文章詳目資料

臺灣農業化學與食品科學 Scopus

  • 加入收藏
  • 下載文章
篇名 聚合酶連鎖反應技術檢測沙門氏桿菌之評估
卷期 48:3
並列篇名 Evaluation of the Polymerase Chain Reaction Technique for Detection of Salmonella
作者 王貞懿卓憲駿黃翠萍張育彰施養志
頁次 147-154
關鍵字 聚合酶連鎖反應沙門氏桿菌Polymerase chain reaction SalmonellaScopusTSCI
出刊日期 201006

中文摘要

以聚合酶連鎖反應 (PCR) 方法的四組引子檢驗沙門氏桿菌的三段基因 (sefA、invA、fmA) 及ITS區域,sefA為纖毛蛋白基因,invA為內膜蛋白基因,fimA為纖毛次單元的基因,ITS區域為16S和23S rDNA間的序列。藉由測試不同溫度條件對各組引子測定沙門氏桿菌DNA專一性的影響,進一步測試食品檢體,以比較不同引子的專一性。在沙門氏桿菌的測試中,invA及ITS引子的專一性測試結果為100% (95/95),fimA引子只有83% (79/95),而sefA引子專一性不佳。其檢測極限分別為invA引子1 ng、ITS引子100 pg、fmA引子1 ng;實際運用在食品檢體測試方面,invA及ITS引子對沙門氏桿菌檢出率亦均為100%,但ITS引子會檢測出非目標菌株,因此invA引子的專一性最佳,所以在本研究中invA引子適用於沙門氏桿菌的PCR快速檢測。

英文摘要

A polymerase chain reaction (PCR) method was used to detect Salmonella with 4 primer pairs to amplify 3 different genes (sefA, invA, fimA) and ITS regions. These 4 primer pairs amplify 4 different fragments included in (1) the sefA gene encoding the fmbrial protein, (2) the invA gene encoding an inner membrane protein, (3) the fmA gene encoding a major fmbrial subunit, and (4) the ITS region located between 16S and 23S rDNA. We studied whether different annealing temperatures affect the specifcity of the 4 primer pairs for detection of Salmonella DNA and compared the specifcity of the 4 primer pairs by testing food samples. The results for detection of Salmonella DNA showed that the invA and the ITS primers detected 100% (95/95) of the positive samples, the fmA primer detected 83% (79/95) of the positive samples, whereas the sefA primer was the least specific. The detection limits of this method were 1 ng, 100 pg and 1 ng for invA, ITS and fmA primers. In the results of testing food samples, the invA and ITS primers detected 100% of the positive samples, too. However, the ITS primer detected non-Salmonella. In this study, the invA primer is the most specifc for detection of Salmonella. It is concluded that using the invA primers, PCR could be a rapid and
reliable diagnostic tool for detection of Salmonella.

相關文獻