Establishment of a Multiplex Detection Method for Common Bacteria in Blood Based on Human Mannan-Binding Lectin Protein-Conjugated Magnetic Bead Enrichment Combined with Recombinase-Aided PCR Technology

作     者：ZHAO Zi Jin CHEN Xiao Ping HUA Shao Wei LI Feng Yu ZHAO Meng XING Chen Hao WANG Jie TIAN Feng Yu ZHANG Rui Qing LYU Xiao Na HAN Zhi Qiang WANG Yu Xin LI Hong Yi SHEN Xin Xin MA Xue Jun TIE Yan Qing ZHAO Zi Jin;CHEN Xiao Ping;HUA Shao Wei;LI Feng Yu;ZHAO Meng;XING Chen Hao;WANG Jie;TIAN Feng Yu;ZHANG Rui Qing;LYU Xiao Na;HAN Zhi Qiang;WANG Yu Xin;LI Hong Yi;SHEN Xin Xin;MA Xue Jun;TIE Yan Qing

作者机构：Graduate SchoolHebei North UniversityZhangjiakou 075000HebeiChina Department of Clinical LaboratoryHebei General HospitalShijiazhuang 050051HebeiChina National Key Laboratory of Intelligent Tracking and Forecasting for Infectious DiseasesNHC Key Laboratory of Medical Virology and Viral DiseasesNational Institute for Viral Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China National Institute for Communicable Disease Control and PreventionChinese Center for Disease Control and PreventionBeijing 102206China Department of Clinical Laboratory Pathology Diagnostic CenterShiyan General HospitalShiyan 442000HubeiChina Graduate School Hebei Medical UniversityShijiazhuang 050031HebeiChina 

出 版 物：《Biomedical and Environmental Sciences》 (生物医学与环境科学（英文版）)

年 卷 期：2024年第37卷第4期

页      面：387-398页

核心收录：

学科分类：0830[工学-环境科学与工程（可授工学、理学、农学学位）] 1004[医学-公共卫生与预防医学(可授医学、理学学位)] 1010[医学-医学技术（可授医学、理学学位）] 10[医学] 

基　　金：funded by the National Key R&D Program of China[2021YFC2301102] National Natural Science Foundation of China Key R&D Program of Hebei Province[223777100D] 

主　　题：Staphylococcus aureus Pseudomonas aeruginosa Acinetobacter baumannii Human Mannan-binding lectin protein Bloodstream infection Recombinase-aided PCR assay Multiple detection 

摘      要：Objective Recombinase-aided polymerase chain reaction(RAP)is a sensitive,single-tube,two-stage nucleic acid amplification *** study aimed to develop an assay that can be used for the early diagnosis of three types of bacteremia caused by Staphylococcus aureus(SA),Pseudomonas aeruginosa(PA),and Acinetobacter baumannii(AB)in the bloodstream based on recombinant human mannanbinding lectin protein(M1 protein)-conjugated magnetic bead(M1 bead)enrichment of pathogens combined with *** Recombinant plasmids were used to evaluate the assay *** blood influenza bacteria were used for the specific *** and clinical plasma samples were enriched with M1 beads and then subjected to multiple recombinase-aided PCR(M-RAP)and quantitative PCR(qPCR)*** analysis was used to evaluate the consistency between the two *** The M-RAP method had sensitivity rates of 1,10,and 1 copies/μL for the detection of SA,PA,and AB plasmids,respectively,without cross-reaction to other bacterial *** M-RAP assay obtained results for10 CFU/mL pathogens in the blood within 4 h,with higher sensitivity than qPCR.M-RAP and qPCR for SA,PA,and AB yielded Kappa values of 0.839,0.815,and 0.856,respectively(P0.05).Conclusion An M-RAP assay for SA,PA,and AB in blood samples utilizing M1 bead enrichment has been developed and can be potentially used for the early detection of bacteremia.