Development of Chicken Immunoglobulin Y for Rapid Detection of Cronobacter muytjensii in Infant Formula Powder

• 발행기관: 한국축산식품학회
• 저자명: Ye Sol Kim Shruti Shukla Maruf Ahmed Seok Min Son Myung Hee Kim Se Jong Oh
• 간행물 정보: 『Food Science of Animal Resources』Korean Journal for Food Science of Animal Resources 제32권 제6호, 706~712쪽, 전체 7쪽
• 주제분류: 농수해양 > 식품과학
• 파일형태: PDF
• 발행일자: 2012.12.31

영문 초록

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 μg/mL, having ca. 108 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 105 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.

키워드

enzyme-Linked immunosorbent assay egg yolk Cronobacter muytjensii IgY pathogen detection infant formula powder