鸭沙门氏菌鞭毛抗原的提取与抗原性鉴定初报

Separation and Primary Antigenicity Identification of Flagellin from Salmonella Anatum

【研究目的】提取沙门氏菌鞭毛蛋白并鉴定其抗原性;【方法】选用鸭沙门氏菌经半固体培养基诱导鞭毛,经肉汤培养后,用酸裂解、经差速离心、硫酸铵沉淀,分离出粗制鞭毛蛋白。将粗制的鞭毛蛋白用SDS-PAGE和磷钨酸负染投射电镜观察,并包板做ELISA检测其抗原性;【结果】SDS-PAGE显示提取的鞭毛蛋白相对分子质量约为58.0KD,且纯度较高,抗原性较好,每1000ml培养液可获得32.8mg鞭毛蛋白;【结论】酸裂解法可获得高质量的鞭毛蛋白,抗原性较好,为禽副伤寒沙门氏菌病诊断方法的建立奠定了基础。

[Objective]To separate and identify the flagellin from Salmonella anatum. [Method]Salmonella anatum was induced with semisolid medium for flagellin, and cultured with M broth, then obtained by centrifuging. The flagellin was separated by acid-splitting and differential centrifugation, ammonium sulate precipitationi. The obtained flagellin was evaluated by sodiummdodecyl sulfate polyacrylamide gel elec- trophoresis （SDS-PAGE）, phosphotungstic acid negative staining and electron microscopy. Antigencity of flagellin was detected by direct ELISA. [Results]SDS-PAGE showed that the purified flagellin protein molecular weight was 58KD; the protein possessed good antigencity and 1000 milliliters of culture pro- duced 32.8mg flagellin. [Conclusion]The flagellin with good antigencity from Salmonella anatum can be obtained by acid-splitting. The purified flagellin may lay the foundation for developping methods for avian paratyphoid diagnosis.