本文選題：日本血吸蟲　切入點：TaqMan實時定量PCR　出處：《中國人獸共患病學(xué)報》2014年03期 　論文類型：期刊論文

【摘要】：目的以日本血吸蟲高度重復(fù)序列-逆轉(zhuǎn)錄轉(zhuǎn)座子SjR2為靶序列,建立TaqMan實時定量PCR法檢測宿主血清中的日本血吸蟲DNA用于評價感染度。方法以real-time PCR法檢測不同感染度的家兔模型血清DNA。結(jié)果該法具有高度的敏感性和特異性,可以檢測到44.7拷貝的重組質(zhì)粒DNA。在感染家兔后的3d到7周血清中均能檢測到血吸蟲DNA,不同感染度早期檢測的時間節(jié)點及其檢測閾值分別為,家兔感染30條尾蚴(EPG=14)需在感染后2周才能檢測到目的DNA,其含量為119.03拷貝,感染50條尾蚴(EPG=24)、感染100條尾蚴(EPG=48)則在感染后1周可檢測到目的DNA,其含量分別為54.36拷貝和72.24拷貝,在家兔感染200條尾蚴(EPG=97)、感染500條尾蚴(EPG=232)最早在感染后3d即可檢測到目的 DNA,其含量分別為60.34拷貝和142.47拷貝。日本血吸蟲感染宿主血清DNA水平與感染度呈正相關(guān),即血清DNA濃度隨感染度的增大而上升。結(jié)論本研究所建立的real-time PCR法可定量檢測血清DNA動態(tài)變化并對日本血吸蟲病診斷及感染度的評價具有潛在的應(yīng)用價值,為日本血吸蟲病診斷與療效考核提供了新方法。
[Abstract]:Objective to take the highly repetitive sequence of Schistosoma japonicum reverse transcription transposon SjR2 as the target sequence. A real-time quantitative TaqMan PCR method was established for the detection of schistosomiasis japonicum DNA in host serum. Methods real-time PCR assay was used to detect serum DNA in rabbits with different infection degrees. The results showed that the method was highly sensitive and specific to the infection of Schistosoma japonicum. The DNA of Schistosoma japonicum could be detected in the sera of rabbits from 3 days to 7 weeks after infection. The time node of early detection of different infection degree and the detection threshold were respectively. In rabbits infected with 30 cercariae, the target DNA was detected only 2 weeks after infection. The content of the target DNA was 119.03 copies, 50 cercariae were infected with EPGM24, 100 cercariae were infected with EPG4), and the target DNA was detected 1 week after infection, the contents of which were 54.36 copies and 72.24 copies, respectively. In rabbits infected with 200 cercariae and 500 cercariae, the target DNA was detected at the earliest 3 days after infection, the contents of which were 60.34 copies and 142.47 copies, respectively. The serum DNA level of Schistosoma japonicum infected host was positively correlated with the degree of infection. Conclusion the real-time PCR method established in this study can quantitatively detect the dynamic changes of serum DNA and has potential application value in the diagnosis and infection evaluation of schistosomiasis japonicum. It provides a new method for diagnosis and curative effect assessment of schistosomiasis japonicum.
【作者單位】： 蘇州大學(xué)醫(yī)學(xué)部基礎(chǔ)醫(yī)學(xué)與生物科學(xué)學(xué)院病原生物學(xué)系;
【基金】：國家973計劃項目子課題(No.2007CB513102)~~
【分類號】：R346

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相關(guān)期刊論文 前3條

1 劉愛平;楊巧林;郭俊杰;許靜;夏超明;;巢式PCR法檢測日本血吸蟲低感染度宿主血清DNA的研究[J];蘇州大學(xué)學(xué)報(醫(yī)學(xué)版);2010年05期

2 陳一平;翁心華;徐肇s，

本文編號：1623153