Development of an Oligonucleotide Array for Identification of Bacterial Pathogens of Bovine Mastitis

• Main Authors: Chun-Yu Chu, 朱俊育
• Other Authors: Chi-Ching Yang
• Format: Others
• Language: zh-TW
• Published: 2010
• Online Access: http://ndltd.ncl.edu.tw/handle/57444483394659710774

碩士 === 國立屏東科技大學 === 食品科學系所 === 98 === Cow mastitis is caused by a wide variety of microbial pathogens. In this study, an oligonucleotide array was developed to identify the pathogens causing mastitis. These pathogens include Corynebacterium bovis, Enterococcus faecalis, Escherichia coli, Listeria monocytogenes, Mycoplasma bovis, Pseudomonas aeruginosa, Staphylococcus aureus, Staphylococcus chromogenes, Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus hyicus, Staphylococcus intermedius, Staphylococcus simulans, Staphylococcus xylosus, Streptococcus agalactiae, Streptococcus dysgalactiae, Streptococcus parauberis, and Streptococcus uberis. Specific oligonucleotide probes were designed from the 16S-23S rRNA gene intergenic spacer (ITS) region and immobilized on Nylon membrane. The identification method consisted of PCR amplification of the ITS region using bacterial universal primers, followed by hybridization of the digoxigenin-labeled PCR product with oligonucleotide probes on the array. A total of 35 species-specific oligonucleotide probes were immobilized on the array (0.6 cm × 0.5 cm) to identify 18 species of bacteria. After testing a collection of 322 target strains and 154 nontarget strains, the sensitivity and specificity of the array was 100 % and 96.2 %, respectively. The current array has a great potential to rapidly and correctly identify bacteria causing bovine mastitis.