Characterization and Identification of Chicken Eimeria Species Using Matrix-Assisted laser Desorption/Ionization-Time of Flight Mass Spectrometry

• Main Authors: Shu-Ting Kuo, 郭舒婷
• Other Authors: Yi-Yang Lien
• Format: Others
• Language: zh-TW
• Published: 2005
• Online Access: http://ndltd.ncl.edu.tw/handle/33792405665737967581

碩士 === 國立屏東科技大學 === 獸醫學系 === 93 === Coccidiosis, a disease of parasites affected by Eimeria , usually causes the growth-delay or even death of the chicken and the great damages of chicken industry. Several analytical methods of identification about coccidiosis specie such as polymerase chain reaction-linked restrictionfragment length polymorphism ( RFLP ) and Random Amplification of Polymorphic DNA ( RAPD ) have been declared. In this study, we have used proteomic approach to identify coccidiosis species. Three pure strains of oocyst including E. tenella, E. acervulina and E. maxima，and one isolated drug-resistant ( Diclazuril and Maduramcin ) field strain of E. tenella were chosen as the samples. The oocyst are purified by floating method of in saturated sodium chloride solution. The protein were extracted from oocyst by ultrasonic waves or grinding methods, and characterized by MALDI-TOF ( matrix - assisted laser desorption/ionization - time of flight mass spectrometry ). The results indicate there is remarkable difference among the MALDI mass spectra of the three oocyst. However, three protein ions with m/z 3410, 7626, and 8543 are detected in three samples. In the comparison of two different strains of oocyst, the MALDI spectra show great difference in the protein markers of E. tenella and a drug-resistant strain E. tenella. The m/z values of the predominant protein ions are 4808 and 4262 for E. tenella and drug-resistant strain E. tenella, respectively. The results of this research clearly indicate MALDI is not only useful in identifying different species, but also different subtypes. The technique will be used to identify more coccidiosis species in the future.