2022, Volume 19, Issue 12
Engineering >> 2022, Volume 19, Issue 12 doi: 10.1016/j.eng.2021.03.024
Quantum Dot Nanobeads-Labelled Lateral Flow Immunoassay Strip for Rapid and Sensitive Detection of Salmonella Typhimurium Based on Strand Displacement Loop-Mediated Isothermal Amplification
a Guangdong Provincial Key Laboratory of Microbial Safety and Health & State Key Laboratory of Applied Microbiology Southern China & Institute of Microbiology, Guangdong Academy of Sciences, Guangzhou 510070, China
b State Key Laboratory of Food Science and Technology & National Engineering Research Center for Functional Foods & Synergetic Innovation Center of Food Safety & Joint International Research Laboratory on Food Safety, School of Food Science and Technology, Jiangnan University, Wuxi 214122, China
# These authors contributed equally to this work.
Next Previous
Abstract
Rapid, sensitive, point-of-care detection of pathogenic bacteria is important for food safety. In this study, we developed a novel quantum dot nanobeads-labelled lateral flow immunoassay strip (QBs-labelled LFIAS) combined with strand displacement loop-mediated isothermal amplification (SD-LAMP) for quantitative Salmonella Typhimurium (ST) detection. Quantum dot nanobeads (QBs) served as fluorescence reporters, providing good detection efficiency. The customizable strand displacement (SD) probe was used in LAMP to improve the specificity of the method and prevent by-product capture. Detection was based on a sandwich immunoassay. A fluorescence strip reader measured the fluorescence intensity (FI) of the test (T) line and control (C) line. The linear detection range of the strip was 102–108 CFU·mL−1. The visual limit of detection was 103 CFU·mL−1, indicating that the system was 10-fold more sensitive than AuNPs-labelled test strips. ST specificity was analyzed in accordance with agarose gel outputs of PCR and SD-LAMP. We detected ST in foods with an acceptable recovery of 85%–110%. The method is rapid, simple, almost equipment-free, and suitable for bacterial detection in foods and for clinical diagnosis.
Keywords
Salmonella ; Typhimurium ; Quantum dot nanobeads ; Lateral flow immunoassay strip ; Loop-mediated isothermal amplification ; Strand displacement probe
Figures
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
References
[ 1 ] Deng H, Gao Z. Bioanalytical applications of isothermal nucleic acid amplification techniques. Anal Chim Acta 2015;853:30–45. link1
[ 2 ] Craw P, Balachandran W. Isothermal nucleic acid amplification technologies for point-of-care diagnostics: a critical review. Lab Chip 2012;12(14):2469–86. link1
[ 3 ] Li B, Chen X, Ellington AD. Adapting enzyme-free DNA circuits to the detection of loop-mediated isothermal amplification reactions. Anal Chem 2012;84 (19):8371–7. link1
[ 4 ] Becherer L, Bakheit M, Frischmann S, Stinco S, Borst N, Zengerle R, et al. Simplified real-time multiplex detection of loop-mediated isothermal amplification using novel mediator displacement probes with universal reporters. Anal Chem 2018;90(7):4741–8. link1
[ 5 ] Rodriguez NM, Linnes JC, Fan A, Ellenson CK, Pollock NR, Klapperich CM. Paperbased RNA extraction, in situ isothermal amplification, and lateral flow detection for low-cost, rapid diagnosis of influenza A (H1N1) from clinical specimens. Anal Chem 2015;87(15):7872–9. link1
[ 6 ] Mori Y, Kitao M, Tomita N, Notomi T. Real-time turbidimetry of LAMP reaction for quantifying template DNA. J Biochem Biophys Methods 2004;59 (2):145–57. link1
