分型基因芯片检测疟原虫的研制及应用

Development of diagnostic gene chip in detection of plasmodium

目的 研制快速检测间日疟原虫和恶性疟原虫等并可进行分型的基因芯片。方法 筛选疟原虫高度保守而且具有种属特异性的SSrRNA基因片断为探针 ,制备疟原虫诊断和分型的基因芯片 ;检测时提取标本中的疟原虫核酸 ,用不对称PCR技术进行扩增和荧光标记 ,然后与检测芯片杂交 ,进行扫描和分析。结果 建立的疟原虫检测芯片能特异和敏感的对间日疟原虫和恶性疟原虫诊断及分型。结论 成功制备用于疟原虫快速侦检和分型的基因芯片 。

Objective To establish the technique of diagnosis gene ch i p for detection of Plasmodium vivax (P.v.), Plasmodium falciparum (P.f.) and mixed infections with P.v.and P.f. Methods According to the conservative and specific sequen ce of the small subunit ribosomal RNA (SSrRNA) fragment of plasmodium,probes wer e designed and synthesized to be spotted on the microarray.After being extracted from sample,the plasmodium nucleotides were amplified and labeled with fluoresc ence Cy3-dUTP by dissymmetrical PCR,then hybridized with the diagnosis gene chip s.Finally,the gene chips were scanned and analyzed. Results The diagnosis gene chips can detect P.v.,P.f.and mixed infections with an adequate sensitivity and specificity.Conclusion Diagnostic gene chips for plasmodium were esta blished successfully and it will be a valuable tool for malaria diagnosis and pr evention.