摘要
目的制备基因转染(D3)细胞并测定能引起先天性感染的几种病原体的敏感性。方法HEP-2细胞的DNA转染人胚肺细胞(HEL),得到基因转染细胞,命名为D3细胞。分析D3细胞的核型。研究丹参对D3细胞促增殖作用和丹参对病原体不同的促增殖作用。普通显微镜观察病原体感染D3细胞后的细胞病变。免疫荧光试验研究D3细胞对病原体的敏感性。电子显微镜观察由D3 细胞培养物纯化的病原体形态。在酶联免疫吸附试验(ELISA)中,分别纯化病原体制作抗原,检测 1196份孕妇血清特异性抗体(IgG、IgM)。结果传代5-100代的D3细胞的染色体数均是96。丹参 (SMB,100 μg／ml)对D3细胞和病原体有明显的促增殖作用。D3细胞感染病原体后72 h,均出现明显的细胞病变效应(CPE)和荧光阳性细胞。电镜下见到了病原体典型的形态。结论永生性的D3细胞可用于培养病原体,所培养的病原体可制备用于诊断的抗原。
Objective To prepare＇the gene-transferred （D3）cells and to identify their susceptibility to several pathogens causing congenital infections. Methods The gene-transferred cells, D3 cells, were generated by transferring the DNA of human epidermoid earcinoma （HEP-2） cells into human embryonic lung （HEL） cells. The chromosomes of D3 cells were detected. The effect of Salvia Mihiorrhiza Bung（ SMB） on the replications of D3 cells and these pathogens were studied. The cytopathic effects in D3 cells infected with these pathogens were observed in microscope. The susceptibility of D3 cells to these pathogens was identified by indirect immunofluorescence assay （RFA）. The normal modalities of these purified pathogens from D3 cells culture were observed in electron microscope （EM）. The purified pathogens were manufactured as antigens to detect specific IgG and IgM in a total of 1196 pregnant women serum specimens by ELISA. Results The chromosome number of the D3 cell all was 96 when it had passed from 5th to 100th generation. The cytopathic effects （CPE） and the fluorescence positive cells were detected at 72 h post-infection with these pathogens. The normal forms of these purified pathogens from D3 cells culture were observed in EM. SMB can increase the proliferation of D3 cells and these pathogens. The optimum concentration of SMB is 100 μg/ml. Conclusion The findings suggest that the immortal D3 cells may be used in the culture of these pathogens that were used to prepare the antigen for diagnosis.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2006年第4期343-347,共5页
Chinese Journal of Microbiology and Immunology
基金
江苏省卫生厅(H9515)扬州市-扬州大学联合基金(2003039-7)