[ 7 ] Chen Z, Zhang K, Yin H, Li Qi, Wang L, Liu Z. Detection of Salmonella and several common Salmonella serotypes in food by loop-mediated isothermal amplification method. Food Sci Hum Wellness 2015;4(2):75–9. link1
[ 8 ] Dixit KK, Verma S, Singh OP, Singh D, Singh AP, Gupta R, et al. Validation of SYBR green I based closed tube loop mediated isothermal amplification (LAMP) assay and simplified direct-blood-lysis (DBL)-LAMP assay for diagnosis of visceral leishmaniasis (VL). PLoS Negl Trop Dis 2018;12(11):e0006922. link1
[ 9 ] Tangkanchanapas P, Höfte M, De Jonghe K. Reverse transcription loopmediated isothermal amplification (RT-LAMP) designed for fast and sensitive on-site detection of Pepper chat fruit viroid (PCFVd). J Virol Methods 2018;259:81–91. link1
[10] Tomita N, Mori Y, Kanda H, Notomi T. Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products. Nat Protoc 2008;3(5):877–82. link1
[11] Tanner NA, Zhang Y, Evans TC. Visual detection of isothermal nucleic acid amplification using pH-sensitive dyes. Biotechniques 2014;58(2):59–68. link1
[12] Hsieh K, Mage PL, Csordas AT, Eisenstein M, Tom SH. Simultaneous elimination of carryover contamination and detection of DNA with uracil-DNAglycosylase-supplemented loop-mediated isothermal amplification (UDGLAMP). Chem Commun 2014;50(28):3747–9. link1
[13] Kubota R, Alvarez A, Su WW, Jenkins D. FRET-based assimilating probe for sequence-specific real-time monitoring of loop-mediated isothermal amplification (LAMP). Biol Eng Trans 2011;4(2):81–100. link1
[14] Liu W, Huang S, Liu N, Dong D, Yang Z, Tang Y, et al. Establishment of an accurate and fast detection method using molecular beacons in loop-mediated isothermal amplification assay. Sci Rep 2017;7(1):40125. link1
[15] Ball CS, Light YK, Koh CY, Wheeler SS, Coffey LL, Meagher RJ. Quenching of unincorporated amplification signal reporters in reverse-transcription loopmediated isothermal amplification enabling bright, single-step, closed-tube, and multiplexed detection of RNA viruses. Anal Chem 2016;88(7):3562–8. link1
[16] Tanner NA, Zhang Y, Evans TC. Simultaneous multiple target detection in realtime loop-mediated isothermal amplification. Biotechniques 2012;53(2):81–9. link1
[17] Mashooq M, Kumar D, Niranjan AK, Agarwal RK, Rathore R. Development and evaluation of probe based real time loop mediated isothermal amplification for Salmonella: a new tool for DNA quantification. J Microbiol Methods 2016;126:24–9. link1
[18] Nurul Najian AB, Engku Nur Syafirah EAR, Ismail N, Mohamed M, Yean CY. Development of multiplex loop mediated isothermal amplification (m-LAMP) label-based gold nanoparticles lateral flow dipstick biosensor for detection of pathogenic Leptospira. Anal Chim Acta 2016;903:142–8. link1
[19] Wang Y, Li H, Wang Y, Zhang L, Xu J, Ye C. Loop-mediated isothermal amplification label-based gold nanoparticles lateral flow biosensor for detection of Enterococcus faecalis and Staphylococcus aureus. Front Microbiol 2017;8:192. link1
[20] Yang Z, Xu G, Reboud J, Kasprzyk-Hordern B, Cooper JM. Monitoring genetic population biomarkers for wastewater-based epidemiology. Anal Chem 2017;89(18):9941–5. link1
[21] Zhang Li, Chen Y, Cheng N, Xu Y, Huang K, Luo Y, et al. Ultrasensitive detection of viable Enterobacter sakazakii by a continual cascade nanozyme biosensor. Anal Chem 2017;89(19):10194–200. link1
