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目的建立一种特异、敏感、简便的疟疾聚合酶链反应(PCR)快速诊断方法。方法针对疟原虫SSU rRNA基因序列设计3条分别具有属、种特异性的引物,通过多重PCR反应扩增间日疟原虫和恶性疟原虫不同大小的DNA片段。结果 19份恶性疟原虫血样均扩增出长度为400 bp的特定基因片段,16份间日疟原虫扩增出长度为300 bp的特定基因片段,20份健康人群对照血样经PCR扩增均为阴性。结论该多重PCR检测系统具有高效、敏感、特异、稳定、简便等特点,适宜于大批量样品同时检测,对疟疾的快速诊断、鉴别混合感染有较高的应用价值。
Objective To establish a specific, sensitive and simple malaria polymerase chain reaction (PCR) rapid diagnostic method. Methods Three genomic and species specific primers were designed according to the sequence of SSU rRNA gene of Plasmodium, and DNA fragments of Plasmodium vivax and Plasmodium falciparum were amplified by multiplex PCR. Results The specific fragments of 400 bp in length were amplified from 19 samples of Plasmodium falciparum. Specific fragments of 300 bp in length were amplified from 16 samples of Plasmodium vivax and 20 healthy controls were all amplified by PCR negative. Conclusion The multiplex PCR detection system is characterized by high efficiency, sensitivity, specificity, stability and simplicity. It is suitable for the simultaneous detection of large quantities of samples. It has high application value for the rapid diagnosis of malaria and the identification of mixed infections.