沙眼衣原体感染免疫优势蛋白的血清学筛选

Screening of immunodominant proteins for sensitive detection of Chlamydia trachomatis infection by serological test

目的：检测泌尿生殖道沙眼衣原体感染者血清中抗重组衣原体质粒编码蛋白3（rPgp3）、衣原体蛋白酶样活性因子（ rCPAF）、Ct143编码蛋白（ rCT143）、Ct101编码蛋白（ rCT101）、Ct694编码蛋白（ rCT694）、Ct875编码蛋白（ rCT875）、Ct813编码蛋白（ rCT813）、主要外膜蛋白（rMOMP）、膜蛋白 A（rIncA）和热休克蛋白60（rHsp60）的 IgG 抗体，比较上述几种蛋白的抗原性。方法将编码上述10种蛋白的基因重组质粒转化入大肠杆菌，经 IPTG 诱导表达并鉴定后，用裂解液包被谷胱甘肽预包被的 ELISA 孔板，饱和板底的谷胱甘肽。收集天津医科大学总医院性病门诊经胶体金法和单克隆抗体免疫荧光法检测沙眼衣原体两者均阳性者的血清50份和两者均阴性者的血清10份，进行 ELISA 法检测。结果50份沙眼衣原体感染者血清中 rPgp3、rCPAF、rCT143、rCT101、rCT694、rCT875、rCT813、rMOMP、rIncA、rHsp60蛋白抗体的检出率分别是44例（88％）、38例（76％）、37例（74％）、36例（72％）、33例（66％）、31例（62％）、30例（60％）、26例（52％）、24例（48％）、17例（34％）。10份未感染者血清未检测到上述蛋白的抗体。结论上述10种沙眼衣原体抗原蛋白中，rPgp3抗原性最强，抗体检出率最高，其次是 rCPAF 和 rCT143；rHsp60抗原性最弱，抗体检出率最低。

Objective To detect the antibodies against recombinant chlamydial plasmid-encoded protein 3（rPgp3）,chlamydial protease-like activity factor（rCPAF）,Ct143 encoded protein（rCT143）, Ct101 encoded protein（rCT101）,Ct694 encoded protein（rCT694）,Ct813 encoded protein（rCT813）, Chlamydia membrane protein A（rIncA）,Ct875 encoded protein（rCT875）,major outer membrane protein （rMOMP）and heat shock protein 60（ rHsp60）in serum samples collected form patients with urogenital Chlamydia trachomatis（Ct）infection and to evaluate the antigenicity of those proteins. Methods The recombinant plasmids expressing the 10 proteins and a blank plasmid were transformed into E. coli BL21 strains,respectively. The transformed E. coli BL21 strains were induced by isopropyl β-D-1-thiogalactopyr-anoside（IPTG）to express recombinant proteins. The glutathione pre-coated 96-well ELISA plates were coa-ted with lysates. Serum samples were collected from 50 patients with Ct infection and 10 patients without Ct infection. ELISA was performed to detect the antibodies against 10 recombinant proteins. Results The anti-bodies against rPgp3,rCPAF,rCT143,rCT101,rCT694,rCT875,rCT813,rMOMP,rIncA and rHsp60 proteins were respectively detected in 44 cases（88% ）,38 cases（76% ）,37 cases（74% ）,36 cases （72% ）,33 cases（66% ）,31 cases（62% ）,30 cases（60% ）,26 cases（52% ）,24 cases（48% ）and 17 cases（34% ）out of 50 serum samples. No antibodies against 10 recombinant proteins were detected in the serum samples collected from patients without Ct infection. Conclusion The rPgp3 protein showed the strongest antigenicity among all of the studied proteins,followed by rCPAF and rCT143 proteins. The rHsp60 protein showed the lowest antigenicity.