本文關鍵詞：實時熒光定量PCR和PFGE在檢測海產(chǎn)品中副溶血性弧菌的應用　出處：《現(xiàn)代預防醫(yī)學》2016年17期 　論文類型：期刊論文

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【摘要】：目的應用實時熒光定量PCR(RT-PCR)篩檢海產(chǎn)品中副溶血性弧菌,結合常規(guī)培養(yǎng)以提高檢出率;應用PFGE對32株副溶血性弧菌進行分子分型分析。方法在常規(guī)培養(yǎng)的基礎上,應用RT-PCR對96件樣品增菌液進行初篩,再用常規(guī)培養(yǎng)法確證陽性;用陽性似然比與陰性似然比、符合率與Kappa值做為指標,評價RT-PCR篩檢的真實性和可靠性;用限制性內(nèi)切酶SfiⅠ對32株菌進行PFGE分子分型,用Bio Numerics Version 5.01軟件對圖像進行聚類分析。結果 96件樣品常規(guī)培養(yǎng)法檢出率24.00%,RT-PCR/常規(guī)培養(yǎng)法檢出率33.33%。陽性似然比為3.88,陰性似然比為0.06。符合率為80.21%,中、高度一致(Kappa值=0.57);32株菌得到31個PFGE圖譜類別,相似性在62.1%~100.0%之間,可分5個聚類群。結論 RT-PCR在檢測海產(chǎn)品中副溶血性弧菌有很好的初篩作用,RT-PCR/常規(guī)培養(yǎng)法使用價值高;海產(chǎn)品中副溶血性弧菌親緣關系較遠,顯示高度多態(tài)性。
[Abstract]:Objective to detect Vibrio parahaemolyticus in seafood by real-time fluorescence quantitative PCR (RT-PCR), and to improve the detection rate by routine culture. 32 strains of Vibrio parahaemolyticus were analyzed by PFGE. Methods on the basis of routine training on the application of RT-PCR in 96 samples of broth were screened, and the conventional culture method confirmed positive; positive likelihood ratio and negative likelihood ratio, the coincidence rate as the index and Kappa value, authenticity and reliability evaluation of RT-PCR screening; 32 strains were PFGE molecular typing with the restriction endonuclease Sfi, cluster analysis was used to detect Bio Numerics Version 5.01 software. Results the detection rate of the conventional culture of 96 samples was 24%, and the detection rate of the conventional culture method of RT-PCR/ was 33.33%. The positive likelihood ratio was 3.88, and the negative likelihood ratio was 0.06. The coincidence rate was 80.21%, middle and high (Kappa value =0.57); 32 strains obtained 31 PFGE map categories, similar to 62.1%~100.0%, can be divided into 5 cluster groups. Conclusion RT-PCR has a good initial screening effect in detecting Vibrio parahaemolyticus in seafood. RT-PCR/ has a high value in routine culture. The Vibrio parahaemolyticus in seafood is far away from each other, showing high polymorphism.
【作者單位】： 北京市疾病預防控制中心食物中毒診斷溯源技術北京市重點實驗室;北京市東城區(qū)疾病預防控制中心;
【基金】：國家科技重大專項(2012ZX10004215-003-001) 北京市高層次衛(wèi)生技術人才培養(yǎng)計劃(2013-3-099)
【分類號】：R440;R155.5
【正文快照】： 副溶血性弧菌(Vibrio parahaemolyticus,VP)屬于弧菌科弧菌屬,廣泛存在于海產(chǎn)品中,是引起感染性腹瀉和食源性疾患的重要病菌[1-2]。VP的首次檢出就是1953年日本學者從海魚中培養(yǎng)到的[3],截止到目前VP的檢測主要依賴傳統(tǒng)的常規(guī)培養(yǎng)法,也是公認的“金標準”。該方法檢測能力有限

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