫。该检测方法基于DNA-DNA斑点杂交。为此，从体外培养的疟原虫中分离并纯化出恶性疟原虫的总基因组DNA。然后将总基因组DNA用作探针，通过切口平移法用[α-32P]-dATP进行标记。将25个检测样品（每个样品为10微升裂解的感染血液）点样到干燥的硝酸纤维素纸上，并与标记的基因组DNA杂交。杂交后，将纸暴露于X射线胶片进行放射自显影，在发生杂交的部位产生图像。结果表明，该检测方法似乎足够灵敏，能够检测到低至0.0009%的疟原虫血症。在正常人血液或人白细胞中未检测到可见的杂交信号。可以同时处理多个样品。这可能适用于流行病学研究中检测恶性疟原虫的大规模调查（图2）。
