Abstract
Salmonella is one of the most common pathogenic bacterium, which causes food spoilage and human diseases. The national standard detection method (GB4789.4) and other fast detection methods for Salmonella cannot satisfy the requirements in the present climate because of their inefficiency, low accuracy, high equipment requirements, or high cost. In this study, we combine isothermal recombinase polymerase amplification (RPA) with unmodified gold nanoparticles (AuNPs) to detect Salmonella in milk. A rapid, effective, visualized, and low-cost RPA-AuNP assay was developed, with a detection limit of 50 CFU for milk samples after enrichment for 6 h or 1 pg for DNA within 15 min at 37 °C using simple water bath equipment. Furthermore, it exhibited no cross-reactions with other pathogens by using highly specific region of invA of Salmonella as the target detection gene. And it may apply to the local food inspection and detection center whose equipment conditions are poor.
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References
Alzwghaibi AB, Yahyaraeyat R, Fasaei BN, Langeroudi AG, Salehi TZ (2018) Rapid molecular identification and differentiation of common Salmonella serovars isolated from poultry, domestic animals and foodstuff using multiplex PCR assay. Arch Microbiol 200:1009–1016. https://doi.org/10.1007/s00203-018-1501-7
Bai J, Trinetta V, Shi X, Noll LW, Magossi G, Zheng W, Porter EP, Cernicchiaro N, Renter DG, Nagaraja TG (2018) A multiplex real-time PCR assay, based on invA and pagC genes, for the detection and quantification of Salmonella enterica from cattle lymph nodes. J Microbiol Methods 148:110–116. https://doi.org/10.1016/j.mimet.2018.03.019
Brewster JD, Paul M (2016) Short communication: improved method for centrifugal recovery of bacteria from raw milk applied to sensitive real-time quantitative PCR detection of Salmonella spp. J Dairy Sci 99(5):3375–3379. https://doi.org/10.3168/jds.2015-9655
Chen X, Wu X, Gan M, Xu F, He L, Yang D, Xu H, Shah NP, Wei H (2015) Rapid detection of Staphylococcus aureus in dairy and meat foods by combination of capture with silica-coated magnetic nanoparticles and thermophilic helicase-dependent isothermal amplification. J Dairy Sci 98(3):1563–1570. https://doi.org/10.3168/jds.2014-8828
Chen Y, Cheng N, Xu Y, Huang K, Luo Y, Xu W (2016) Point-of-care and visual detection of P. aeruginosa and its toxin genes by multiple LAMP and lateral flow nucleic acidbiosensor. Biosens Bioelectron 81:317–323. https://doi.org/10.1016/j.bios.2016.03.006
Chiang YC, Wang HH, Ramireddy L, Chen HY, Shih CM, Lin CK, Tsen HY (2018) Designing a biochip following multiplex polymerase chain reaction for the detection of Salmonella serovars Typhimurium, Enteritidis, Infantis, Hadar, and Virchow in poultry products. J Food Drug Anal 26(1):58–66. https://doi.org/10.1016/j.jfda.2016.11.019
Chunglok W, Wuragil DK, Oaew S, Somasundrum M, Surareungchai W (2011) Immunoassay based on carbon nanotubes-enhanced ELISA for Salmonella enterica serovar Typhimurium. Biosens Bioelectron 26(8):3584–3589. https://doi.org/10.1016/j.bios.2011.02.005
Cunha-Neto AD, Carvalho LA, Carvalho RCT, Dos Prazeres Rodrigues D, Mano SB, Figueiredo EES, Conte-Junior CA (2018) Salmonella isolated from chicken carcasses from a slaughterhouse inthe state of Mato Grosso, Brazil: antibiotic resistance profile, serotyping, and characterization by repetitive sequence-based PCR system. Poult Sci 97(4):1373–1381. https://doi.org/10.3382/ps/pex406
