一步法实时荧光RT-PCR在疟原虫检测中的应用  

Application of One-Step qRT-PCR in the Detection of Plasmodium

作　　者：何丽润 曾华颖 郑夔 戴俊 师永霞 孙芳芳 杨可航 蔡扬尧 HE Li-Run;ZENG Hua-Ying;ZHENG Kui;DAI Jun;SHI Yong-Xia;SUN Fang-Fang;YANG Ke-Hang;CAI Yang-Yao(Guangzhou Customs District Techonology Center,Guangzhou 510000)

机构地区：[1]广州海关技术中心,广州510000

出　　处：《中国口岸科学技术》2023年第3期20-24,共5页China Port Science and Technology

摘　　要：本研究探讨了用一步法实时荧光RT-PCR检测疟原虫核酸的可行性。分别用一步法实时荧光RT-PCR和实时荧光PCR两种扩增方法对不同稀释度的样本总RNA和基因组DNA进行检测,同时对20份疟原虫阳性血样进行检测。结果显示,一步法实时荧光RT-PCR可检测到的稀释度最高,比实时荧光PCR高1000倍。运用一步法实时荧光RT-PCR方法检测出20份全部阳性样本,检出率为100%;实时荧光PCR方法检测出18份阳性样本,检出率为90%。结果表明,一步法实时荧光RT-PCR方法用于疟原虫检测比实时荧光PCR方法更具有优势,敏感性更强,准确性更高。This study attempts to explore the feasibility of using one-step RT-PCR to detect nucleic acid of Plasmodium.Two amplification methods,one-step RT-PCR and qPCR,were used to detect the total RNA and genomic DNA of samples with different dilutions,and 20 Plasmodium positive blood samples were detected at the same time.The results showed that the highest dilution could be detected by one-step RT-PCR,which was 1000 times more sensitive than qPCR.A one-step RT-PCR method was used to detect all 20 positive samples,with a detection rate of 100%,and the qPCR method to detect 18 positive samples,with a detection rate of 90%.This indicates that the one-step RT-PCR method for malaria parasite detection has more advantages,stronger sensitivity,and higher accuracy than the qPCR method.

关 键 词：疟原虫 荧光定量PCR法 18S rRNA 

分 类 号：O657.3[理学—分析化学] R446.5[理学—化学]