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標題: 雞傳染性鼻炎菌莢膜生合成基因之分析與重組次單位疫苗之研究
Analyses of The Capsule Biosynthesis Genes and Development of Recombinant Subunit Vaccine of Avibacterium paragallinarum
作者: 陳秉誼
Chen, Ping-Yi
關鍵字: Avibacterium paragallinarum;雞傳染性鼻炎菌;Capsule biosynthesis;Recombinant subunit vaccine;莢膜生合成;重組次單位疫苗
出版社: 獸醫微生物學研究所
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雞傳染性鼻炎菌 (Avibacterium paragallinarum) 為引起雞傳染性可利查 (infectious coryza,IC) 之病原菌,此病又可稱為傳染性鼻炎,屬於急性上呼吸道疾病,目前在世界各地皆已造成重大的經濟損失,因為感染此病之蛋雞會造成其產蛋率下降10-40%,感染肉雞或種雞則會引起食慾不振,造成生長遲緩,增加其淘汰率。雖然目前已有不活化之全菌疫苗被廣泛使用,但因為不同血清型之交叉保護效果差,而且製作成本高,在世界各地仍持續爆發A. paragallinarum之病例,因此需要研發具有更高保護力之有效疫苗。本研究目的為探討A. paragallinarum莢膜生合成基因與研發其重組次單位疫苗 (r-P6、r-△P6及r-PD)。實驗中使用2株來自日本之標準菌株,分別為221 (血清型A型) 與 H18 (血清型C型),以及4株台灣野外分離株 (皆為血清型C型),結果顯示A. paragallinarum存在兩種不同莢膜生合成基因型 (type I及II),而且此菌之莢膜型與其血清型無直接相關性。莢膜型type I和type II基因群分別含有6和5個基因 (acbEDGHCB及ccbEF1F2CB),而其轉譯之蛋白質分別與家禽霍亂巴氏德桿菌 (Pasteurella multocida) 血清型F型及D型的莢膜生合成酵素極為相似。以各株A. paragallinarum經不同酵素處理後之血球凝集試驗,以及萃取其莢膜多醣體進行酵素分解試驗,結果證實莢膜基因型type I及type II之A. paragallinarum莢膜成份,分別為軟骨膠醣 (chondroitin) 及肝素醣 (heparosan)。本研究為第一篇關於A. paragallinarum莢膜生合成基因與其莢膜化學成分之報導,此研究結果對於A. paragallinarum致病機轉之瞭解與其疫苗之研發,將有重要助益。而重組次單位疫苗之動物免疫攻毒試驗結果顯示,本研究中所使用的重組次單位疫苗 (r-P6、r-△P6及r-PD) 皆具有降低雞隻臨床症狀發病指數之效力,其中r-P6可提供雞隻至少67% 之保護力,故r-P6具有發展為傳染性可利查有效疫苗之潛力。

Avibacterium (Haemophilus) paragallinarum is the causative agent of infectious coryza (IC), an economically important infectious disease of chickens. The major economic effect of this disease is reduction in egg production (10% - 40%) in layer flocks and increased culling rate in broiler flocks. Although inactivated whole-cell vaccines of A. paragallinarum are widely available, outbreaks of infectious coryza continue to occur and cause significant economic loss worldwide. The aim of this study was to investigate the capsule biosynthesis genes of A. paragallinarum and development of recombinant subunit vaccines (r-P6, r-△P6 and r-PD) for control of infectious coryza. Two reference strains, 221 (serovar A) and H18 (serovar C), together with four Taiwanese field strains (all serovar C), were investigated. The result showed that two genotypes (I and II) of capsule biosynthetic loci were present in A. paragallinarum and the capsule genotypes are independent on the serotypes. The genotypes I and II of capsule biosynthetic loci comprised six and five genes (acbEDGHCB and ccbEF1F2CB), respectively, encoding proteins that were most similar to those found in capsule biosynthetic loci of Pasteurella multocida type F and type D, respectively. This result suggested that the genotypes I and II of A. paragallinarum contain chondroitin and heparosan, respectively. This notion was supported by results derived from enzymatic digestion of capsule materials extracted from A. paragallinarum and by results derived from hemagglutination (HA) tests using bacterial cells treated with different enzymes. To our knowledge, this is the first report showing that different genotypes of capsule biosynthesis genes are present in A. paragallinarum and that chondroitin and heparosan are the chemical component of the capsule of A. paragallinarum. The results of vaccination and challenge tests showed that all recombinant subunit vaccines used in this study can reduce clinical sign scores of chickens. Using r-P6 protein as subunit vaccine can protect at least 67% of chickens against the challenge of the field strain TW07. Therefore, r-P6 is a promising candidate antigen for developing vaccines against infectious coryza.
其他識別: U0005-2207200915332000
Appears in Collections:微生物暨公共衛生學研究所

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