Your browser doesn't support javascript.
loading
Capture and identification of bacteria from fish muscle based on immunomagnetic beads and MALDI-TOF MS.
Chai, Zhaoliang; Bi, Hongyan.
Afiliação
  • Chai Z; College of Food Science and Engineering, Shanghai Ocean University, Hucheng Ring Road 999, Pudong New District, 201306 Shanghai, China.
  • Bi H; College of Food Science and Engineering, Shanghai Ocean University, Hucheng Ring Road 999, Pudong New District, 201306 Shanghai, China.
Food Chem X ; 13: 100225, 2022 Mar 30.
Article em En | MEDLINE | ID: mdl-35498980
ABSTRACT
In the present study, E. coli was taken as a model bacterium, anti-E. coli functionalized magnetic beads were constructed and used to capture E. coli from aqueous extracts of fish sarcoplasmic protein (FSP) and fish muscle protein of sablefish. The excellency of the reproducibility of the present protocol was demonstrated by capturing E. coli from sablefish FSP extracts. The presence of 10 CFU/mL E. coli is still detectable. A microbial safety test on the surface of fish muscle was successfully performed. The bacterial identification accuracy from samples with different matrices was found to be excellent with RSD = 3%. High specific detection of target bacteria in complex biological samples was testified by spiking Staphylococcus aureus and Klebsiella pneumoniae in samples as interference. Ten biomarker ions were discovered for E. coli's recognition. It is promising to apply the present protocol in bacterial analysis in muscle food samples to ensure their safety.
Palavras-chave
ACN, acetonitrile; Ab, antibody; AuNP@SMBs, gold nanoparticle-coated starch magnetic beads; BSA, bovine serum albumin; CFU, colony forming units; CHCA, α-cyano-4-hydroxycinnamic acid; DI water, deionized water; E. coli, Escherichia coli; EDC, N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide hydro-chloride; EDTA, ethylene diamine tetraacetic acid; Escherichia coli; FSP, fish sarcoplasmic protein; FcMBL@Fe3O4, fragment crystallizable mannose binding lectin-modified Fe3O4; HCl, hydrochloric acid; IgG, immunoglobulin G; IgG@Fe3O4, immunoglobulin G-modified Fe3O4; Janus M-MSNs, Janus magnetic mesoporous silica nanoparticles; LB, Luria-Bertani; MALDI-TOF MS, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry; MBs, magnetic beads; MES, 4-morpholineethanesulfonic acid; MEST, MES with Tween 20; Magnetic beads; OD, optical density; PBS, phosphate buffered saline; PBST, phosphate-buffered saline with Tween 20; PCA, principal component analysis; PCR, polymerase chain reaction; PLS-DA, partial least square-discriminant analysis; RPA, recombinase polymerase amplification; SA, sinapinic acid; SERS, surface-enhanced Raman scattering; SPA, staphylococcus protein; STEC, Shiga toxin-producing Escherichia coli; Sablefish; TFA, trifluoroacetic acid; Tris, tris(hydroxymethyl)aminomethane; anti-E. coli@MBs, anti-Escherichia coli (E. coli) functionalized magnetic beads (MBs) anti-E. coli attached MBs E. coli antibody modified magnetic beads; anti-E. coli@MBs@bacteria, anti-E. coli@MBs attached with bacteria conjugates bacteria captured by anti-E. coli@MBs; m/z, mass-to-charge ratio; matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS); sulfo-NHS, sulfo-N-hydroxysuccinimide

Texto completo: 1 Base de dados: MEDLINE Idioma: En Ano de publicação: 2022 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Idioma: En Ano de publicação: 2022 Tipo de documento: Article