本文選題：日本血吸蟲 + 蟲卵�。� 參考：《中國寄生蟲學(xué)與寄生蟲病雜志》2017年03期

【摘要】：目的評估環(huán)介導(dǎo)等溫擴(kuò)增(loop mediated isothermal amplification,LAMP)法檢測糞樣中日本血吸蟲(Schistosoma japonicum)蟲卵DNA的效果,并評價其檢測流行區(qū)現(xiàn)場牛野外糞樣的效果。方法取1 g新鮮的日本血吸蟲蟲卵陰性牛糞,分別加入5個新鮮日本血吸蟲蟲卵、50μl日本血吸蟲蟲卵排泄分泌產(chǎn)物(egg secretion product,ESP)制備人工模擬陽性糞樣。用糞DNA提取試劑盒分別提取加入蟲卵的未經(jīng)研磨或研磨2 min后的人工模擬陽性糞樣、加入蟲卵ESP的糞樣和陰性糞樣的DNA,以日本血吸蟲28S核糖體DNA(r DNA)為檢測靶基因,用LAMP法、PCR法檢測,評估LAMP法檢測糞樣中日本血吸蟲蟲卵DNA的效果。收集2012-2014年湖南、湖北、江西、安徽等4省日本血吸蟲病流行區(qū)的牛野外糞樣221份,研磨后同法提取DNA,用LAMP法檢測,并與PCR法、孵化法的檢測結(jié)果進(jìn)行比較,評價其檢測流行區(qū)現(xiàn)場野外糞樣的效果。結(jié)果LAMP法檢測加入蟲卵糞樣并經(jīng)研磨后提取的DNA、加入蟲卵ESP糞樣提取的DNA均為陽性反應(yīng),呈綠色;而加入蟲卵糞樣未經(jīng)研磨提取的DNA、陰性糞樣DNA則為陰性反應(yīng),呈棕色;PCR檢測的結(jié)果與LAMP法檢測結(jié)果相近。LAMP法、PCR法和孵化法檢測血吸蟲病流行區(qū)牛野外糞樣的陽性率分別為5.43%(12/221)、4.52%(10/221)、0.90%(2/221)。LAMP法的陽性率明顯高于孵化法(P0.05),LAMP法與PCR法陽性率差異無統(tǒng)計學(xué)意義(P0.05)。結(jié)論 LAMP法可用于檢測流行區(qū)現(xiàn)場野外糞樣的日本血吸蟲蟲卵DNA,其現(xiàn)場應(yīng)用價值有待進(jìn)一步驗證。
[Abstract]:Objective to evaluate the effect of loop mediated isothermal amplification (LAMP) on the determination of the egg DNA of Schistosoma japonicum (Schistosoma japonicum) in the feces and to evaluate the effect of the detection of the field feces in the field of the epidemic area. Methods 1 g fresh Japanese Schistosoma japonicum egg negative cow dung was taken into 5 fresh Japanese blood respectively. An artificial simulated fecal sample was prepared by the egg excretory secretory product (egg secretion product, ESP) of 50 L Schistosoma japonicum. Using the fecal DNA extraction kit to extract the artificial simulated fecal samples that were added to the eggs without grinding or grinding 2 min, and adding the fecal and negative fecal samples of the eggs ESP to 28S ribosomes of Schistosoma japonicum. DNA (R DNA) was used to detect the target gene, LAMP method and PCR method to evaluate the effect of LAMP method for detecting the egg DNA of Schistosoma japonicum in the feces. 221 cattle field samples from 4 provinces of Japan schistosomiasis, Hunan, Jiangxi, Anhui and other provinces, were collected for 2012-2014 years, and DNA was extracted with the method of LAMP after grinding, and the detection results of the hatching method were with the PCR method. The results were compared. Results the LAMP method was used to detect the field feces in the field of the epidemic area. Results the DNA extracted from the eggs of the eggs was detected by the LAMP method, and the DNA of the fecal sample extracted from the eggs of the insect eggs was positive and green, and the negative fecal DNA was negative, and the negative fecal sample was brown; the PCR detection knot was detected by adding the eggs of the eggs ESP. The results were similar to that of LAMP method. The positive rate of PCR method and hatching method was 5.43% (12/221), 4.52% (10/221), and 0.90% (2/221).LAMP method was significantly higher than that of hatching (P0.05), and there was no statistical difference between LAMP method and PCR method (P0.05). Conclusion LAMP method can be used for detection flow. The field application value of field fecal DNA of Schistosoma japonicum remains to be further verified.

【作者單位】： 中國疾病預(yù)防控制中心寄生蟲病預(yù)防控制所世界衛(wèi)生組織熱帶病合作中心科技部國家級熱帶病國際聯(lián)合研究中心衛(wèi)生部寄生蟲病原與媒介生物學(xué)重點實驗室;湖南省血吸蟲病防治所;
【基金】：上海市衛(wèi)生和計劃生育委員會科研課題(No.201440498) 湖南省科技廳項目(No.2012FJ4103) 國家衛(wèi)生計生委疾控局委托項目(2015-2016) 中國疾控中心寄生蟲病所防治技術(shù)儲備科研基金(No.CB-15005)~~
【分類號】：R383.24

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