中国兽药杂志
中國獸藥雜誌
중국수약잡지
CHINESE JOURNAL OF VETERINARY DRUG
2012年
12期
2-4,8
,共4页
熊媛媛%黄涛%郑佳%漆世华%谢红玲%吴玉石
熊媛媛%黃濤%鄭佳%漆世華%謝紅玲%吳玉石
웅원원%황도%정가%칠세화%사홍령%오옥석
产肠毒素大肠杆菌%纤毛抗原%纯化%抗血清
產腸毒素大腸桿菌%纖毛抗原%純化%抗血清
산장독소대장간균%섬모항원%순화%항혈청
enterotoxigenic Escherichia coli%pilus antigen%purification%antiserum
采用加热法和盐析法对大肠杆菌K88、K99、987P纤毛抗原进行了提取、纯化,并通过SDS—PAGE凝胶电泳对提纯效果进行了检验。将纯化的纤毛抗原免疫家兔,制备了K88、K99、987P纤毛抗原抗血清,用琼脂扩散试验检测了抗血清的效价。结果表明,纯化的K88、K99、987P纤毛为高纯度的纤毛抗原,其分子量分别约为26、18、20kD。K88、K99、987P抗血清的琼扩效价依次为1:32、1:32、1:128,且特异性高。本研究为K88、K99、987P纤毛抗原定量检测及产纤毛大肠杆菌菌株的鉴定奠定了基础。
採用加熱法和鹽析法對大腸桿菌K88、K99、987P纖毛抗原進行瞭提取、純化,併通過SDS—PAGE凝膠電泳對提純效果進行瞭檢驗。將純化的纖毛抗原免疫傢兔,製備瞭K88、K99、987P纖毛抗原抗血清,用瓊脂擴散試驗檢測瞭抗血清的效價。結果錶明,純化的K88、K99、987P纖毛為高純度的纖毛抗原,其分子量分彆約為26、18、20kD。K88、K99、987P抗血清的瓊擴效價依次為1:32、1:32、1:128,且特異性高。本研究為K88、K99、987P纖毛抗原定量檢測及產纖毛大腸桿菌菌株的鑒定奠定瞭基礎。
채용가열법화염석법대대장간균K88、K99、987P섬모항원진행료제취、순화,병통과SDS—PAGE응효전영대제순효과진행료검험。장순화적섬모항원면역가토,제비료K88、K99、987P섬모항원항혈청,용경지확산시험검측료항혈청적효개。결과표명,순화적K88、K99、987P섬모위고순도적섬모항원,기분자량분별약위26、18、20kD。K88、K99、987P항혈청적경확효개의차위1:32、1:32、1:128,차특이성고。본연구위K88、K99、987P섬모항원정량검측급산섬모대장간균균주적감정전정료기출。
To develop a efficient and specific vaccine against the pathogen, the pill of K88, K99, 987P were purified, the antiserum were produced and the titers of pilus antigens were determined quantitatively. K88, K99, 987P pill were extracted and purified by the heating and salting - out. Molecular weights of extracted pill were calculated by SDS -PAGE gel electrophoresis. The specific K88, K99, 987P antiserum were prepared by immunizing rabbits with purified pill. The serum titers were evaluated by an agar diffusion test. Results showed that molecular weights of purified K88, K99 and 987P pill were 26 kD, 18 kD and 20 kD respectively. The titers of K88, K99 and 987P antiserum were 1 : 32, 1 : 32 and 1 : 128 respectively. The results indicated a high specificity of prepared antiserum. This study may provide a solid foundation for quantitative detection of pilus antigens and evidences for identification of Escherichia coli producing pill.