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頁籤選單縮合
題 名 | 應用複合聚合酶連鎖反應檢測沙門氏桿菌=Detection of Salmonella by Multiplex Polymerase Chain Reaction |
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作 者 | 張惟茗; 廖明輝; 林敬覆; 黃春申; 郭靜蕙; 蕭終融; | 書刊名 | 行政院農業委員會家畜衛生試驗所研究報告 |
卷 期 | 41 民95.12 |
頁 次 | 頁147-152 |
分類號 | 437.255 |
關鍵詞 | 沙門氏桿菌; 複合聚合酶連鎖反應; 診斷; Salmonella; Multiplex polymerase chain reaction; Diagnosis; |
語 文 | 中文(Chinese) |
中文摘要 | 在獸醫、醫學或公共衛生環境保護等領域,沙門氏桿菌的檢測是一項重要但卻繁複的工作。傳統方法從檢體處理、培養至鑑定常需耗時3 至7 天才能完成,且相當耗費人力及材料。有鑑於此,本報告利用複合聚合酶連鎖反應(multiplex polymerase chain reaction, MPCR)快速及操作簡便的特點,進行invA及IS200基因檢測以輔助沙門氏桿菌之鑑定。本實驗共使用共263株分屬23種以上不同血清型之沙門氏桿菌,其中O 抗原有A、B、C1、C2、D1、D2、E1及E2群等,包括12株標準株及251株分離株進行測試。以MPCR檢測結果為除了2株H抗原歸類為z4 complex 檢測結果為陰性外,其他所有菌株之檢測結果均為陽性。450例豬糞便檢體中,以PCR檢驗有49例(10.89 %)為陽性。MPCR檢驗結果與傳統培養及鑑定法結果相符。傳統培養及鑑定法共分離到49株沙門氏桿菌分離株,其中包括12種不同血清型。50例鴨糞便檢體中,MPCR檢驗結果均為陰性,與傳統培養及鑑定結果相符。因此本方法確能有效檢出沙門氏桿菌,檢驗所需時間最長不超過18小時,可應用於沙門氏桿菌的快速輔助檢驗。 |
英文摘要 | A multiplex polymerase chain reaction (mPCR) assay, using two sets of primers designed according to the sequences of invA and IS200 genes, was developed for the identification of Salmonella. The mPCR assay was evaluated by using 12 seed strains and 251 isolated strains, which were attributed to more than 23 different serovars of group A, B, C1, C2, D1, D2, E1 and E2. All strain tested, except 2 possessed z4 complex (H antigen) strains, were positive in the mPCR assay. The mPCR assay was compared with the traditional culture and biological characteristics to determine the specificity and sensitivity of the test. Using a total of 450 swine fecal samples collected from slaughterhouse during 9 months, 49 samples were identified to be positive in both the mPCR assay and the traditional culture and biological characteristics. In addition, 50 duck fecal samples collected from animal house of duck were also tested, and all were found to be negative in both assays. Thus, the mPCR assay developed can be practically used as an auxiliary step for identifying Salmonella. |
本系統中英文摘要資訊取自各篇刊載內容。