沙增管－沙測管套組（CMP Salmonella Enricher-Salmonella Checker Kit）篩檢生鮮肉品沙門氏菌的效能

Efficacy of the Salmonella Enricher-Salmonella Checker Kit for Screening Salmonella spp. in raw meat

陳昱樺(Yu Hua Chen) ； 歐柏廷(Po-Ting Ou) ； 蔡文城(Wen-cherng Tsai)

《檢驗及品保雜誌》 11卷1期 (2022/01) Pp. 14-19

https://doi.org/10.6573/JTQA.202201_11(1).03

摘要

傳統市場生鮮肉品（肉品或禽肉）為沙門氏菌的主要感染源，衛福部食品藥物管理署公告之食品中沙門氏菌（Salmonella spp.）檢驗方法（TFDA方法）操作甚為耗時、且需5－7天才能完成檢驗。本研究利用一種沙增管-沙測管套組（CMP Salmonella Enricher-Salmonella Checker kit）的高通量方法篩檢生鮮肉品中沙門氏菌，然後與TFDA方法檢測效能進行比較。操作TFDA方法時，將25g肉品檢體均質後，置入225 mL lactose broth進行增菌，在35℃隔夜培養後，取出少量培養液移種至Rappaport-Vassiliadis（R-V）增菌培養基，在42℃培養24±2小時後，再劃種至xylose lysine deoxycholate（XLD）agar。高通量方法則是利用一支人造絲（rayon-）或植絨拭子（flock-swab）塗抹適當面積（10×10cm^2）的肉品表皮，然後將拭子插入含Salmonella enricher的塑膠管（沙增管）中，在35℃培養8-17小時後，再取出拭子插入Salmonella checker的管（沙測管）中，在35℃培養24－48小時，若管內培養基出現黑色，則移出拭子劃線接種XLD平板培養基。上述兩種方法的XLD平板培養後，若出現疑似沙門氏菌的黑色菌落，則挑取單獨菌落劃種trypticsoyagar（TSA）平板，得到純菌後，再挑起單獨菌落利用matrix-assisted laser desorption ionization time-of-flight mass spectrometry（MALDI-TOFMS, Bruker biotyper, B-D公司，美國）以及傳統方法進行鑑定。結果指出TFDA方法與高通量方法分別顯示56%（14/25）與48%（12/25）的檢體帶有沙門氏菌，此指出傳統市場的禽畜肉品帶有高的沙門氏菌汙染率。雖然高通量方法（CMP Salmonella Enricher-Salmonella Checkerkit）的分離率稍低，但其操作簡易性、節省人力與物力，且可在短時間內檢測大量檢體，使其在篩檢肉品或屠體中的沙門氏菌污染時更具應用性。

關鍵字

TFDA方法 ； 沙增管－沙測管套組 ； 高通量方法 ； 沙門氏菌陽性分離率 ； 肉品或屠體

並列摘要

Raw meat (e.g., pork and poultry) sold in traditional markets is a major source of Salmonella infection. The method for detection of Salmonella spp. in food published by Taiwan Food and Drug Administration (TFDA) is very time-consuming and takes 5~7 days to complete. In this study, we evaluated the use of the Salmonella enricher- Salmonella checker kit for high-throughput screening of Salmonella spp. in raw meat and compared its efficacy with that of the TFDA method. For the TFDA method, a meat sample of 25 g was homogenized and placed in 225 mL of lactose broth for enrichment. After an overnight incubation at 35℃, a small aliquot of the inoculated broth was transferred to Rappaport- Vassiliadis (R-V) enrichment medium for 24±2 hours at 42℃ and then plated on xylose lysine deoxycholate (XLD) agar. For the high-throughput method, a rayon or flocked swab was used to collect sample by rubbing it on the surface (10×10 cm^2) of a piece of meat. The swab was then inserted into Salmonella enricher in a plastic tube and then transferred to Salmonella checker after 8~17 hours of incubation at 35℃. The swab was removed and streaked on an XLD agar plate if the Salmonella checker in the tube turned black after 24~48 hours of incubation at 35℃. For both methods, a colony suspected to be of Salmonella on XLD plates was picked and streaked on a tryptic soy agar (TSA) plate to obtain pure culture. Salmonella identification was then performed using both matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and conventional methods. Results showed that the positive Salmonella isolation rate by the TFDA method was 56% (14/25) compared to 48% (12/25) by the high-throughput method. Although the isolation rate of the high-throughput method is slightly lower, its ease of operation, labor and material savings, and ability to screen a large number of specimens in a short period of time would make it more applicable than the conventional method for monitoring Salmonella contaminations in raw meat or carcass.

並列關鍵字

TFDA method ； Salmonella enricher-Salmonella checker kit ； positive Salmonella isolation rate ； high-throughput method ； raw meat or carcass