[22] Phillips EA, Moehling TJ, Bhadra S, Ellington AD, Linnes JC. Strand displacement probes combined with isothermal nucleic acid amplification for instrumentfree detection from complex samples. Anal Chem 2018;90(11):6580–6. link1
[23] Ignatov KB, Barsova EV, Fradkov AF, Blagodatskikh KA, Kramarova TV, Kramarov VM. A strong strand displacement activity of thermostable DNA polymerase markedly improves the results of DNA amplification. Biotechniques 2014;57(2):81–7. link1
[24] Li J, Lv Y, Li N, Wu R, Xing M, Shen H, et al. Robust synthesis of bright multiple quantum dot-embedded nanobeads and its application to quantitative immunoassay. Chem Eng J 2019;361:499–507. link1
[25] Li X, Li W, Yang Q, Gong X, Guo W, Dong C, et al. Rapid and quantitative detection of prostate specific antigen with a quantum dot nanobeads-based immunochromatography test strip. ACS Appl Mater Interfaces 2014;6 (9):6406–14. link1
[26] Gunasekera TS, Attfield PV, Veal DA. A flow cytometry method for rapid detection and enumeration of total bacteria in milk. Appl Environ Microbiol 2000;66(3):1228–32. link1
[27] Subramanian A, Irudayaraj J, Ryan T. Mono and dithiol surfaces on surface plasmon resonance biosensors for detection of Staphylococcus aureus. Sens Actuators B Chem 2006;114(1):192–8. link1
[28] Wachiralurpan S, Sriyapai T, Areekit S, Sriyapai P, Thongphueak D, Santiwatanakul S, et al. A one-step rapid screening test of Listeria monocytogenes in food samples using a real-time loop-mediated isothermal amplification turbidity assay. Anal Methods 2017;9(45):6403–10. link1
[29] Ledlod S, Bunroddith K, Areekit S, Santiwatanakul S, Chansiri K. Development of a duplex lateral flow dipstick test for the detection and differentiation of Listeria spp. and Listeria monocytogenes in meat products based on loopmediated isothermal amplification. J Chromatogr B Analyt Technol Biomed Life Sci 2020;1139:121834. link1
[30] Mei X, Zhai X, Lei C, Ye X, Kang Z, Wu X, et al. Development and application of a visual loop-mediated isothermal amplification combined with lateral flow dipstick (LAMP–LFD) method for rapid detection of Salmonella strains in food samples. Food Control 2019;104:9–19. link1
[31] Li J, Zhai L, Bie X, Lu Z, Kong X, Yu Q, et al. A novel visual loop-mediated isothermal amplification assay targeting gene62181533 for the detection of Salmonella spp. in foods. Food Control 2016;60:230–6. link1
[32] Li X, Zhang S, Zhang H, Zhang L, Tao H, Yu J, et al. A loop-mediated isothermal amplification method targets the phoP gene for the detection of Salmonella in food samples. Int J Food Microbiol 2009;133(3):252–8. link1
[33] Srisawat M, Panbangred W. Efficient and specific detection of Salmonella in food samples using a stn-based loop-mediated isothermal amplification method. BioMed Res Int 2015;2015:356401. link1
[34] Fakhr MK, McEvoy JM, Sherwood JS, Logue CM. Adding a selective enrichment step to the iQ-CheckTM real-time PCR improves the detection of Salmonella in naturally contaminated retail turkey meat products. Lett Appl Microbiol 2006;43(1):78–83. link1
[35] Lee H, Hwang J, Park Y, Kwon D, Lee S, Kang I, et al. Immunomagnetic separation and size-based detection of Escherichia coli O157 at the meniscus of a membrane strip. RSC Adv 2018;8(46):26266–70. link1
[36] Telli AE, Dog˘ruer Y. Discrimination of viable and dead Vibrio parahaemolyticus subjected to low temperatures using propidium monoazide-quantitative loop mediated isothermal amplification (PMA-qLAMP) and PMA-qPCR. Microb Pathog 2019;132:109–16. link1
京公网安备 11010502051620号
