評估CMP efSwab-CB Broth, efSwab-GN Broth與efSwab-SBG Broth應用於糞便檢體沙門氏菌及／與志賀氏菌之保存／增菌效能

Evaluation of efSwab-CB Broth, efSwab-GN Broth and efSwab-SBG Broth for Preservation and Enrichment of Salmonella and Shigella in Fecal Specimens

胡郁旋(Yu-Hsuan Hu) ； 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. 7-13

https://doi.org/10.6573/JTQA.202201_11(1).02

摘要

從糞便檢體偵測沙門氏菌（Salmonella spp.）及志賀氏菌（Shigella spp.）通常以棉花或人造絲拭子採檢後置入含有非營養性Cary Blair（CB）semi-solid medium之半固態嗜氧輸送管中輸送至檢驗室，然後再接種適當平板培養基及一種增菌培養基。本研究，吾等評估由啟新生物股份有限公司製造的三種輸送培養基，包括CMP efSwab-CB（Cary Blair）broth, efSwab-GN（Gram negative）broth,與efSwab-SBG（selenite brilliant green sulfa）broth，用於偵測沙門氏菌與志賀氏菌的效能。每一種套組搭配含一種稱為efSwab之高吸附性及釋放能力的植絨拭子。此些輸送培養基可應用於自動化微生物檢驗室之自動化接種／劃片機台如Kiestra InoqulA（BD）或WASP（Copan）。試驗時，分別以沙門氏菌與／或志賀氏菌的純菌以及其與糞便混合的模擬檢體分別接種每一種輸送培養基，然後存放於室溫（26℃）與冷藏（4℃）不同時間（6、24與48h），結果顯示（i）兩種純菌在efSwab-CB broth以4℃保存，兩菌約可維持原菌量，但26℃保存，沙門氏菌可維持原菌量，而志賀氏菌的菌量則隨著存放時間而降低菌數。；（ii）含有沙門氏菌的糞便檢體在efSwab-CB broth無論存放於4℃或26℃，僅能於24小時內檢出，48小時則否；而含有志賀氏菌者，存放4℃僅能在6小時檢出，其餘存放時間則否，而於26℃，任何存放時間均不能檢出。（iii）沙門氏菌純菌在CMP efSwab-GN broth與efSwab-SBG broth以及志賀氏菌純菌在CMP efSwab-GN broth於4℃，菌數將隨著時間而大幅降低；而於26℃，存放6、24h與48h，沙門氏菌純菌在CMP efSwab-GN broth，菌數分別增加約8、2,303與2,674倍，而在CMP efSwab-SBG broth，菌數分別增加約4、488與206倍；志賀氏菌純菌於CMP efSwab-GN broth則分別增加菌量約2、1,259與4,553倍。（iv）含有沙門氏菌的糞便檢體在efSwab-GN broth與efSwab-SBG broth中不論存放於4℃或26℃，沙門氏菌於48小時內均能被檢出；以及（v）含有志賀氏菌的糞便檢體，志賀氏菌在4℃只能存活至24小時，而在26℃則僅存活至6小時。綜合上述，檢驗室使用自動劃片機，欲分離糞便中的非傷寒性沙門氏菌最好使用CMP efSwab-SBG broth套組在26℃或較高溫度（如35℃）存放，且於6－24小時間移種；而志賀氏菌則使用CMP efSwab-GN broth套組，且在同樣溫度存放但於4～6小時內移種。從糞便中分離此兩菌均不建議使用CMP efSwab-CB broth套組。

關鍵字

沙門氏菌 ； 志賀氏菌 ； CMP efSwab-CB（Cary Blair）broth ； CMP efSwab-GN broth ； CMP efSwab-SBG（selenite brilliant green sulfa）broth

並列摘要

For detection of Salmonella spp. and Shigella spp. in fecal specimens, a cotton or rayon swab is usually used to collect specimens. The swab is then placed in an aerobic transport tube containing non-nutritive Cary Blair( CB)semi-solid medium, transport to a laboratory and then inoculate on appropriate agar plates or in an enrichment medium. In this study, we evaluated the efficacy of three kinds of transport media manufactured by Creative Biotechnology Co., Inc. in Taiwan, including CMP efSwab-CB (Cary Blair) broth, efSwab-GN (Gram negative) broth, and efSwab-SBG (selenite brilliant green sulfa) broth for detection of Salmonella spp. and Shigella spp. Each kit includes a high adsorption and release capacity flocking swab, referred to as efSwab. These transport media can be used in microbiology laboratories with automated plating/streaking instruments such as InoqulA(BD) or WASP(Copan). For the tests, each transport medium was inoculated with a pure Salmonella or Shigella culture or a fecal sample spiked with Salmonella or Shigella. The inoculated transport media were stored at room temperature (26℃) and 4℃ for 6, 24, or 48 hours. Results showed the following: (i) The number of Salmonella and Shigella pure culture in efSwab-CB broth was decreased over time at 4℃ or 26℃; (ii) The Salmonella in spiked stool specimens placed in efSwab-CB broth and stored at 4℃ or 26℃ could only be detected within 24 hours, and the Shigella in spiked stool specimens placed in the same medium could only be detected within 6 hours of storage at 4℃; (iii) The number of Salmonella pure culture in CMP efSwab-GN broth or in CMP efSwab-SBG broth and that of Shigella pure culture in efSwab- GN broth was decreased over time at 4℃; When the specimens were stored at 26℃, the number of Salmonella pure culture in CMP efSwab-GN broth was increased 8, 2,303, and 2,674 times and that of those in CMP efSwab-SBG broth was increased 4, 488, and 206 times, whereas that of Shigella pure culture placed in efSwab-GN broth was increased 2, 1259, and 4,553 times, respectively, after 6, 24, and 48 h of storage; (iv) The Salmonella in spiked fecal specimens was detectable in efSwab-GN broth or in efSwab-SBG broth within 48 hours of storage at 4℃ or 26℃; (v)The Shigella in spiked stool specimens survived only up to 24 hours at 4℃ and up to 6 hours at 26℃. In conclusion, the best way to isolate non-typhoid Salmonella from stool specimens is to use CMP efSwab-SBG broth as the transport medium, and the specimen in the transport medium stored at 26℃ or higher (e.g., 35℃) is inoculated in appropriate media in 6-24 hours after collection. For Shigella isolation, the fecal specimens should be transported in CMP efSwab-GN broth and sub-cultured within 4~6 hours. The efSwab-CB broth is not recommended for the isolation of either Salmonella or Shigella from stool specimens.

並列關鍵字

Salmonella spp ； Shigella spp ； CMP efSwab-CB (Cary Blair) broth ； CMP efSwab-GN broth ； CMP efSwab-SBG (selenite brilliant green sulfa)broth