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相關文獻
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頁籤選單縮合
題 名 | Detection of IgA Against Epstein-Barr Virus Bzlf-1 Replication Activator (ZEBRA) in Sera of Nasopharyngeal Carcinoma Patients with a Recombinant ZEBRA Protein=以合成之EB病毒ZEBRA蛋白偵測鼻咽癌病人血清內抗ZEBRA的IgA抗體 |
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作 者 | 許志宏; 杜宗陽; 林龍參; 羅麗華; 羅慕舜; 黃嘉嫻; 陳光耀; 劉武哲; | 書刊名 | 中華醫學雜誌 |
卷 期 | 62:6 1999.06[民88.06] |
頁 次 | 頁350-355 |
分類號 | 416.879 |
關鍵詞 | BZLF-1複製促動劑; Epstein-Barr病毒; 鼻咽癌; BZLF-1 replication activator; ZEBRA; Epstein-barr virus; Nasopharyngeal carcinoma; |
語 文 | 英文(English) |
英文摘要 | BACKGROUND: Epstein-Barr virus (EBV) is closely associated with nasopharyngeal carcinoma (NPC). An EBV-encoded immediate-early antigen, BZLF- 1 replication activator (ZEBRA) initiates EBV replication and expression in all NPC tumors. In this study, we investigated whether immunoglobulin A (IgA) against ZEBRA is present in the sera of patients with NPC, and whether it was able to be determined by enzyme-linked immunosorbent assay (ELISA) using a recombinant ZEBRA prepared from Escherichia coli. METHODS: A polymerase chain reaction-amplified cDNA fragment of the ZEBRA gene was inserted into the expression vector of E coli under the control of an IpL promoter. E coli bacteria containing the CI857 gene served as host to overexpress the ZEBRA protein by heat induction. Recombinant ZEBRA was collected by mechanical disruption of the bacteria, purified by column chromatography, and analyzed by SDS-PAGE and Western blot assay using sera from NPC patients. The recombinant ZEBRA was used to develop the ELISA to detect IgA against ZEBRA. RESULTS: The amount of ZEBRA produced comprised 30% of total E coli protein. Western blot assay confirmed that affinity of the recombinant ZEBRA to IgA antibody was preserved. IgA against ZEBRA was shown to be positive by ELISA in 36 of 40 NPC sera, but in only nine of 55 patients with other head and neck malignancies, and two of 35 normal individuals. For serologic diagnosis of NPC, the sensitivity of IgA/ZEBRA detected by ELISA was 90% and the specificity was 87.4%. CONCLUSIONS: A recombinant ZEBRA was produced at high levels in E coli and retained affinity to IgA against ZEBRA. The recombinant ZEBRA was successfully used to develop an ELISA for the detection of IgA against ZEBRA. The high sensitivity and specificity of IgA against ZEBRA show that the ELISA is feasible for serologic diagnosis of NPC. |
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