title: Immunoliposome-based immunomagnetic concentration and separation assay for rapid detection of Cronobacter sakazakii.
abstract: This study aimed to develop an immunoliposome-based immunomagnetic concentration and separation assay for the rapid detection of Cronobacter sakazakii (C. sakazakii), an acute opportunistic foodborne pathogenic bacterium, in both pure culture and infant formula. To develop the assay, magnetic nanoparticles (diameter 30 nm) were coated with immunoglobulin G (IgG), specifically anti-C. sakazakii IgG, and applied for the sensitive and efficient detection of C. sakazakii using immunoliposomes. The binding efficiency of anti-C. sakazakii IgG to the magnetic nanoparticles was 86.23 ± 0.59%. The assay developed in this study detected as few as 3.3 × 10(3) CFUmL(-1) of C. sakazakii in pure culture within 2h 30 min; in comparison, an indirect non-competitive enzyme-linked immunosorbent assay was able to detect 6.2 × 10(5) CFUmL(-1) of C. sakazakii in pure culture after 17 h. The developed assay did not show any cross-reactivity with other Cronobacter spp. or pathogens belonging to other genera. In addition, the method was able to detect 10(3) CFUmL(-1) of C. sakazakii in infant formula without any pre-incubation. These results confirm that the immunoliposome-based immunomagnetic concentration and separation assay may facilitate highly sensitive, efficient, and rapid detection of C. sakazakii.
authors.affiliation: Department of Food Science and Technology, Yeungnam University, 280 Daehak-ro, Gyeongsan-si, Gyeongsangbuk-do 712-749, Republic of Korea.
authors.forename: Shruti
authors.initials: S
authors.lastname: Shukla
authors.affiliation: Department of Food Science and Technology, Yeungnam University, 280 Daehak-ro, Gyeongsan-si, Gyeongsangbuk-do 712-749, Republic of Korea.
authors.forename: Gibaek
authors.initials: G
authors.lastname: Lee
authors.affiliation: Department of Food Science and Technology, Yeungnam University, 280 Daehak-ro, Gyeongsan-si, Gyeongsangbuk-do 712-749, Republic of Korea.
authors.forename: Xinjie
authors.initials: X
authors.lastname: Song
authors.affiliation: Food Standard Research Center, Korea Food Research Institute, Anyangpangyo-ro 62, 1201 Beon-gil, Seongnam-si, Gyeonggi-do 13539, Republic of Korea.
authors.forename: Sunhyun
authors.initials: S
authors.lastname: Park
authors.affiliation: Department of Food Science and Technology, Yeungnam University, 280 Daehak-ro, Gyeongsan-si, Gyeongsangbuk-do 712-749, Republic of Korea. Electronic address: foodtech@ynu.ac.kr.
authors.forename: Myunghee
authors.initials: M
authors.lastname: Kim
mesh_terms: D000818:Animals; D058491:Bacterial Load / Q000295:instrumentation*; D003199:Computer Systems; D044083:Cronobacter sakazakii / Q000276:immunology* / Q000302:isolation & purification*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D007074:Immunoglobulin G / Q000737:chemistry / Q000276:immunology*; D018189:Immunomagnetic Separation / Q000295:instrumentation*; D008081:Liposomes / Q000276:immunology*; D058185:Magnetite Nanoparticles / Q000737:chemistry; D011817:Rabbits; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D007074:Immunoglobulin G; D008081:Liposomes; D058185:Magnetite Nanoparticles
keywords: Anti-C. sakazakii IgG; C. sakazakii; Immunoliposome; Iron oxide magnetic particles