An ELISA-on-a-chip biosensor system coupled with immunomagnetic separation for the detection of Vibrio parahaemolyticus within a single working day.
In this study, we constructed a rapid detection system for a foodborne pathogen, Vibrio parahaemolyticus, by using enzyme-linked immunosorbent assay (ELISA)-on-a-chip (EOC) biosensor technology to minimize the risk of infection by the microorganism. The EOC results showed a detection capability of approximately 6.2x10(5) cells per ml, which was significantly higher than that of the conventional rapid test kit. However, this high level of sensitivity required cultivation of the pathogen prior to analysis, which typically exceeded a day. To shorten the test period, we combined the EOC technology with immunomagnetic separation (IMS), which could enhance the sensitivity of the biosensor. IMS was carried out with magnetic particles coated with a monoclonal antibody specific to the microbe. To test the performance of the IMS-EOC method, fish intestine samples were prepared by artificially inoculating less than 1 or 5 CFU/10 g, allowing for enrichment over predetermined times, and analyzing the sample by using the EOC sensor after concentrating the culture 86-fold via IMS. Using this approach, the bacterium was detected after (at most) 9 h, which approximately corresponds to standard working hours. Thus, the IMS-EOC method allowed for the rapid detection of V. parahaemolyticus, which is responsible for foodborne diseases, and this method could be used for early isolation of contaminated foods before distribution.
Seo
Sung-Min
SM
Program for Bio-Microsystem Technology, Korea University, 1, 5-ka, Anam-dong, Seongbuk-Gu, Seoul 136-701, Republic of Korea.
Cho
Il-Hoon
IH
Jeon
Jin-Woo
JW
Cho
Hyun-Kyu
HK
Oh
Eun-Gyoung
EG
Yu
Hong-Sik
HS
Shin
Soon-Bum
SB
Lee
Hee-Jung
HJ
Paek
Se-Hwan
SH
D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D002138:Calibration; D015169:Colony Count, Microbial; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D006801:Humans; D018189:Immunomagnetic Separation / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014736:Vibrio parahaemolyticus / Q000276:immunology / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal