本文關(guān)鍵詞： 瘧疾 巢式PCR 卵形瘧原蟲 優(yōu)化應(yīng)用 敏感性 特異性　出處：《中國血吸蟲病防治雜志》2017年04期 　論文類型：期刊論文

【摘要】：目的結(jié)合瘧疾檢測工作實(shí)際,對瘧疾基因檢測的巢式PCR方法進(jìn)行優(yōu)化應(yīng)用,以提高瘧疾檢測效率。方法通過采用PCR預(yù)混液、內(nèi)引物一步擴(kuò)增、重新設(shè)計(jì)針對卵形瘧多個(gè)亞種的新引物,在瘧疾巢式PCR的基礎(chǔ)對反應(yīng)體系、反應(yīng)條件和卵形瘧原蟲特異性引物進(jìn)行優(yōu)化。對優(yōu)化后的方法進(jìn)行特異性和敏感性分析,并用巢式PCR和優(yōu)化后的方法同時(shí)對瘧疾陽性血樣和瘧疾送檢血樣進(jìn)行檢測,對檢測結(jié)果進(jìn)行比較分析。結(jié)果優(yōu)化后的方法特異性較好,敏感性可達(dá)pg至fg級。兩種方法同時(shí)檢測瘧疾陽性血樣的結(jié)果表明,優(yōu)化后的方法PCR擴(kuò)增產(chǎn)物的產(chǎn)量無明顯變化,非特異性擴(kuò)增明顯減少,卵形瘧不同亞種的檢出率提高,總體特異性提高。對111份瘧疾送檢血樣的實(shí)測結(jié)果表明,巢式PCR的敏感性和蟲種特異性分別為94.57%和86.96%,優(yōu)化后方法的敏感性和蟲種特異性均為93.48%,兩種方法敏感性差異無統(tǒng)計(jì)學(xué)意義(P0.05),但優(yōu)化后的方法特異性明顯提高(P0.05)。結(jié)論優(yōu)化后的PCR檢測在保持常規(guī)巢式PCR方法敏感性的基礎(chǔ)上提高了特異性,同時(shí)因其實(shí)驗(yàn)環(huán)節(jié)減少而節(jié)省了檢測成本、提高了檢測效率。
[Abstract]:Objective to optimize the application of nested PCR method for malaria gene detection in order to improve the efficiency of malaria detection. A new primer for multiple subspecies of Plasmodium ovum was redesigned to optimize the reaction system, reaction conditions and specific primers of Plasmodium ovum on the basis of malaria nested PCR. The specificity and sensitivity of the optimized method were analyzed. Nested PCR and optimized method were used to detect both malaria positive blood samples and malaria blood samples, and the results were compared and analyzed. The sensitivity of the two methods was from PG to FG. The results of the two methods showed that the yield of the PCR amplification products of the optimized method had no significant change, the non-specific amplification was significantly reduced, and the detection rate of different subspecies of oval malaria was increased. The overall specificity was improved. The measured results of 111 malaria blood samples showed that, The sensitivity and species specificity of nested PCR were 94.57% and 86.96, respectively. The sensitivity and species specificity of the optimized method were 93.48. There was no significant difference in sensitivity between the two methods (P 0.05), but the specificity of the optimized method was significantly improved. The optimized PCR detection improved the specificity on the basis of keeping the sensitivity of the conventional nested PCR method. At the same time, the cost of detection is saved and the efficiency of detection is improved because of the reduction of experimental links.
【作者單位】： 上海市疾病預(yù)防控制中心;
【分類號】：R531.3

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