| 张彩勤张海赵勇毛峰峰赵善民杨巍白冰师长宏△.结核分枝杆菌分泌蛋白Ag85B、Hsp16.3和ESAT6在血清学检测中
的初步应用[J].,2011,11(3):401-404 |
| 结核分枝杆菌分泌蛋白Ag85B、Hsp16.3和ESAT6在血清学检测中
的初步应用 |
| Application of Ag85B,Hsp16.3 and ESAT6 of Mycobacterium Tuberculosisin Serological Detection |
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| DOI： |
| 中文关键词: 结核分枝杆菌 Ag85B Hsp16.3 ESAT6 酶联免疫吸附测定 |
| 英文关键词: Mycobacterium tuberculosis Ag85B Hsp16.3 ESAT6 ELISA |
| 基金项目:国家"863" 专项课题（2007AA02Z473）；国家自然科学基金（30972767）；陕西省自然科学基金（SJ08C203）联合资助 |
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| 中文摘要: |
| 目的：探讨结核分枝杆菌分泌蛋白Hsp16.3、Ag85B 以及融合蛋白ESAT6-CFP10、Ag85B-Hsp16.3 和Ag85B-ESAT6 用于
TB病人血清学检测的意义。方法：将已构建的含5种目的基因的表达载体（pProEXHTb-Hsp16.3、pProEXHTa-Ag85B、pProEXHTb-
ESAT6-CFP10、pProEXHTa-Ag85B-Hsp16.3、pProEXHTa-Ag85B-ESAT6），分别转入宿主菌E.coli DH5α中，诱导表达后分别获得
Hsp16.3、Ag85B、ESAT6-CFP10、Ag85B-Hsp16.3 和Ag85B-ESAT6 五种蛋白，采用Ni2+亲和层析柱进行纯化，并用透析方法进行
目的蛋白的复性。将经过复性的5种蛋白分别作为抗原，采用间接ELISA方法检测待测的血清样本，经OPD 显色，测定各孔
OD490值并判定结果。结果：五种蛋白被成功纯化并复性，通过ELISA 方法共检测了22 例TB 病人血清、10 例非结核病人血清
和6 例正常对照血清，Hsp16.3、Ag85B、ESAT6-CFP10、Ag85B-Hsp16.3 和Ag85B-ESAT6 这5 种抗原的灵敏度分别为36.4 %、
90.9 %、77.3 %、95.5 %、100 %，特异性分别为100 %、75 %、100 %、93.8 %、93.8 %。统计分析显示，ESAT6-CFP10 和Ag85B、
Ag85B-Hsp16.3、Ag85B-ESAT6 这三种蛋白ELISA 检测的结果无差异，而与Hsp16.3 和痰涂片检测结果有显著差异。结论：
Ag85B-Hsp16.3和Ag85B-ESAT6可作为结核分枝杆菌ELISA检测的初选抗原。 |
| 英文摘要: |
| Objective: To investigate the significance of Hsp16.3, Ag85B, ESAT6-CFP10, Ag85B-Hsp16.3 and Ag85B-ESAT6 of
Mycobacterium tuberculosis (MTB) in the serological detection. Methods: The five recombinant plasmids (pProEXHTb-Hsp16.3,
pProEXHTa-Ag85B, pProEXHTb-ESAT6-CFP10, pProEXHTa-Ag85B-Hsp16.3 and pProEXHTa-Ag85B-ESAT6) were transfered into
E.coli DH5α, respectively. The five proteins were induced by IPTG and purified by Ni-NTA purification system under denaturing condition.
Following by renaturation of dialysis, the five proteins were used as antigen to detect sera of 22 TB patients, 10 non-TB patients and
6 controls by indirect ELISA. OPD was chromogenic agent, and OD490 were detected. Results: The sensitivities of Hsp16.3, Ag85B,
ESAT6-CFP10, Ag85B-Hsp16.3 and Ag85B-ESAT6 to 22 TB patients were 36.4 %, 90.9 %, 77.3 %, 95.5 %, 100%, respectively, while
the specificity to 16 non-TB patients were 100 %,75 %,100 %,93.8 %,93.8 %, respectively. There was no difference for the ELISA detection
result between ESAT6-CFP10 and Ag85B, Ag85B-Hsp16.3, Ag85B-ESAT6, while there was difference between ESAT6-CFP10
with Hsp16.3 and sputum-smear. Conclusion: Ag85B-Hsp16.3 and Ag85B-ESAT6 could be used as the original selective antigens in detecting
MTB by ELISA. |
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