發(fā)布時(shí)間：2018-06-20 05:15

  本文選題：腺病毒科 + 重組酶聚合酶擴(kuò)增技術(shù)�。� 參考：《軍事醫(yī)學(xué)》2017年07期

【摘要】：目的建立一種快速、準(zhǔn)確、可視化、易開展的B、E型腺病毒檢測(cè)技術(shù)。方法針對(duì)B、E型腺病毒殼蛋白保守序列設(shè)計(jì)通用引物,建立可同時(shí)檢測(cè)出這兩種腺病毒型別的重組酶聚合酶擴(kuò)增結(jié)合側(cè)流層析試紙條(recombinase polymerase amplification combined with a lateral flow dipstick,LFD-RPA)檢測(cè)技術(shù),評(píng)價(jià)其靈敏度與特異性,并驗(yàn)證最佳反應(yīng)溫度與反應(yīng)時(shí)間,同時(shí)利用該法對(duì)19例B、E型腺病毒感染患者及10例健康志愿者鼻咽拭子標(biāo)本進(jìn)行檢測(cè)。結(jié)果腺病毒LFD-RPA法檢測(cè)靈敏度可達(dá)10拷貝/μl,與q PCR方法相近,且不會(huì)與其他亞屬腺病毒及其他病毒發(fā)生交叉反應(yīng)。反應(yīng)可在25~45℃溫度范圍內(nèi)高效進(jìn)行,檢測(cè)全程僅需15~20 min。利用臨床咽拭子標(biāo)本對(duì)檢測(cè)體系進(jìn)行評(píng)估,其靈敏度及特異性均達(dá)100%。結(jié)論該檢測(cè)方法靈敏度高、特異性強(qiáng)、操作簡(jiǎn)單,反應(yīng)快速,能脫離對(duì)大型儀器、專業(yè)操作人員及正規(guī)實(shí)驗(yàn)室的依賴,在基層衛(wèi)生部門,尤其在野外及現(xiàn)場(chǎng)檢測(cè)中具有極大的應(yīng)用潛力。
[Abstract]:Objective to establish a rapid, accurate, visual and easy detection technique for adenovirus type E (AV). Methods A universal primer was designed for the conserved sequence of adenovirus type E adenovirus shell protein, and a recombinant enzyme polymerase amplification method combined with LFD-RPAA assay was developed to detect the two adenovirus types simultaneously. The recombinant polymerase amplification combined with side flow chromatography test strip was used to detect the recombinant adenovirus types. The sensitivity and specificity were evaluated, and the optimum reaction temperature and reaction time were verified. At the same time, the nasopharyngeal swabs of 19 patients with adenovirus type E infection and 10 healthy volunteers were detected by this method. Results the sensitivity of LFD-RPA assay was up to 10 copies / 渭 l, which was similar to that of Q PCR, and no cross reaction with other subgenus adenovirus and other viruses was observed. The reaction can be carried out efficiently in the temperature range of 25 鈩，

本文編號(hào)：2043106