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作 者:刘玉娟[1] 廖明凤[2] 易云兰[1] 张洁云[2] 杨倩婷[2] 张明霞[2]
机构地区:[1]深圳市第三人民医院妇产科,广东深圳518020 [2]深圳市第三人民医院广东省新发传染病诊治重点实验室,广东深圳518020
出 处:《中国热带医学》2015年第12期1420-1423,1435,共5页China Tropical Medicine
基 金:国家自然科学基金面上项目(No.81172732);广东省自然科学基金项目(No.2015A030313879);深圳市科技计划项目((No.JCYJ20130401164750004;JCYJ20140411111718168;JCYJ20150402111430653)
摘 要:目的建立盆腔积液结核分枝杆菌特异性IFN-γ酶联免疫斑点(PEMC Elispot)检测技术,并与相应的外周血检测结果比较。方法以重组结核菌早期分泌抗原6k Da蛋白(ESAT-6)、ESAT-6重组多肽抗原组合(P8.10)及多肽抗原组合(P8.10+Pool B)为抗原,建立盆腔积液结核分枝杆菌特异性IFN-γElispot检测技术(PEMC Elispot)。对46例确诊为结核性盆腔炎病人的盆腔积液进行结核菌特异性IFN-γ水平检测;同时用外周血结核菌特异性IFN-γ水平检测技术对病人相应的外周血进行检测,并用PEMC Elispot检测了30例非结核性盆腔炎病人积液标本。结果 PEMC Elispot技术对46例确诊为结核性盆腔炎病人盆腔积液检测,阳性率为97.8%;PBMC ELlispot检测病人外周血,阳性率为87.0%,经用四格表确切概率法统计,差异有统计学意义(χ2=6.815,P=0.009);PEMC Elispot检测非结核性盆腔炎病人盆腔积液的阳性率为10.0%,显著低于结核性盆腔炎病人盆腔积液阳性率(χ2=60.19,P<0.0001)。结论盆腔积液结核菌特异性IFN-γElispot检测技术与外周血结核杆菌特异性IFN-γElispot检测技术一样在结核性盆腔炎诊断应用中有着较好的敏感性和特异性,可以作为结核性盆腔炎诊断及其鉴别诊断的方法。Objective To develop Elispot assay for detection of Mycobacterium tuberculosis antigen-specific IFN-γ inpelvic tuberculousis(PEMC Elispot), and to compare it with Elispot assay for detection of Mycobacterium tuberculosisantigen-specific IFN-γ in corresponding peripheral blood(PBMC Elispot). Methods Recombinant early secretary antigentarget 6 k Da protein(ESAT-6)and the combination of ESAT-6 protein polypeptide(P8.10)、and P8.10+Pool B were used asantigens to develop the PEMC Elispot for measuring antigen specific IFN-γ in pelvic tuberculosis. IFN-γ response in pelvicfluid was detected with PEMC Elispot in 46 patients who were finally diagnosed as pelvic tuberculosis, and IFN-γ response incorresponding peripheral blood was determined simultaneously. Specimens of 30 cases non- tuberculous effusion in pelvicinflammatory disease were examined by PEMC Elispot. Results Pelvic fluid of 46 cases pelvic tuberculosis were tested byPEMC Elispot and the positive rate was 97.8%. Peripheral blood were detected by PBMC ELlispot and the positive rate was87.0%. There was significant difference between the results of PEMC Elispot and PBMC Elispot analyzed by the Fisher,exactprobability(χ^2=6.815,P=0.009). The positive IFN- γ response rate in pelvic tuberculosis fluid detected by PEMC Elispot(97.8%)was significantly higher than that of non- tuberculous pelvic fluid(10.0%)( χ^2=60.19,P〈0.0001). Conclusion PEMC Elispot as well as PBMC ELlispot in detection of Mycobacterium tuberculosis antigen-specific IFN-γ was with highsensitivity and specificity in diagnosis of pelvic tuberculosis. The PEMC Elispot assay can be applied to diagnosis anddifferential diagnosis of pelvic tuberculosis.
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