Innovative Technology

Innovation

Focusing on the research and development and application of isothermal nucleic acid amplification technology

重组酶介导等温扩增技术检测疟原虫方法的建立和评价_邵雷

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Abstract: Objective To establish a recombinase-aided amplification (RAA) assay for the detection of Plasmodium parasites. Methods Specific primers targeting the conserved 18S small-subunit ribosomal RNA gene (I8SIRNA) of Plasmodium species were designed, and an optimal primer pair was screened and validated to develop a robust RAA system for nucleic acid amplification under isothermal conditions. The established RAA assay was then used to detect standard Plasmodium plasmids as well as four Plasmodium species—Plasmodium falciparum (P. falciparum), Plasmodium vivax (P. vivax), Plasmodium malariae (P. malariae), and Plasmodium ovale (P. ovale)—along with six other blood-transmitted parasites, including Leishmania infantum, Leishmania donovani, Toxoplasma gondii, Entamoeba histolytica, Babesia canis, and Babesia rodhaini. This allowed us to evaluate the sensitivity and specificity of the assay. Results A highly efficient primer pair was identified, enabling complete amplification within 20 minutes at 37°C using this optimized RAA system. The detection limit for the standard plasmid was as low as 10 copies/µL, while the lower limits for P. falciparum, P. vivax, and P. ovale were 10 copies/µL, and for P. malariae, it was 10 copies/µL as well. Importantly, the RAA assay showed no cross-reactivity with the other blood-transmitted parasites, demonstrating excellent specificity. When applied to screen blood samples from foreign student donors, the RAA results matched those obtained by fluorescence quantitative PCR, confirming its reliability. Conclusion We have successfully developed a rapid, simple, and highly specific RAA method for detecting Plasmodium parasites, which will significantly enhance blood screening efforts and provide valuable support for clinical diagnostics in grassroots and remote healthcare settings.

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重组酶介导等温核酸扩增技术检测细棘球绦虫方法的建立及初步应用评价_周鸿让CDC

重组酶介导恒温扩增技术检测疟原虫方法的建立-浙江国旅

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重组酶介导等温核酸扩增技术检测细棘球绦虫方法的建立及初步应用评价_周鸿让CDC

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重组酶介导恒温扩增技术检测疟原虫方法的建立-浙江国旅