A versatile microfluidic platform for malaria infection screening and Plasmodium species genotypingResearch in context
Leshan Xiu,
Huimin Li,
Qinqin Hu,
Yuqian Zhang,
Shen-Bo Chen,
Chenxi Wang,
Xiao-Nong Zhou,
Jun-Hu Chen,
Kun Yin
Affiliations
Leshan Xiu
School of Global Health, Chinese Center for Tropical Diseases Research, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China; One Health Center, Shanghai Jiao Tong University-The University of Edinburgh, Shanghai, 200025, China; Corresponding author. 227 South Chongqing Road, Shanghai, 200025, China.
Huimin Li
School of Global Health, Chinese Center for Tropical Diseases Research, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China; One Health Center, Shanghai Jiao Tong University-The University of Edinburgh, Shanghai, 200025, China
Qinqin Hu
School of Global Health, Chinese Center for Tropical Diseases Research, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China; One Health Center, Shanghai Jiao Tong University-The University of Edinburgh, Shanghai, 200025, China
Yuqian Zhang
Department of Surgery, Division of Surgery Research, Mayo Clinic, Rochester, MN, 55905, USA; Microbiome Program, Center for Individualized Medicine, Mayo Clinic, Rochester, MN, 55905, USA
Shen-Bo Chen
National Institute of Parasitic Diseases, Chinese Center for Diseases Control and Prevention (Chinese Center for Tropical Diseases Research), National Health Commission of the People's Republic of China (NHC) Key Laboratory of Parasite and Vector Biology, World Health Organization (WHO) Collaborating Center for Tropical Diseases, National Center for International Research on Tropical Diseases, Shanghai, 200025, China
Chenxi Wang
School of Global Health, Chinese Center for Tropical Diseases Research, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China; One Health Center, Shanghai Jiao Tong University-The University of Edinburgh, Shanghai, 200025, China
Xiao-Nong Zhou
National Institute of Parasitic Diseases, Chinese Center for Diseases Control and Prevention (Chinese Center for Tropical Diseases Research), National Health Commission of the People's Republic of China (NHC) Key Laboratory of Parasite and Vector Biology, World Health Organization (WHO) Collaborating Center for Tropical Diseases, National Center for International Research on Tropical Diseases, Shanghai, 200025, China; Hainan Tropical Diseases Research Center (Hainan Sub-Center, Chinese Center for Tropical Diseases Research), Haikou, 571199, China
Jun-Hu Chen
National Institute of Parasitic Diseases, Chinese Center for Diseases Control and Prevention (Chinese Center for Tropical Diseases Research), National Health Commission of the People's Republic of China (NHC) Key Laboratory of Parasite and Vector Biology, World Health Organization (WHO) Collaborating Center for Tropical Diseases, National Center for International Research on Tropical Diseases, Shanghai, 200025, China; Hainan Tropical Diseases Research Center (Hainan Sub-Center, Chinese Center for Tropical Diseases Research), Haikou, 571199, China; Corresponding author. 207 Rui Jin Er Road, Shanghai, 200025, China.
Kun Yin
School of Global Health, Chinese Center for Tropical Diseases Research, Shanghai Jiao Tong University School of Medicine, Shanghai, 200025, China; One Health Center, Shanghai Jiao Tong University-The University of Edinburgh, Shanghai, 200025, China; Corresponding author. 227 South Chongqing Road, Shanghai, 200025, China.
Summary: Background: Malaria, a widespread parasitic disease caused by Plasmodium species, remains a significant global health concern. Rapid and accurate detection, as well as species genotyping, are critical for effective malaria control. Methods: We have developed a Flexible, Robust, Equipment-free Microfluidic (FREM) platform, which integrates recombinase polymerase amplification (RPA) and clustered regularly interspaced short palindromic repeats (CRISPR)-based detection, enabling simultaneous malaria infection screening and Plasmodium species genotyping. The microfluidic chip enabled the parallel detection of multiple Plasmodium species, each amplified by universal RPA primers and genotyped by specific crRNAs. The inclusion of a sucrose solution effectively created spatial separation between the RPA and CRISPR assays within a one-pot system, effectively resolving compatibility issues. Findings: Clinical assessment of DNA extracts from patients with suspected malaria demonstrates the FREM platform's superior sensitivity (98.41%) and specificity (92.86%), yielding consistent results with PCR-sequencing for malaria detection, which achieved a positive predictive agreement of 98.41% and a negative predictive agreement of 92.86%. Additionally, the accuracy of species genotyping was validated through concordance rates of 90.91% between the FREM platform and PCR-sequencing. Interpretation: The FREM platform offers a promising solution for point-of-care malaria screening and Plasmodium species genotyping. It highlights the possibility of improving malaria control efforts and expanding its applicability to address other infectious diseases. Funding: This work was financially supported by International Joint Laboratory on Tropical Diseases Control in Greater Mekong Subregion, National Natural Science Foundation of China, the Natural Science Foundation of Shanghai, Bill & Melinda Gates Foundation and National Research and Development Plan of China.
