本文選題：嚴重急性呼吸綜合征相關(guān)冠狀病毒 + 纖突蛋白�。� 參考：《天津醫(yī)科大學(xué)》2006年碩士論文

【摘要】：嚴重急性呼吸綜合征(severe acute respiratory syndrome,SARS)又稱為傳染性非典型肺炎,是一種新出現(xiàn)的烈性傳染病。世界衛(wèi)生組織(WHO)已確認該病的病原體為一種新的冠狀病毒,命名為SARS冠狀病毒(SARS-CoV)。SARS疫病全球暴發(fā)給國際社會造成了極大的恐慌,給世界各國的衛(wèi)生事業(yè)以及經(jīng)濟發(fā)展帶來了嚴重后果。但是SARS在臨床上鑒別診斷比較困難,因此建立安全、特異、敏感的SARS-CoV抗原檢測方法對SARS的早期診斷、預(yù)防和治療具有重要意義。 本研究旨在制備抗基因工程表達的纖突蛋白(S)、核衣殼蛋白(N)的單克隆抗體(mAb),初步建立早期檢測SARS-CoV抗原的雙抗體夾心ELISA方法。我們以基因工程表達的S、N蛋白為免疫原,采用常規(guī)雜交瘤技術(shù)分別制備分泌抗S、N蛋白的mAb雜交瘤細胞株;對其進行鑒定,包括細胞克隆株穩(wěn)定性和染色體分析,檢測雜交瘤細胞培養(yǎng)上清及腹水中mAb效價、免疫球蛋白(Ig)分型,特異性分析,中和試驗,以及相關(guān)表位分析。以抗S蛋白、N蛋白抗體分別建立雙抗體夾心ELISA法檢測SARS-CoV抗原,對180份健康人血清、臨床診斷為SARS的12份抗體陰性血清和12份抗體陽性血清進行檢測。結(jié)果如下:成功制備出了3株分泌抗S蛋白mAb(B8G2,B10F6,C10G10)和2株分泌抗N蛋白mAb(F2G6G3,G5H12G12)的雜交瘤細胞株;雜交瘤細胞染色體計數(shù)接近兩種親本細胞染色體數(shù)目的總和;細胞培養(yǎng)上清和小鼠誘生腹水中mAb效價高;3株抗S蛋白mAb均為IgG1(κ),而抗N蛋白mAb G5H12G12為IgG2a(κ)、F2G6G3為IgG1(κ);單抗特
[Abstract]:Severe acute respiratory syndrome (SARS), also known as infectious atypical pneumonia, is a new severe infectious disease. The World Health Organization (WHO) has identified the pathogen of the disease as a new coronavirus named SARS coronavirus SARS-CoV. The global outbreak of SARS has caused great panic in the international community. For the world's health care and economic development has brought serious consequences. But the differential diagnosis of SARS is difficult in clinic, so it is important to establish a safe, specific and sensitive detection method of SARS-CoV antigen for early diagnosis, prevention and treatment of SARS. The aim of this study was to prepare a monoclonal antibody (mAba) against the genetically expressed plasminogen, nucleocapsid protein N, and to establish a sandwich ELISA method for the early detection of SARS-CoV antigens. The mAb hybridoma cell lines secreting anti-SfN protein were prepared by conventional hybridoma technique and identified, including cell clone stability and chromosome analysis. The mAb titers, immunoglobulin titers, specificity analysis, neutralization test and related epitope analysis were detected in the supernatant and ascites of hybridoma cells. Double antibody sandwich ELISA method was established to detect SARS-CoV antigen with anti S protein N protein antibody, and 12 antibody negative sera and 12 antibody positive sera were detected in 180 healthy people's sera. The results are as follows: three hybridoma cell lines secreting anti-S protein mAbB8G2OB10F6C10G10) and two hybridoma cell lines secreting anti-N protein mAb-F2G6G3 G5H12G12) were successfully prepared, and the chromosome number of hybridoma cells was close to the sum of the chromosomes of the two parents. The mAb titers of the supernatants of cell culture and the ascites of mice were all IgG1 (kappa) and IgG2a (魏 F2G6G3) for anti-S protein mAb, and IgG1 (kappa; monoclonal antibody) for anti-N protein mAb G5H12G12.
【學(xué)位授予單位】：天津醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R392

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