Development of a novel biochip for rapid multiplex detection of seven mastitis-causing pathogens in bovine milk samples

• Published in: Journal of veterinary diagnostic investigation Vol. 20; no. 4; pp. 463 - 471
• Main Authors: Lee, K.H, Lee, J.W, Wang, S.W, Liu, L.Y, Lee, M.F, Chuang, S.T, Shy, Y.M, Chang, C.L, Wu, M.C, Chi, C.H
• Format: Journal Article
• Language: English
• Published: United States 01.07.2008
• Subjects: accuracy; animal pathogenic bacteria; animal pathogens; Animals; Bacteria - classification; Bacteriological Techniques; biochips; bovine mastitis; Cattle; dairy cattle; disease detection; disease diagnosis; DNA, Bacterial - isolation & purification; Mastitis, Bovine - microbiology; microbial genetics; Microchip Analytical Procedures - veterinary; milk; Milk - microbiology; nucleic acid hybridization; pathogen identification; polymerase chain reaction; Sensitivity and Specificity; species differences; Species Specificity
• ISSN: 1040-6387; 1943-4936
• DOI: 10.1177/104063870802000408

To efficiently prevent and treat bovine mastitis and minimize its effect on the dairy industry, a sensitive, rapid, and specific test is required for identifying the mastitis-causing pathogens. In this study, a biochip capable of detecting 7 common species of mastitis-causing pathogens, including Corynebacterium bovis, Mycoplasma bovis, Staphylococcus aureus, and the Streptococcus spp. S. agalactiae, S. bovis, S. dysgalactiae, and S. uberis, within 6 hr was developed. The technique is based on DNA amplification of genes specific to the target pathogens and consists of 4 basic steps: DNA extraction of bacteria, polymerase chain reaction, DNA hybridization, and colorimetric reaction. To examine the accuracy and specificity of this biochip, a preliminary test with 82 random quarter milk samples were analyzed and compared with results from conventional microbiological methods conducted simultaneously. Results from all but 1 sample analyzed by the biochip were in agreement with those analyzed by bacteriology. The biochip could be a feasible tool for rapidly diagnosing mastitis-causing pathogens in milk and providing information for a more effective treatment to cure mastitis.