本文選題：日本血吸蟲(chóng) + 蟲(chóng)卵　； 參考：《中國(guó)寄生蟲(chóng)學(xué)與寄生蟲(chóng)病雜志》2017年03期

【摘要】：目的評(píng)估環(huán)介導(dǎo)等溫?cái)U(kuò)增(loop mediated isothermal amplification,LAMP)法檢測(cè)糞樣中日本血吸蟲(chóng)(Schistosoma japonicum)蟲(chóng)卵DNA的效果,并評(píng)價(jià)其檢測(cè)流行區(qū)現(xiàn)場(chǎng)牛野外糞樣的效果。方法取1 g新鮮的日本血吸蟲(chóng)蟲(chóng)卵陰性牛糞,分別加入5個(gè)新鮮日本血吸蟲(chóng)蟲(chóng)卵、50μl日本血吸蟲(chóng)蟲(chóng)卵排泄分泌產(chǎn)物(egg secretion product,ESP)制備人工模擬陽(yáng)性糞樣。用糞DNA提取試劑盒分別提取加入蟲(chóng)卵的未經(jīng)研磨或研磨2 min后的人工模擬陽(yáng)性糞樣、加入蟲(chóng)卵ESP的糞樣和陰性糞樣的DNA,以日本血吸蟲(chóng)28S核糖體DNA(r DNA)為檢測(cè)靶基因,用LAMP法、PCR法檢測(cè),評(píng)估LAMP法檢測(cè)糞樣中日本血吸蟲(chóng)蟲(chóng)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(chóng)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(cè),并與PCR法、孵化法的檢測(cè)結(jié)果進(jìn)行比較,評(píng)價(jià)其檢測(cè)流行區(qū)現(xiàn)場(chǎng)野外糞樣的效果。結(jié)果LAMP法檢測(cè)加入蟲(chóng)卵糞樣并經(jīng)研磨后提取的DNA、加入蟲(chóng)卵ESP糞樣提取的DNA均為陽(yáng)性反應(yīng),呈綠色;而加入蟲(chóng)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(cè)的結(jié)果與LAMP法檢測(cè)結(jié)果相近。LAMP法、PCR法和孵化法檢測(cè)血吸蟲(chóng)病流行區(qū)牛野外糞樣的陽(yáng)性率分別為5.43%(12/221)、4.52%(10/221)、0.90%(2/221)。LAMP法的陽(yáng)性率明顯高于孵化法(P0.05),LAMP法與PCR法陽(yáng)性率差異無(wú)統(tǒng)計(jì)學(xué)意義(P0.05)。結(jié)論 LAMP法可用于檢測(cè)流行區(qū)現(xiàn)場(chǎng)野外糞樣的日本血吸蟲(chóng)蟲(chóng)卵DNA,其現(xiàn)場(chǎng)應(yīng)用價(jià)值有待進(jìn)一步驗(yàn)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(guó)疾病預(yù)防控制中心寄生蟲(chóng)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(guó)家級(jí)熱帶病國(guó)際聯(lián)合研究中心衛(wèi)生部寄生蟲(chóng)病原與媒介生物學(xué)重點(diǎn)實(shí)驗(yàn)室;湖南省血吸蟲(chóng)病防治所;
【基金】：上海市衛(wèi)生和計(jì)劃生育委員會(huì)科研課題(No.201440498) 湖南省科技廳項(xiàng)目(No.2012FJ4103) 國(guó)家衛(wèi)生計(jì)生委疾控局委托項(xiàng)目(2015-2016) 中國(guó)疾控中心寄生蟲(chóng)病所防治技術(shù)儲(chǔ)備科研基金(No.CB-15005)~~
【分類號(hào)】：R383.24

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