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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

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Biomedical and Environmental Sciences 2024年第4期37卷 387-398页

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 Graduate School Hebei North UniversityZhangjiakou 075000HebeiChina Department of Clinical Laboratory Hebei General HospitalShijiazhuang 050051HebeiChina National Key Laboratory of Intelligent Tracking and Forecasting for Infectious Diseases NHC 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 Prevention Chinese Center for Disease Control and PreventionBeijing 102206China Department of Clinical Laboratory Pathology Diagnostic Center Shiyan General HospitalShiyan 442000HubeiChina Graduate School Hebei Medical University Shijiazhuang 050031HebeiChina

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 fordetection of bacteremia.

关键词： Staphylococcus aureus Pseudomonas aeruginosa Acinetobacter baumannii Human Mannan-binding lectin protein Bloodstream infection Recombinase-aided PCR assay multiple detection

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Disposable paper-based bipolar electrode array for multiplexed electrochemiluminescence detection of pathogenic DNAs

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Science China Chemistry 2015年第5期58卷 810-818页

作者： Qiumei Feng Hongyuan Chen Jingjuan Xu State Key Laboratory of Analytical Chemistry for Life Science School of Chemistry and Chemical Engineering Nanjing University

A novel disposable paper-based bipolar electrode （BPE） array is fabricated for multiplexed electrochemiluminescence （ECL） detection of pathogenic DNAs. This proposed BPE array device consists of 15 units, each consisting of six sensing cells and two reporting cells patterned using hydrophobic wax. A hairpin structure DNA assembled on the cathodes of BPEs hybridizes with Pt nanoparticles （NPs） labeled probe DNA in the presence of complementary target DNA. The introduction of Pt NPs catalyzes the reduction of dissolved 02 at cathodes and induces an enhanced ECL signal from Ru（bpy）32＋/tripropylamine （TPrA） at the anodes of BPEs. The dissolved 02 lost in reduction reaction could be promptly replenished due to the relatively large contact area of the paper-based cells with air, which ensures the stability of ECL signal. This obtained paper-based BPE array sensor showed excellent performances for the multiplexed analysis of the syphilis （Treponema pallidum） gene, the immunodeficiency virus gene （HIV） and hepatitis B virus gene （HBV）.

关键词： paper-based device bipolar electrode array pathogenic DNAs multiple detection electrochemiluminescence

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Fluorescent aptasensor for detection of live foodborne pathogens based on multicolor perovskite-quantum-dot-encoded DNA probes and dual-stirring-bar-assisted signal amplification

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Journal of Pharmaceutical Analysis 2022年第6期12卷 913-922页

作者： Liu Liu Juncheng Hong Wenhai Wang Shu Xiao Hongzhen Xie Qiqin Wang Ning Gan School of Material Science and Chemical Engineering Ningbo UniversityNingboZhejiang315211China Institute of Pharmaceutical Analysis College of PharmacyJinan UniversityGuangzhou510632China

In this study,a fluorescent(FL)aptasensor was developed for on-site detection of live Salmonella typhimurium(S.T.)and Vibrio parahaemolyticus(V.P.).Complementary DNA(cDNA)of aptamer(Apt)-functionalized multicolor polyhedral oligomeric silsesquioxane-perovskite quantum dots(cDNA-POSSPQDs)were used as encoded probes and combined with dual-stirring-bar-assisted signal amplification for pathogen *** this system,bar 1 was labeled with the *** ***,and then bar 2 was functionalized with *** *** *** introduced,pathogen-Apt complexes would form and be released into the supernatant from bar *** agitation,the two complexes reached bar 2 and subsequently reacted with cDNA-POSS-PQDs,which were immobilized on ***,the encoded probes would be detached from bar 2 to generate FL signals in the ***,the pathogens can resume their free state and initiate next *** swim between the two bars,and the FL signals can be gradually enhanced to maximum after several *** FL signals from released encoded probes can be used to detect the *** particular,live pathogens can be distinguished from dead ones by using an *** detection limits and linear range for *** *** 30 and 10 CFU/mL and 10^(2) -10^(6) CFU/mL,***,this assay has broad application potential for simultaneous on-site detection of various live pathogenic bacteria in water.

关键词： Aptasensor Live bacteria multiple detection Multicolor perovskite quantum dots Dual-stirring-bar

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Recent advances in immunoassays and biosensors for mycotoxins detection in feedstuffs and foods

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Journal of Animal Science and Biotechnology 2022年第2期13卷 365-383页

作者： Runxian Li Yang Wen Fenglai Wang Pingli He State Key Laboratory of Animal Nutrition College of Animal Science and TechnologyChina Agricultural UniversityBeijing 100193China

Mycotoxins are secondary metabolites produced by *** mycotoxin species are highly toxic and are frequently found in cereals and ***,powerful detection methods are vital and effective ways to prevent feed *** detection methods can no longer meet the needs of massive,real-time,simple,and fast mycotoxin *** detection methods based on advanced material and sensor technology are the future *** this review,we highlight recent progress of mycotoxin rapid detection strategies in feedstuffs and foods,especially for simultaneous multiplex mycotoxin ***,biosensors,and the prominent roles of nanomaterials are *** principles of different types of recognition and signal transduction are explained,and the merits and pitfalls of these methods are ***,limitations and challenges of existing rapid sensing strategies and perspectives of future research are discussed.

关键词： Biosensors Immunoassays multiple detection Mycotoxins Nanomaterials Rapid detection

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Tb^(3+)-xylenol orange complex-based colorimetric and luminometric dual-readout sensing platform for dipicolinic acid and metal ions

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Chinese Chemical Letters 2023年第1期34卷 555-559页

作者： Lixia Lu Xinyu Shao Xiaoxiao Lin Linhe Ding Bingchan Song Jian Sun Shandong Provincial Key Laboratory of Animal Resistance Biology Institute of Biomedical SciencesKey Laboratory of Food Nutrition and Safety of Shandong Normal UniversityCollege of Life ScienceShandong Normal UniversityJi nan 250014China State Key Laboratory of Electroanalytical Chemistry Changchun Institute of Applied ChemistryChinese Academy of SciencesChangchun 130022China

A dual-readout sensing platform based on two signal transduction channels can integrate the unique advantages of each sensing pattern,compensate for the deficiency in the adaptive capacity,and enable a more convincing performance in analytical ***,we introduce a responsive molecule dye,xylenol orange(XO),to combine with lanthanide terbium ions(Tb^(3+)).The resultant Tb^(3+)-XO complex exhibited tunable optical properties and was used as a novel colorimetric and luminometric dual-readout sensing platform for assaying the anthrax biomarker,dipicolinic acid(DPA).In the presence of Tb^(3+),the XO solution underwent a color change from yellow to magenta;however,upon adding DPA,the color changed back to yellow immediately,accompanied by the characteristic luminescence emission of Tb^(3+).Considering the strong affinity between DPA/XO and metal ions,the proposed sensing platform was further employed for the determination and differentiation of certain metal ions using linear discriminant *** convenient dual-readout sensing platform offers several notable features and significantly promotes the application and development of lanthanide-based materials.

关键词： Lanthanide complex Xylenol orange Bacillus anthracis multiple detection Linear discriminant analysis

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