Ayyanathan K, Francis V S, Datta S, Padmanaban G
Astra Research Centre India, Bangalore, India.
Mol Cell Probes. 1995 Aug;9(4):239-46. doi: 10.1016/s0890-8508(95)90103-5.
The application of nucleic acid probes, in the detection of pathogenic micro-organisms, has become an integral part of diagnostic technologies. In this study, Plasmodium vivax-specific DNA probes have been identified by carrying out genomic subtractive hybridization. In this approach, the recombinant clones from a P. vivax genomic library are screened with radiolabelled human and P. falciparum DNA. The colonies which react with labelled P. falciparum and human DNA are eliminated and those which do not produce any autoradiographic signal have been subjected to further screening procedures. Three P. vivax specific DNA probes have been obtained by these repeated screenings. Further analyses indicate that these probes are specific and sensitive enough to detect P. vivax infection in clinical blood samples when used in a non-radioactive DNA hybridization assay.
核酸探针在病原微生物检测中的应用已成为诊断技术不可或缺的一部分。在本研究中，通过基因组消减杂交鉴定出了间日疟原虫特异性DNA探针。在这种方法中，用放射性标记的人类DNA和恶性疟原虫DNA筛选间日疟原虫基因组文库中的重组克隆。与标记的恶性疟原虫和人类DNA发生反应的菌落被剔除，而那些未产生任何放射自显影信号的菌落则进行进一步的筛选程序。通过这些重复筛选获得了三种间日疟原虫特异性DNA探针。进一步分析表明，当用于非放射性DNA杂交检测时，这些探针特异性和敏感性足以检测临床血液样本中的间日疟原虫感染。