Du XJ, Zang YX, Liu HB, Li P, Wang S (2018) Recombinase polymerase amplification combined with lateral flow strip for Listeria monocytogenes detection in food. J Food Sci 83(4):1041–1047. https://doi.org/10.1111/1750-3841.14078
Garrido-Maestu A, Azinheiro S, Carvalho J, Abalde-Cela S, Carbó-Argibay E, Diéguez L, Piotrowski M, Kolen'ko YV, Prado M (2017) Combination of microfluidic loop-mediated isothermal amplification with gold nanoparticles for rapid detection of Salmonella spp. in food samples. Front Microbiol 8:2159. https://doi.org/10.3389/fmicb.2017.02159 eCollection
Gatto F, Bassani N, Rosa SF, Lievens A, Brustio R, Kreysa J, Querci M (2017) Semi-quantification of GM maize using ready-to-use RTi-PCR plates. Food Anal Methods 10:549–558. https://doi.org/10.1007/s12161-016-0609-0
Kim Y, Yi J (2008) In-situ observation of deposition of gold nanoparticles on the amine-functionalized surface by open liquid-AFM[J]. Korean J Chem Eng 25(2):383–385
Kong C, Wang Y, Fodjo EK, Yang GX, Han F, Shen XS (2017) Loop-mediated isothermal amplification for visual detection of Vibrio parahaemolyticus using gold nanoparticles. Mikrochim Acta 185(1):35. https://doi.org/10.1007/s00604-017-2594-25944
Kumar BK, Raghunath P, Devegowda D, Deekshit VK, Venugopal MN, Karunasagar I, Karunasagar I (2011) Development of monoclonal antibody based sandwich ELISA for the rapid detection of pathogenic Vibrio parahaemolyticus in seafood. Int J Food Microbiol 145(1):244–249. https://doi.org/10.1016/j.ijfoodmicro.2010.12.030
Li SK, Chen AY, Chai YQ, Yuan R, Zhuo Y (2016) Electrochemiluminescence aptasensor based on cascading amplification of nicking endonuclease-assisted target recycling and rolling circle amplifications for mucin 1 detection. Electrochim Acta 212:767–774. https://doi.org/10.1016/j.electacta.2016.07.074
Liu Q, Nam J, Kim S, Lim CT, Park MK, Shin Y (2016) Two-stage sample-to-answer system based on nucleic acid amplification approach for detection of malaria parassites. Biosens Bioelectron 82:1–8. https://doi.org/10.1016/j.bios.2016.03.050
Liu S, Geng Y, Liu L, Sun X, Shao J, Han B, Wang J, Tan K (2018) Development of an isothermal amplification-based assay for the rapid detection of Cronobacter spp. J Dairy Sci 101(6):4914–4922. https://doi.org/10.3168/jds.2017-13931
Mi LJ, Zhu HP, Zhang XD, Hu J, Fan CH (2007) Mechanism of the interaction between Au nano-particles and polymerase in nanoparticle PCR[J]. Chin Sci Bull 52(17):2345–2349
Mirhosseini SA, Fooladi AAI, Amani J, Sedighian H (2017) Production of recombinant flagellin to develop ELISA-based detection of Salmonella Enteritidis. Braz J Microbiol 48(4):774–781. https://doi.org/10.1016/j.bjm.2016.04.033
Mondal B, Ramlal S, Lavu PS, N B, Kingston J (2018) Highly sensitive colorimetric biosensor for Staphylococcal enterotoxin B by a label-free aptamer and gold nanoparticles. Front Microbiol 9:179. https://doi.org/10.3389/fmicb.2018.00179 eCollection 2018
Nguyen TH, Nguyen TD, Ly NH, Kwak CH, Huh YS, Joo SW (2018) On-site detection of sub-mg/kg melamine mixed in powdered infant formula and chocolate using sharp-edged gold nanostar substrates. Food Addit Contam Part A Chem Anal Control Expo Risk Assess 1:1–10. https://doi.org/10.1080/19440049.2018.1466399
Ning Y, Gao Q, Zhang X, Wei K, Chen L (2016) A graphene oxide-based sensing platform for the determination of methicillin-resistant Staphylococcus aureus based on strand-displacement polymerization recycling and synchronous fluorescent signal amplification. J Biomol Screen 21(8):851–857. https://doi.org/10.1177/1087057116653564
