本文關(guān)鍵詞：基于環(huán)介導(dǎo)等溫?cái)U(kuò)增技術(shù)建立的假絲酵母菌快速檢測(cè)方法　出處：《中國(guó)現(xiàn)代醫(yī)學(xué)雜志》2016年24期 　論文類型：期刊論文

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【摘要】：目的基于環(huán)介導(dǎo)等溫?cái)U(kuò)增技術(shù)(LAMP)建立一種假絲酵母菌的快速檢測(cè)方法。方法依據(jù)假絲酵母屬18 S r DNA基因保守序列,采用Primer Explorer V4軟件設(shè)計(jì)引物,建立LAMP擴(kuò)增方法和反應(yīng)體系,對(duì)體系的4種關(guān)鍵參數(shù)和3項(xiàng)性能指標(biāo)進(jìn)行優(yōu)化和探討,并用臨床樣本進(jìn)行方法驗(yàn)證。結(jié)果 6種常見(jiàn)假絲酵母菌LAMP法檢測(cè)均為陽(yáng)性;最適擴(kuò)增條件為:溫度63~65℃、Bst酶活性4 IU、Mg2+濃度4 mol/L、擴(kuò)增時(shí)間1 h;7種對(duì)照菌種及人全血DNA樣本LAMP擴(kuò)增結(jié)果均為陰性;檢測(cè)限為102~107CFU/ml。經(jīng)52例臨床樣本驗(yàn)證,該法敏感性為100%,特異性為95%。結(jié)論該研究建立的LAMP假絲酵母菌檢測(cè)方法具有實(shí)用、簡(jiǎn)便、快速、特異和敏感的特點(diǎn),可為臨床提供一種新的假絲酵母菌診斷方法。
[Abstract]:Objective to establish a rapid detection method for Candida cerevisiae based on loop mediated isothermal amplification (Lampp). Methods the 18s r DNA gene of Candida was conserved. Primer Explorer V4 software was used to design primers, LAMP amplification method and reaction system were established, and four key parameters and three performance indexes of the system were optimized and discussed. Results all the six common Candida cerevisiae were positive by LAMP method. The optimum conditions for amplification were as follows: temperature 63 ~ 65 鈩，

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