Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/15309
標題: 以聚合酶鏈鎖反應及MSRV方法偵測豬隻屠體表面之沙門氏菌
Detection of Salmonella from pork carcass surface with a polymerase chain reaction based system and the MSRV method.
作者: 吳致銓
Wu, Chih-Chuan
關鍵字: MSRV
聚合酶
Salmonella
polymerase chain reaction
pork
carcass
鏈鎖反應
偵測
沙門氏菌

屠體
出版社: 獸醫病理學研究所
摘要: 在本研究中,主要是針對一種使用聚合酶鏈鎖反應做為核心技術的市售檢測套組 (BAX)、一種用Modified semi-solid Rappaport-Vassiliadis (MSRV)培養基之改良式快速培養法,以及USDA傳統培養法,以人工接種檢體方式與豬隻屠體表面擦拭檢體,進行測試。在靈敏度測試方面,BAX系統之PCR反應靈敏度達102 cfu/ml,而MSRV培養基則可達到100 cfu/ml。以人工接種檢體加以測試時發現,BAX法及MSRV法之靈敏度皆高於USDA傳統培養法10倍至100倍,兩方法都能在具有高量雜菌之檢體中檢出每ml僅數個之沙門氏菌菌體。在以長期冷凍乾燥沙門氏菌檢體所進行的平行測試中則顯示,直接MSRV法之靈敏度與穩定性遠優於USDA傳統培養法10倍至100倍。在使用230件豬隻屠體表面擦拭檢體之平行測試中,BAX法共檢出14件陽性檢體,而USDA傳統培養法則檢出包含上述14件中之8件陽性檢體。兩者之Kappa Cochen value為0.715,顯示BAX法與USDA傳統培養法間具有高度相關性。BAX法及MSRV快速培養法皆具有快速、高靈敏度及高穩定度的特性,在未來可望於篩檢大量檢體時取代USDA傳統培養法。
In this study, the BAX® for Screening/Salmonella, a commercial polymerase chain reaction based detection system, a Modified semi-solid Rappaport-Vassiliadis(MSRV) medium based rapid culture method and USDA traditional culture method were tested with artificial inoculated samples and pork carcass swab samples. The detection limit of BAX PCR reaction was 102 cfu/ml and 100 cfu/ml for MSRV medium. BAX method and MSRV method were 10 to 100 times more sensitive than USDA traditional culture method. Both of them could detect less than 10 cfu/ml salmonellae in artificial contaminated samples that presented more than 108 cfu/ml non-Salmonella florae. Direct MSRV method was 10 to 100 times more sensitive than USDA traditional culture method and showed with the highest sensitivity and stability in the long-term chilled salmonellae detection test. In the pork carcass sponge samples (n=230)screening test, the BAX method identified 14 samples as positive for Salmonella and the 8 samples identified as Salmonella-positive by the USDA traditional culture method were also BAX-positive. The Kappa Cochen value between the BAX method and the USDA traditional culture method is 0.715(substantial agreement). Both BAX method and MSRV method provide rapid screening alternative to the USDA traditional culture method for Salmonella detection.
URI: http://hdl.handle.net/11455/15309
Appears in Collections:獸醫病理生物學所

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