摘要
龋齿DNA疫苗工程菌采用的大肠杆菌DH-5α在生产过程中极易污染大肠杆菌噬菌体,所以应对原始菌种、主菌种和工作菌种及大量生产时的发酵液作大肠杆菌噬菌体检测。用大肠杆菌噬菌体VCSM13为标准噬菌体株,对大肠杆菌C3000和DH-5α分别作噬菌斑检测和pfu值计算,验证并确定以VCSM13作为标准噬菌体株,C3000作为检测菌株,对龋齿DNA疫苗原始菌种、主菌种(第一代)、工作菌种(2007001)和其发酵液(200703)分别作噬菌体检测,并建立了检测大肠杆菌噬菌体的直接噬菌斑法。结果显示VCSM13在DH-5α的噬菌斑计数为76,pfu/ml为7.6×1013,C3000的噬菌斑计数为81,pfu/ml为8.1×1013,龋齿DNA疫苗的原始菌种、主菌种、工作菌种和发酵液,噬菌斑计数全部为0。Pfu也为0。阳性对照为74,pfu/ml是7.4×1013,阴性对照为0。通过对阳性对照样本作增殖法试验及挑斑接种验证后,证明此法操作简单,灵敏度高。
Engineering bacteria of PGJAP is E. coli DH-5a,E. coli, it is easy to be polluted by E. coli Phage. The original strains, main strains and working strains of PGJAP should be necessary to detected with E. coli Phage. The direct bacteriophage plaque assay was established using E. coli C3000 identified by the standard strains of E. coli phage as hosts. The results showed that the plaque count of DH-5a is 76,pfu/ml is 7.6 × 10^13 ,the plaque count of C3000 is 81 ,pfu/ml is 8.1 × 10^13, All of the plaque count of original strains, main strains and working strains of PGJAP is 0, pfu/ml is 0. The method is highly sensitibe and is easy to handle.
出处
《微生物学免疫学进展》
2010年第2期36-39,共4页
Progress In Microbiology and Immunology
基金
国家科技支撑计划课题研究专项
