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題 名 | Use of Polymerase Chain Reaction-Restriction Fragment Analysis for Detection and Identification of Mycobacterium Tuberculosis=利用核酸聚合酵素連鎖反應與限制酵素分解片斷分析方法以檢測及鑑定結核分枝桿菌 |
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作 者 | 鄭乃琳; 林依靜; 魏誠佑; 曹汶龍; 黃娟娟; 李展平; | 書刊名 | 慈濟醫學 |
卷 期 | 11:4 1999.12[民88.12] |
頁 次 | 頁311-320 |
分類號 | 415.12 |
關鍵詞 | 分子診斷; 結核分枝桿菌; 核酸聚合酵素連鎖反應; 限制酵素分解片段分析; Molecular diagnosis; Mycobacterium tuberculosis; PCR; Restriction fragment analysis; |
語 文 | 英文(English) |
英文摘要 | Objective: We combined the PCR assays developed previously with restriction fragment analysis (PCR-RFA) to detect and identify Mycohacterium tuberculosis. Patients and Methods: The PCR assay itself targets three DNA: the 65 kDa antigen protein gene for detecting a wide spectrum ofmycobacterial strains; the IS6110 insertion sequence for detecting the My cobacterium tuberculosis complex, mainly M. tuberculosis and M. bovis BCG; and the mtp40 genomic fragment for distinguishing M. tuberculosis from M. bovis BCG. RFA, on the other hand, requiring no DNA sequencing or radioactive hybridization, plays a confirmation role to ensure the correctness of the amplified DNA products. Results: Several restriction enzymes, with properties of unique cutting site at the asymmetrical location of the expected amplicon and simple digestion conditions, have been selected and tested in this method. Among these. Sac a is suitable for both of the ampliconsfrom the IS6110 and the mtp40 targets, whereas Eco 01091 is suitable for the amplicon from the 65 kDa target. Conclusions: Taken together, this PCR-RFA method has been used successfully in the detection and identification ofM. tuberculosis in a CSF sample from a patient with tuberculous meningitis. |
本系統中英文摘要資訊取自各篇刊載內容。