A Chip-Based Colony Fusion Recombinase Polymerase Amplification Assay for Monitoring of Antimicrobial Resistance Genes and Their Carrying Species in Surface Water

被引:2
作者
Sollweck, Katharina [1 ]
Streich, Philipp [1 ]
Elsner, Martin [1 ]
Seidel, Michael [1 ]
机构
[1] Tech Univ Munich, Inst Hydrochem, Chair Analyt Chem & Water Chem, D-81377 Munich, Germany
来源
ACS ES&T WATER | 2021年 / 1卷 / 03期
关键词
antibiotic resistance; recombinase polymerase amplification; microarray chip; environmental analytics; monitoring; ANTIBIOTIC-RESISTANCE; WASTE-WATER; ESCHERICHIA-COLI; RAPID DETECTION; ISOTHERMAL AMPLIFICATION; QUANTITATIVE PCR; TREATMENT PLANTS; BACTERIA; QUANTIFICATION; SAMPLES;
D O I
10.1021/acsestwater.0c00110
中图分类号
X [环境科学、安全科学];
学科分类号
08 ; 0830 ;
摘要
This work presents a new approach to the rapid characterization of antibiotic resistance in environmental settings. For monitoring purposes and risk assessment, it is important to link the detection of resistance genes to the actual carrying species and their potential pathogenicity for humans. Starting from a bacterial culture plate, this proof of principle study developed a colony-based fusion recombinase polymerase amplification (RPA) reaction that detects species and resistance genes simultaneously within 1.5 h. A first step generates the fusion product by homogeneous RPA, while detection occurs in a second step via heterogeneous asymmetric RPA (haRPA) on a flow-based microarray chip. The assay system successfully discriminated between Escherichia coli colonies carrying blaCTX-M cluster 1 resistance genes and E. coli carrying blaCTX-M genes of other clusters as well as other bacterial species carrying blaCTX-M resistance genes. A threshold value of 17% was determined for the differentiation between positive and negative samples. Analysis of water from the river Lech demonstrated the possibility of addressing real environmental samples. The potential for multiplexing was demonstrated by successful formation of fusion products by homogeneous RPA also in Klebsiella pneumoniae. This study uses RPA for the first time to create molecular fusion products providing a promising tool for future multiplex analyses of antibiotic resistance in the environment.
引用
收藏
页码:584 / 594
页数:11
相关论文
共 65 条
  • [1] Aali R, 2014, INT J PREVENTIVE MED, V5, P887
  • [2] Succinylated Jeffamine ED-2003 coated polycarbonate chips for low-cost analytical microarrays
    Bemetz, Jonas
    Kober, Catharina
    Meyer, Verena K.
    Niessner, Reinhard
    Seidel, Michael
    [J]. ANALYTICAL AND BIOANALYTICAL CHEMISTRY, 2019, 411 (10) : 1943 - 1955
  • [3] Universal direct PCR amplification system: a time-and cost-effective tool for high-throughput applications
    Ben-Amar, Anis
    Oueslati, Souheib
    Mliki, Ahmed
    [J]. 3 BIOTECH, 2017, 7
  • [4] Colony PCR
    Bergkessel, Megan
    Guthrie, Christine
    [J]. LABORATORY METHODS IN ENZYMOLOGY: DNA, 2013, 529 : 299 - 309
  • [5] Statistics review 13: Receiver operating characteristic curves
    Bewick, V
    Cheek, L
    Ball, J
    [J]. CRITICAL CARE, 2004, 8 (06): : 508 - 512
  • [6] Pilot study of seasonal occurrence and distribution of antibiotics and drug resistant bacteria in wastewater treatment plants in Slovakia
    Birosova, Lucia
    Mackulak, Tomas
    Bodik, Igor
    Ryba, Jozef
    Skubak, Jaroslav
    Grabic, Roman
    [J]. SCIENCE OF THE TOTAL ENVIRONMENT, 2014, 490 : 440 - 444
  • [7] Attributable deaths and disability-adjusted life-years caused by infections with antibiotic-resistant bacteria in the EU and the European Economic Area in 2015: a population-level modelling analysis
    Cassini, Alessandro
    Hogberg, Liselotte Diaz
    Plachouras, Diamantis
    Quattrocchi, Annalisa
    Hoxha, Ana
    Simonsen, Gunnar Skov
    Colomb-Cotinat, Melanie
    Kretzschmar, Mirjam E.
    Devleesschauwer, Brecht
    Cecchini, Michele
    Ouakrim, Driss Ait
    Oliveira, Tiago Cravo
    Struelens, Marc J.
    Suetens, Carl
    Monnet, Dominique L.
    Strauss, Reinhild
    Mertens, Karl
    Struyf, Thomas
    Catry, Boudewijn
    Latour, Katrien
    Ivanov, Ivan Nikolaev
    Dobreva, Elina Georgieva
    Tambic Andrasevic, Arjana
    Soprek, Silvija
    Budimir, Ana
    Paphitou, Niki
    Zemlickova, Helena
    Olsen, Stefan Schytte
    Sonksen, Ute Wolff
    Martin, Pille
    Ivanova, Marina
    Lyytikainen, Outi
    Jalava, Jari
    Coignard, Bruno
    Eckmanns, Tim
    Abu Sin, Muna
    Haller, Sebastian
    Daikos, George L.
    Gikas, Achilleas
    Tsiodras, Sotirios
    Kontopidou, Flora
    Toth, Akos
    Hajdu, Agnes
    Guolaugsson, Olafur
    Kristinsson, Karl G.
    Murchan, Stephen
    Burns, Karen
    Dsstat, Patrizio Pezzotti
    Gagliotti, Carlo
    Dumpis, Uga
    [J]. LANCET INFECTIOUS DISEASES, 2019, 19 (01) : 56 - 66
  • [8] European Commission, A European One Health Action Plan against Antimicrobial Resistance (AMR)
  • [9] Exner M., 2015, RISKWA STATUSPAPIER, P19
  • [10] Feilong Sun, 2011, 2011 International Symposium on Water Resource and Environmental Protection (ISWREP), P1983, DOI 10.1109/ISWREP.2011.5893647