Park S, Shukla S, Kim Y, Oh S, Hun Kim S, Kim M (2012) Development of sandwich enzyme-linked immunosorbent assay for the detection of Cronobacter muytjensii (formerly called Enterobacter sakazakii). Microbiol Immunol 56(7):472–479. https://doi.org/10.1111/j.1348-0421.2012.00466.x
Paternò A, Verginelli D, Bonini P, Misto M, Quarchioni C, Dainese E, Peddis S, Fusco C, Vinciguerra D, Marchesi U (2018) In-house validation and comparison of two wheat (Triticum aestivum) taxon-specific real-time PCR methods for GMO quantification supported by droplet digital PCR. Food Anal Methods 11:1281–1290. https://doi.org/10.1007/s12161-017-1097-6
Ranjbar R, Naghoni A, Farshad S, Lashini H, Najafi A, Sadeghifard N, Mammina C (2014) Useof TaqMan real-time PCR for rapid detection of Salmonella enterica serovar Typhi. Acta Microbiol Immunol Hung 61(2):121–130. https://doi.org/10.1556/AMicr.61.2014.2.3
Shem PM, Sardar R, Shumaker-Parry JS (2009) One-step synthesis of phosphine-stabilized gold nanoparticles using the mild reducing agent 9-BBN. Langmuir 25(23):13279–13283. https://doi.org/10.1021/la903003n
Tanguay F, Vrbova L, Anderson M, Whitfield Y, Macdonald L, Tschetter L, Hexemer A, Salmonella Reading Investigation Team (2017) Outbreak of Salmonella reading in persons of eastern Mediterranean origin in Canada, 2014-2015. Can Commun Dis Rep 43(1):14–20
Wang M, Yang J, Gai Z, Huo S, Zhu J, Li J, Wang R, Xing S, Shi G, Shi F, Zhang L (2018a) Comparison between digital PCR and real-time PCR in detection of Salmonella typhimurium in milk. Int J Food Microbiol 266:251–256. https://doi.org/10.1016/j.ijfoodmicro.2017.12.011
Wang Y, Yan W, Wang Y, Xu J, Ye C (2018b) Rapid, sensitive and reliable detection of Klebsiella pneumoniae by label-free multiple crossdisplacement amplification coupled with nanoparticles-based biosensor. J Microbiol Methods 149:80–88. https://doi.org/10.1016/j.mimet.2018.05.003
Wilson R (2018) The use of gold nanoparticles in diagnostics and detection[J]. Chem Soc Rev 37(9):2028–2045
Wu Z, Jin R (2010) On the ligand's role in the fluorescence of gold nanoclusters. Nano Lett 10(7):2568–2573. https://doi.org/10.1021/nl101225f
Yoo CI, Seo D, Chung BH, Chung IS, Song H (2009) A facile one-pot synthesis of hydroxyl-functionalized gold polyhedrons by a surface regulating copolymer[J]. Chem Mater 21(5):13–23
Zhang X, Li M, Zhang B, Chen K, He K (2016) Development of a sandwich ELISA for EHEC O157:H7 Intimin γ1. PLoS One 11(9):e0162274. https://doi.org/10.1371/journal.pone.0162274
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Zhi-guang Chen declares that he has no conflict of interest. Hai-xia Zhong declares that she has no conflict of interest. Huan Luo declares that she has no conflict of interest. Ren-yu Zhang declares that she has no conflict of interest. Jun-rong Huang declares that she has no conflict of interest.
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Hai-xia Zhong is a co-first author
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Chen, Zg., Zhong, Hx., Luo, H. et al. Recombinase Polymerase Amplification Combined with Unmodified Gold Nanoparticles for Salmonella Detection in Milk. Food Anal. Methods 12, 190–197 (2019). https://doi.org/10.1007/s12161-018-1351-6
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DOI: https://doi.org/10.1007/s12161-018-1351-6