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頁籤選單縮合
題名 | 直接檢測Vibrio Vulnificus之聚合酶連鎖反應方法之開發=Development of the Polymerase Chain Reaction Method for Direct Detection of Vibrio Vulnificus |
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作者 | 吳美莉; 陳錦樹; 張敏政; | 書刊名 | 國立屏東科技大學學報 |
卷期 | 10:1 2001.03[民90.03] |
頁次 | 頁7-13 |
分類號 | 412.37 |
關鍵詞 | 聚合酶連鎖反應; 專一性; 靈敏度; Vibrio vulnificus; PCR; Specificity; Sensitivity; |
語文 | 中文(Chinese) |
中文摘要 | Vibrio vulnificus是一種會引發人類致命性感染的伺機性的革蘭氏陰性菌,本研究發展出一種利用聚合?連鎖反應快速檢測此種病原菌的方法。以vllY基因為目標序列,設計可以擴增452bp之DNA片段的專一性引子對(vP1及vP2)。在專一性檢測方面,自臨床檢體所分離的16株V. vulnificus皆可產生正反應。除此外,非Vibrio vulnificus之Vibri屬及非Vibrio屬之菌株皆無法呈現正反應。此聚合?連鎖反應系統可以至檢測100pg的染色體DNA,若配合1mM EDTA-0.5% Triton溶液之單一步驟萃取法進行模板DNA的萃取,其檢測靈敏度可達到102 CFU。 |
英文摘要 | Vibro vulnificus is an ubiquitous pathogen that is capable of causing a fatal infection in human. This study developed a PCR-based assay for the rapid identification of V. vulnificus. Specific primers (vP1 and vP2), which derved from vlly gene coding for V. vulnificus hemolysin, were designed for the amplification of 452 bp fragment within vlly gene specific ffor V. vulnificus. All 16 strains of v. vulnificus isolated from clinical specimens could generate positive results. In addition, Vibrio other than V. vulnificus and non-vibrio isolates did not yield positive results. Study on the detection sensitivity for this PCR system showed that the developed method was able to detect 100 pg of total genomic DNA. The PCR amplification, coupled with single-step DNA extraction method using a 1mM EDTA-0.5% Triton solution provided sensitivity limit to 102 CFU of V. vulnificus. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。