【摘要】：為建立快速檢測(cè)駑巴貝斯蟲Bc-48基因的方法,本研究根據(jù)Bc-48保守基因序列設(shè)計(jì)合成特異性引物和MGB熒光探針,以構(gòu)建的重組質(zhì)粒作為陽性標(biāo)準(zhǔn)品,經(jīng)條件優(yōu)化后建立了檢測(cè)Bc-48基因的TaqMan-MGB熒光定量PCR方法。結(jié)果顯示,該方法能夠特異性檢測(cè)駑巴貝斯蟲,而對(duì)其它病原檢測(cè)均為陰性;該方法最小檢出量為5.17×10~1拷貝/μL的標(biāo)準(zhǔn)質(zhì)粒,敏感性是常規(guī)PCR的100倍;組內(nèi)與組間重復(fù)性試驗(yàn)的變異系數(shù)均小于1%,重復(fù)性好;采用所建立的方法和常規(guī)PCR分別對(duì)90份臨床樣品進(jìn)行檢測(cè),結(jié)果顯示本研究建立的駑巴貝斯蟲TaqMan-MGB熒光定量PCR檢測(cè)方法特異性強(qiáng)、敏感性高、重復(fù)性好,可以用于駑巴貝斯蟲病臨床樣品診斷、流行病學(xué)調(diào)查。本研究為蜱傳馬梨形蟲病的防控提供新的方法。
[Abstract]:In order to establish a rapid method for detecting the Bc-48 gene of Babesia caballi, specific primers and MGB fluorescence probes were designed and synthesized according to the conserved gene sequence of Bc-48. The recombinant plasmid was used as the positive standard. A TaqMan-MGB fluorescence quantitative PCR method for the detection of Bc-48 gene was established after optimization of the conditions. The results showed that the method could detect Babesia caballi specifically, but negative for other pathogens, and the sensitivity of the standard plasmid with a minimum detection quantity of 5.17 脳 10 渭 1 copy / 渭 L was 100 times higher than that of conventional PCR. The coefficient of variation of intra-group and inter-group repeatability test was less than 1%, and the repeatability was good. The established method and routine PCR were used to detect 90 clinical samples respectively. The results showed that the method established in this study was highly specific, sensitive and reproducible for the detection of Babesia caballi TaqMan-MGB. It can be used for the diagnosis and epidemiological investigation of Babes caballi disease. This study provides a new method for the prevention and control of tick-borne pear disease.
【作者單位】： 新疆農(nóng)業(yè)大學(xué)動(dòng)物醫(yī)學(xué)學(xué)院;伊犁出入境檢驗(yàn)檢疫局綜合技術(shù)服務(wù)中心;
【基金】：國(guó)家自然基金NSFC-新疆聯(lián)合基金重點(diǎn)項(xiàng)目(U1403283)
【分類號(hào)】：S852.7

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