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題 名 | Comparison of the Sensitivity and Specificity of an Automatic Ligase Chain Reaction Assay System with a One-Step Polymerase Chain Reaction Assay in the Diagnosis of Mycobacterium Tuberculosis Complex=比較自動化連接酵素與單一步驟聚合酵素連鎖反應方法應用在診斷結核分枝桿菌的敏感性與專一性 |
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作 者 | 林依靜; 謝閔傑; 葉政憲; 黃娟娟; 魏誠佑; 曹汶龍; 李展平; | 書刊名 | 長庚醫學 |
卷 期 | 22:2 1999.06[民88.06] |
頁 次 | 頁204-211 |
分類號 | 415.12 |
關鍵詞 | 連接酵素連鎖反應; 分子診斷; 結核分枝桿菌; 聚合酵素連鎖反應; Ligase chain reaction; LCR; Molecular diagnosis; Mycobacterium tuberculosis; Polymerase chain reaction; PCR; |
語 文 | 英文(English) |
中文摘要 | 背景:聚合酵素連鎖反應和連接酵素連鎖反應為兩種核酸增殖放大原理的分子診 斷方法,前者已被廣泛使用在結核分枝桿菌的分子診斷,而後者對我們醫學界而言是較陌生 的。為了評估何種核酸增殖放大方法較適合用在結核分枝桿菌複合群的分子診斷,我們比較 這兩種方法的敏感度及專一性。 分法:我們比較一套自動化連接酵素連鎖反應 (automatic LCR) 及單一步驟聚合酵素連鎖反應 (one-step PCR) 在結核分枝桿菌的分子診斷上的表現 。 由亞培公司所研發出的自動化 LCR 是以單一數目的抗原蛋白乙基因作為標的核酸;而單 一步驟 PCR 方法則利用多重複的 IS6110 核酸插入序列作為其標的核酸, 此兩種方法對於 結核分枝桿菌複合群都具有專一性。結果:我們的研究結果顯示兩種方法都可檢測到結核分 枝桿菌複合群中的結核分枝桿菌及牛型分枝桿菌,而不會失誤檢測到其它的分枝桿菌菌株。 此外, 這兩種不同的核酸增殖放大技術都有相當好的敏感度:自動化 LCR 與單一步驟 PCR 方法可以分別敏感的檢測到 10 個及 100 個結核分枝桿菌基因體。 然而當標的核酸數目低 時,自動化 LCR 方法比單一步驟 PCR 方法的重複性較低。結論:我們的研究結果顯示除了 PCR 方法外,LCR 方法也是診斷結核分枝桿菌複合群的另一種有用的分子診斷方法,唯自動 化儀器的重複性部份須加強。 |
英文摘要 | Background:Polymerase chain reaction (PCR) and ligase chain reaction (LCR) are two nucleic acid amplification-based molecular methods. The former has been used widely in the identification of Mycobacterium tuberculosis (M. tuberculosis). In contrast, the LCR assay which was recently introduced is not well known in our medical communities in Taiwan. To determine which method is more reliable and suitable for the identification of M. tuberculosis in our clinics, we compared the sensitivity and specificity of these two methods. Methods:An automatic LCR assay system and a manual one-step PCR assay were studied in a side by side comparison of their performance in detection of M. tuberculosis. The automatic LCR system uses the single copy antigen protein b (Pab) gene and the manual one-step PCR assay uses the multi-copy IS6100 insertion element as the target DNA; both target DNA sequences are found specifically in M. tuberculosis complex. Results:Both assays detected two of the M. tuberculosis complex strains, M. tuberculosis and M. bovis, but not other mycobacterial strains. In addition, both methods, which were based on different amplification principles, showed compatible sensitivity; as low as 10 and 100 copies of M. tuberculosis genomes were detected by the LCR and PCR assays, respectively. When the template DNA was less than 1000 copies, however, the automatic LCR assay system showed a lower reproducibility than that of the one-step PCR assay. Conclusion:Our results suggest that in addition to the PCR assay, the LCR assay is a useful method for the molecular identification of M. tuberculosis complex strains. |
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