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標題: 多殺型巴斯德桿菌脂多醣體抗原合成基因之研究與快速分類之應用
Characterization of The Biosynthesis Genes for Lipopolysaccharide Somatic Antigens of Pasteurella multocida and Application to Rapid-typing
作者: 蔡雨蓁
Tsai, Yu-Chen
關鍵字: Pasteurella multocida;多殺型巴斯德桿菌;lipopolysaccharide;somatic antigen;biosynthesis gene;typing;脂多醣;體抗原;合成基因;分類
出版社: 微生物暨公共衛生學研究所
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Pasteurella multocida (多殺性巴斯德桿菌)為能引起多種重要動物疾病的病原菌,其脂多醣體抗原 (lipopolysaccharide somatic antigen)是主要的毒力因子之一,傳統上可用瓊膠擴散沉澱反應 (AGP)將P. multocida分為16種不同體抗原血清型,但此方法需製備高免血清且操作時間需1-2天,另一方面,目前在文獻中只有少數幾種P. multocida之體抗原之成份與其生合成基因序列被發表。本研究以PCR技術,成功增幅18株P. multocida標準株之體抗原合成基因序列,定序結果發現,依其序列可將P. multocida之體抗原合成基因分為5種 (G1-G5)不同型別,其中G1為能引起家禽霍亂之體抗原1型之禽源P. multocida (A:1);G2為體抗原2型與5型之菌株;G3為體抗原3型及4型之禽源菌株;G4為能引起豬萎縮性鼻炎之體抗原3型菌株及其他體抗原血清型10、11、12、15和16菌株;G5則為分離自人類之體抗原13型菌株。本研究亦利用PCR增幅與限制酶切割圖譜多型性 (restriction fragment length polymorphism,RFLP)來快速區分P. multocida標準株之體抗原型別,此技術並可應用於田間禽源與豬源P. multocida體抗原型別之快速鑑定,本研究為第一篇利用PCR/RFLP來鑑定P. multocida體抗原型別之報導。

Pasteurella multocida is the causative agent of many diseases in a wide range of domestic animals. Its lipopolysaccharide (LPS) somatic antigen is one of key virulence factors. Conventionally, P. multocida can be divided into 16 serotypes based on LPS somatic antigens by using the agar gel diffusion precipitation test (AGP). However, this method needs preparation of serotype-specific antisera, and takes few days to complete the work. To date, only a few of LPS structures and genome sequences for biosynthesis of LPS in P. multocida have been determined. In this study, we used PCR to amplify the biosynthesis genes of lipopolysaccharide somatic antigen of 18 reference strains of P. multocida. Based on sequencing results of the amplified genes, the biosynthesis genes of LPS of P. multocida could be divided into 5 groups (G1-G5). Strains belong to G1 are serotype A:1 which cause fowl cholera. Stains belong to G2 are LPS serotype 2 and 5. Strains belong to G3 are LPS serotype 3 and 4 that cause fowl cholera. Strains belong to G4 are LPS serotype 3 which causes atrophic rhinitis in pigs and other LPS serotypes such as 10, 11, 12, 15, and 16. Strains belong to G5 are LPS serotype 13 that infects humans. This study also develops a PCR/RFLP method for rapid identification of the LPS serotypes of reference strains of P. multocida quickly. Moreover, this method can be used to identify types of somatic antigen of field strains of P. multocida. This is the first report on development of a PCR/PFLP method for rapid identification of the somatic antigen serotypes of P. multocida.
其他識別: U0005-1908201005402200
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