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肽酶联免疫吸附测定法和放射免疫沉淀测定法与体外翻译蛋白用于检测人乳头瘤病毒16型E6和E7蛋白血清抗体的比较

Comparison of peptide enzyme-linked immunosorbent assay and radioimmunoprecipitation assay with in vitro-translated proteins for detection of serum antibodies to human papillomavirus type 16 E6 and E7 proteins.

作者信息

Sun Y, Shah K V, Müller M, Muñoz N, Bosch X F, Viscidi R P

机构信息

Department of Immunology and Infectious Diseases, Johns Hopkins University School of Hygiene and Public Health, Baltimore, Maryland 21205.

出版信息

J Clin Microbiol. 1994 Sep;32(9):2216-20. doi: 10.1128/jcm.32.9.2216-2220.1994.

DOI:10.1128/jcm.32.9.2216-2220.1994
PMID:7529250
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC263970/
Abstract

Antibodies to human papilloma virus (HPV) type 16 (HPV-16) E6 and E7 proteins in serum are markers for HPV-associated invasive cervical carcinoma. We compared two assays, a radioimmunoprecipitation assay with in vitro-translated HPV-16 E6 and E7 proteins and an enzyme-linked immunosorbent assay (ELISA) with E6 and E7 synthetic peptides, for their abilities to discriminate serologically between patients with invasive cervical cancer and controls. Among the patients, antibody prevalences were higher by the E6 radioimmunoprecipitation assay (55.7%) than by the E6 peptide ELISA (15.5%), but among the controls, they were lower by the radioimmunoprecipitation assay (1.7%) than by the E6 peptide ELISA (5%). For E7, antibody prevalences among the patients were comparable by the radioimmunoprecipitation assay (43%) and the peptide ELISA (41%), but among the controls they were higher by the E7 peptide ELISA (17.4%) than by the radioimmunoprecipitation assay (4.1%). There was good agreement between the E7 radioimmunoprecipitation assay and the E7 peptide ELISA among patients but not among controls. In tests with representative sera, heat denaturation of the translated proteins resulted in a complete loss of reactivity to the E6 protein and a marked decrease in reactivity to the E7 protein. Our study showed that the radioimmunoprecipitation assay discriminates better than the peptide ELISA between patients with invasive cervical cancer and controls and that this is related to the ability of the radioimmunoprecipitation assay to detect conformational epitopes.

摘要

血清中针对人乳头瘤病毒16型(HPV - 16)E6和E7蛋白的抗体是HPV相关浸润性宫颈癌的标志物。我们比较了两种检测方法,一种是使用体外翻译的HPV - 16 E6和E7蛋白的放射免疫沉淀法,另一种是使用E6和E7合成肽的酶联免疫吸附测定法(ELISA),以评估它们在血清学上区分浸润性宫颈癌患者和对照组的能力。在患者中,E6放射免疫沉淀法检测到的抗体阳性率(55.7%)高于E6肽ELISA法(15.5%),但在对照组中,放射免疫沉淀法检测到的抗体阳性率(1.7%)低于E6肽ELISA法(5%)。对于E7,患者中放射免疫沉淀法检测到的抗体阳性率(43%)与肽ELISA法(41%)相当,但在对照组中,E7肽ELISA法检测到的抗体阳性率(17.4%)高于放射免疫沉淀法(4.1%)。患者中E7放射免疫沉淀法和E7肽ELISA法的检测结果一致性良好,但对照组中并非如此。在对代表性血清的检测中,翻译蛋白的热变性导致对E6蛋白的反应性完全丧失,对E7蛋白的反应性显著降低。我们的研究表明,放射免疫沉淀法在区分浸润性宫颈癌患者和对照组方面比肽ELISA法表现更好,这与放射免疫沉淀法检测构象表位的能力有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee70/263970/ecc4876cf889/jcm00009-0195-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee70/263970/75901b3f897f/jcm00009-0194-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee70/263970/ecc4876cf889/jcm00009-0195-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee70/263970/75901b3f897f/jcm00009-0194-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ee70/263970/ecc4876cf889/jcm00009-0195-a.jpg

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本文引用的文献

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Serologic response in human papillomavirus-associated invasive cervical cancer.人乳头瘤病毒相关浸润性宫颈癌的血清学反应
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