Advanced SearchSearch Tips
Development of Chicken Immunoglobulin Y for Rapid Detection of Cronobacter muytjensii in Infant Formula Powder
facebook(new window)  Pirnt(new window) E-mail(new window) Excel Download
 Title & Authors
Development of Chicken Immunoglobulin Y for Rapid Detection of Cronobacter muytjensii in Infant Formula Powder
Kim, Yesol; Shukla, Shruti; Ahmed, Maruf; Son, Seokmin; Kim, Myunghee; Oh, Sejong;
  PDF(new window)
The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 , having ca. coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.
enzyme-linked immunosorbent assay;egg yolk;Cronobacter muytjensii;IgY;pathogen detection;infant formula powder;
 Cited by
Detection of Cronobacter species in powdered infant formula by probe-magnetic separation PCR, Journal of Dairy Science, 2014, 97, 10, 6067  crossref(new windwow)
Effects of Pure Curry Consumption on Life Span, Body Weight, and Weight of Organs in Mice Transplanted with Cancer Cells, Ecology of Food and Nutrition, 2006, 45, 5, 329  crossref(new windwow)
Akita, E. M. and Nakai, S. (1992) Immunoglobulins from egg yolk: isolation and purification. J. Food Sci. 57, 629- 634. crossref(new window)

Akita, E. M. and Nakai, S. (1993) Comparison of four purification methods for the production of immunoglobulin from eggs laid by hens immunized with an enterotoxigenic E. coli strain. J. Immunol. Methods 160, 207-262. crossref(new window)

Bar-Oz, B., Preminger, A., Peleg, O., Block, C., and Arad, I. (2001) Enterobacter muytjensii infection in the newborn. Acta Paediatr. 90, 356-358. crossref(new window)

Baumgartner, A., Grand, M., Liniger, M., and Iversen, C. (2009) Detection and frequency of Cronobacter spp. (Enterobacter sakazakii) in different categories of ready-to-eat foods other than infant formula. Int. J. Food Microbiol. 136, 189-192. crossref(new window)

Bradford, M. M. (1976) A rapid and sensitive method for quantitation of microgram quantities of protein utilizing the principle of protein-dye-binding. Anal. Biochem. 72, 248-254. crossref(new window)

Calzado, E. G., Garrido, R. M., and Schade, R. (2001) Human haemoclassification by use of specific yolk antibodies obtained after immunization of chickens against human group antigen. Alter. Lab Animal 29, 717-726.

Canadian Council on Animal Care (1993) Guide to the care and use of experimental animals. Downloaded from http:// on 6/1/2011.

Carlander, D., Wilhelmson, M., and Larsson, A. (2003) Immunoglobulin Y levels in egg yolk from three chicken genotypes. Food Agric. Immunol. 15, 35-40. crossref(new window)

Engvall, E. and Perlmann, P. (1971) Enzyme-linked immunosorbent assay (ELISA): quantitative assay of immunoglobulin G. Immunochem. 8, 871-874. crossref(new window)

Friedemann, M. (2007) Enterobacter muytjensii in food and beverages (other than infant formula and milk powder). Int. J. Food Microbiol. 116, 1-10. crossref(new window)

Hansen, P., Scoble, J. A., Hanson, B., and Hoogenraad, N. J. (1998) Isolation and purification of immunoglobulins from chicken eggs using thiophilic interaction chromatography. J. Immunol. Methods 215, 1-7. crossref(new window)

Hochel, I. and Skvor, J. (2009) Characterization of antibodies for the immunochemical detection of E. sakazakii. Czech J. Food Sci. 27, 66-74.

Iverson, C. and Forsythe, S. J. (2003) Risk profiles of Enterobacter muytjensii, an emergent pathogen associated with infant milk formula. Trends Food Sci. Tech. 14, 443-454. crossref(new window)

Kovacs-Nolan, J. and Mine, Y. (2004) Avian egg antibodies: basic and potential applications. Avian Poultry Biol. Rev. 15, 25-46. crossref(new window)

Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685. crossref(new window)

Lai, K. K. (2001) Enterobacter sakazakii infections among neonates, infants, children, and adults: case reports and a review of the literature. Medicine 80, 113-122. crossref(new window)

Lee, A. K., Chang, K. S., Lee, J. Y. Lee, H. J., and Koo, S. N. (2002) Acid stability of anti-Helicobacter pyroli IgY in aqueous polyol solution. J. Biochem. Mol. Biol. 35, 488-493. crossref(new window)

Liu, W., Li, W., Yin, W., Meng, M., Wan, Y., Feng, C., Wang, S., and Xi, R. P. (2010) Preparation of a monoclonal antibody and development of an indirect competitive ELISA for the detection of chlorpromazine residue in chicken and swine liver. J. Sci. Food Agric. 90, 1789-1795.

Munene, C. N. (2004) Development of chicken polyclonal and mouse monoclonal based enzyme immunoassays for the detection of bbb-cyclocitral in catfish pond water. M.S. Thesis, Louisiana State University, pp 59.

Park, S., Shukla, S., Kim, Y., Oh, S., Kim, S. H., and Kim, M. (2012) Development of sandwich enzyme-linked immunosorbent assay for the detection of Cronobacter muytjensii (formerly called Enterobacter sakazakii). Microbiol. Immunol. 56, 472-479. crossref(new window)

Prakash, B. and Suryanarayana, T. (2005) Evaluation of Salmonella gallinarum outer membrane protein based enzyme linked immunosorbent assay for detecting antibodies in vaccinated and infected chicken. Int. J. Poultry Sci. 4, 222-227. crossref(new window)

Schneid, A. S., Rodrigues, K. L., Chemello, D. Tondo, E. C., Ayub, M. A. Z., and Aleixo, J. A. G. (2006) Evaluation of an indirect ELISA for the detection of Salmonella in chicken meat. Brazilian J. Microbiol. 37, 350-355. crossref(new window)

Voller, A. and Bidwell, D. E. (1976) Enzyme immunoassays for antibodies in measles, cytomegalovirus infections and after rubella vaccination. Br. J. Exp. Pathol. 51, 243-247.

Wallmann, J. C., Staak, C., and Luge, E. (1990) Einfache method zur isolierung von immunoglobulin (Y) aus Eiern immunisierter Huhner. J. Vet. Med. B 37, 317-320. crossref(new window)