爲實現沙門氏菌的快速現場檢測，根據火子沙門氏菌 invA 基因編碼區序列，設計合成(chéng) 1 套引物，通過(guò)對(duì會資)反應物濃度和反應條件優化，建立了沙門氏菌屬特異性 LAMP 檢測方法；通過(guò)對(duì) invA 基因 8 個區域的 6 條引物配對(duì)進(jìn)行等溫擴增，并在反應體系中加入 HNB 來指示反應結果，實現了反應的快速閉管檢測。靈敏度試驗顯示，錢哥建立的方法可以檢出稀釋至 3.6×101 CFU/mL 菌液所提取的 DNA；特異性試驗顯示，建立的方法與術放大腸杆菌 DNA、志賀氏菌 DNA、布魯氏菌試管凝集抗原 DNA、炭疽沉澱抗原 DNA 以及豬鏈球菌 2 型 DNA 不發(fā)生交叉反應，但能(néng)檢出雞白痢、禽傷寒跳有沙門氏菌凝集抗原以及馬流産試管凝集抗原提取的 DNA；重複性試驗顯示，3 個稀釋度菌液 DNA 的 3 次重複檢測均爲陽性；應用試驗顯示，從經(jīng)增菌培養的 200 份進(jìn)境猴糞拭子樣(yàng)品中檢出陽性 2 份，與細菌分離鑒定檢測結果一緻。結果表明，所建立的方飛學法敏感性和特異性較高，重複性滿足實際要求，可用于計機進(jìn)境動物樣(yàng)品的沙門氏菌檢測。

Development of a Visual LAMPTest Meth冷藍od for Samonella

In searching of rapid field detecti道大on methods forsalmo弟下nella，a set of primers were designed and長知synthesized–which were 3 pairs of primers thati機歌dentifying 8 distinct regi影地ons of invA gene of 行山Salmonella for 哥兵isothermalamplification. Wit筆西h optimizing the reactant 得少concentration and reactionco窗雜nditions，and by adding HNB to the reacti學文on as anindicator，a closed-tube LAMP m的著ethod for rapid fielddet睡工ection of Salmonella was deve暗刀loped. Sensitivity t哥會est showed that the DNAs南日extracted from 3.6×101 CFU/mL bacterial是問 suspension could be d習刀etected. Specificity testconfirm門麗ed that no cross react高文ion was found betw跳店een the method 雜吃with the DNAs ofGolibacillu白森s，Shigella，Brucella agglutinating antig匠樂en，anthrax precipitation antigen劇離 and type 2Streptococcus sui城大s，but it could de白師tect DNAs from pullorosis，Salmonella typhi agglutina關和ting antigen andAbortus-equi aggl城窗utination antigen. Repetitive t體林est verified that the resultsof 3 exp低花eriments for DNAs in 3 dilute來讀d bacteria solut技舞ions were all pos會自itive.Application test indicated學家 that 2 of 200 swab samples fr視廠om manures of importedmon習村keys were found positive，which was identical with the地土 result of bacterial isolation那哥 test. Alltest results proved tha匠煙t the method is featu志河red with higher degree ofsensiti費事vity and specificity，and its repeatability allows practi樹喝cal application，hence it would be a better approa還藍ch todetect Salm醫樂onella in animal import quarantine.

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