| 陈吟霜 周帅 杨镒宇 谢永强 钟华敏 黄莲芬 姚淑雯 董慧 周珍文.金黄色葡萄球菌LukF-PV基因的原核表达及血清特异性抗体的检测[J].现代生物医学进展英文版,2015,15(23):4453-4456. |
| 金黄色葡萄球菌LukF-PV基因的原核表达及血清特异性抗体的检测 |
| Prokaryotic Expression and Detection of Staphylococcal Serum Specific Antibody of LukF-PV Gene |
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| DOI： |
| 中文关键词: 金黄色葡萄球菌LukF-PV 基因克隆 原核表达 血清特异性抗体 |
| 英文关键词: Staphylococcal LukF-PV gene Gene cloning Prokaryotic expression The antibody of serum |
| 基金项目:广州市医药科技重点项目（201102A212013） |
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| 中文摘要: |
| 目的：在大肠杆菌中表达金黄色葡萄球菌（Staphylococcus aureus , SA) LukF-PV 基因，纯化重组蛋白并以其为抗原检测儿童
SA 感染者血清特异性IgG 抗体，分析不同部位SA 感染患者血清LukF-PV 抗体水平。方法：将LukF-PV 克隆至pET-28a（+）载
体，IPTG 诱导重组蛋白表达、His 柱纯化重组蛋白后，用间接ELISA 检测儿童SA 感染者与健康对照者血清特异性IgG 抗体水
平。结果：成功表达纯化LukF-PV蛋白，儿童SA 感染组与健康对照组血清特异性抗体均值分别为（0.309± 0.063）、(0.505±
0.261)，具有统计学意义（P<0.05)。不同部位来源标本的感染者中血液组和扁桃体组抗体OD 均值分别为（0.634± 0.225）、
（0.481± 0.264）与健康对照组有显著差异（P<0.05)，其余各组与健康对照组无显著差异（P>0.05)。抗体阳性率统计分析中血液组分
别与脑脊液组、扁桃体组、咽拭子组、痰液组有统计学意义（P<0.05)，其余各组间均无显著差异（P>0.05)。结论：LukF-PV 成功表达
纯化,，儿童SA感染组LukF-PV特异性IgG 抗体显著高于健康对照组，其中来源于血液和扁桃体部位的SA 感染者抗体水平与
健康者有显著差异，尤以血液感染者最显著。 |
| 英文摘要: |
| Objective:To express Staphylococcus aureus LukF-PV gene in E.coli BL21, purify the recombinant protein, measure
and analyze serumspecific IgG antibody in children with different site SA infection by using recombinant LukF-PV as antigen.Methods:LukF-PV gene was cloned into pET-28a (+) vector, recombinant protein expression was induced by IPTG and recombinant protein was
purified by His colum, the SA infection patients' serum specific antibody level was measured by ELISA assay.Results:Recombinant
LukF-PV was successfully expressed and purified, the LukF-PV fusion protein was used as antigen for ELISA． The mean values of the
antibody levels of the subjects with SA-infected children group and the healthy control group were（0.309± 0.063）, (0.505± 0.261)
respectively (P<0.05)．The mean OD values of the antibody of the subjects with blood and tonsi were（0.634± 0.225）,（0.481± 0.264）,
respectively. There were significant differences compared with healthy control （P<0.05). while the other subjects showed no significant
difference with healthy control （P>0.05). Antibody positive rate of the subject blood showed significant difference with the subjects of
CSF, tonsil, throat swab, and sputum（P<0.05), while antibody positive rate of other subjects showed no significant difference（P>0.05).Conclusion:The recombiant LukF-PV protein was successfully expressed and purified, the LukF-PV -specific IgG antibody in SA
infection children was significantly higher than healthy control, especially in blood and tonsil patients, with the blood the highest . |
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