發(fā)布時(shí)間：2017-12-27 02:08

  本文關(guān)鍵詞：實(shí)時(shí)熒光定量PCR和PFGE在檢測(cè)海產(chǎn)品中副溶血性弧菌的應(yīng)用　出處：《現(xiàn)代預(yù)防醫(yī)學(xué)》2016年17期 　論文類型：期刊論文

  更多相關(guān)文章： 實(shí)時(shí)熒光定量PCR PFGE 海產(chǎn)品 副溶血性弧菌

【摘要】：目的應(yīng)用實(shí)時(shí)熒光定量PCR(RT-PCR)篩檢海產(chǎn)品中副溶血性弧菌,結(jié)合常規(guī)培養(yǎng)以提高檢出率;應(yīng)用PFGE對(duì)32株副溶血性弧菌進(jìn)行分子分型分析。方法在常規(guī)培養(yǎng)的基礎(chǔ)上,應(yīng)用RT-PCR對(duì)96件樣品增菌液進(jìn)行初篩,再用常規(guī)培養(yǎng)法確證陽(yáng)性;用陽(yáng)性似然比與陰性似然比、符合率與Kappa值做為指標(biāo),評(píng)價(jià)RT-PCR篩檢的真實(shí)性和可靠性;用限制性內(nèi)切酶SfiⅠ對(duì)32株菌進(jìn)行PFGE分子分型,用Bio Numerics Version 5.01軟件對(duì)圖像進(jìn)行聚類分析。結(jié)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(yáng)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(gè)PFGE圖譜類別,相似性在62.1%~100.0%之間,可分5個(gè)聚類群。結(jié)論 RT-PCR在檢測(cè)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(jià)值高;海產(chǎn)品中副溶血性弧菌親緣關(guān)系較遠(yuǎn),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(yù)防控制中心食物中毒診斷溯源技術(shù)北京市重點(diǎn)實(shí)驗(yàn)室;北京市東城區(qū)疾病預(yù)防控制中心;
【基金】：國(guó)家科技重大專項(xiàng)(2012ZX10004215-003-001) 北京市高層次衛(wèi)生技術(shù)人才培養(yǎng)計(jì)劃(2013-3-099)
【分類號(hào)】：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(xué)者從海魚中培養(yǎng)到的[3],截止到目前VP的檢測(cè)主要依賴傳統(tǒng)的常規(guī)培養(yǎng)法,也是公認(rèn)的“金標(biāo)準(zhǔn)”。該方法檢測(cè)能力有限

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