A direct,sensitive and high-throughput genus and species-specific molecular assay for large-scale malaria screening

作     者：Yaling Zhao Ye Zhao Yu Sun Lihua Fan Duoquan Wang Heng Wang Xiaodong Sun Zhi Zheng Yaling Zhao;Ye Zhao;Yu Sun;Lihua Fan;Duoquan Wang;Heng Wang;Xiaodong Sun;Zhi Zheng

作者机构：Institute of Basic Medical SciencesChinese Academy of Medical SciencesSchool of Basic MedicinePeking Union Medical CollegeBeijingChina Yunnan Institute of Parasitic DiseasesYunnan Center for Malaria ResearchInstitute of Vector and Pathogen BiologyDali UniversityPuerYunnanChina National Institute of Parasitic DiseasesChinese Center for Disease Control and PreventionWHO Collaborating Center for MalariaSchistosomiasisand FilariasisKey Laboratory of Parasite and Vector BiologyMinistry of HealthShanghaiChina Blood Transfusion DepartmentPeking Union Medical College HospitalChinese Academy of Medical SciencesBeijingChina 

出 版 物：《Infectious Diseases of Poverty》 (贫困所致传染病（英文）)

年 卷 期：2022年第11卷第2期

页      面：11-22页

核心收录：

学科分类：1004[医学-公共卫生与预防医学(可授医学、理学学位)] 1002[医学-临床医学] 1001[医学-基础医学(可授医学、理学学位)] 1010[医学-医学技术（可授医学、理学学位）] 100201[医学-内科学(含：心血管病、血液病、呼吸系病、消化系病、内分泌与代谢病、肾病、风湿病、传染病)] 10[医学] 

基　　金：The National S&T Major Program of China Grant(2018ZX10101001) National Natural Science Foundation of China Grant(81271926) the PUMC Scholar fund from the Chinese Academy of Medical Sciences,CAMS Innovation Fund for Medical Sciences(2018-I2M-1-001) a grant from Oversees Expertise Introduction Center for Discipline Innovation("111 Center")(BP 0820029) 

主　　题：Infectious disease Malaria Molecular screening CLIP-PCR Genus Species High-throughput 

摘      要：Background:Infectious disease diagnostics often requires sensitive molecular assays that identify at both genus and species *** large scale screening,such as malaria screening for elimination,diagnostic assay can be a challenge,as both the throughput and cost of the assay must be *** requirement of nucleic acid extraction hampers the throughput of most molecular ***-amplification of multiple species or multiplex identification either can result in missed diagnosis or are too costly for large-scale screening.A genus-and species-specific diagnostic assay with simplified procedure,high sensitivity and throughput is still *** study aimed to develop a sensitive and high-throughput approach for large-scale infectious disease ***:We developed multi-section Capture and Ligation Probe PCR(mCLIP-PCR)for the direct detection of RNA without extraction and reverse *** tailed sandwich hybridization probes were used to bind at genus-and species-specific sections of the target RNA to cooperatively capture the target onto a 96-well *** enzymatic ligation of the bound probes,a single-stranded DNA formed at each section with distinct tail sequence at the *** were separately PCR-amplified with primers corresponding to tail sequences for genus or species *** applied the method to the active screening ofPlasmodium infections of 4,580 asymptomatic dried blood spot samples collected in malaria endemic areas and compared the results with standard qPCR using linear ***:With multi-section cooperative capture but separate amplification strategy,we accurately identified genusPlasmodium and *** *** without RNA extraction,with favorable sensitivities among the published *** the active screening,our method identified all 53 positive infections including two mixed infections,and *** infections that were missed by standard ***:mCLIP-PCR provide