【摘要】：目的了解小鼠腦脊髓炎病毒(TMEV)自然感染情況,探究人工感染TMEV小鼠體內(nèi)各臟器組織中病毒分布及血清抗體變化。方法采用酶聯(lián)免疫吸附試驗(yàn)(ELISA)和熒光定量RT-PCR(qRT-PCR)檢測方法對2010年~2015年廣東地區(qū)采集的SPF級小鼠、開放環(huán)境飼養(yǎng)的小鼠以及野生褐家鼠臨床樣本進(jìn)行TMEV檢測。36只ICR小鼠經(jīng)腦內(nèi)接種TMEV BeAn病毒,每天觀察動物的臨床癥狀,在接種第0、3、7、10、17、21、31、39、46天每個時間點(diǎn)分別對3只動物安樂死,剖檢并取血清和組織臟器樣本進(jìn)行TMEV檢測。結(jié)果 SPF級小鼠TMEV抗體陽性率為5.29%(n=2834),核酸陽性率為27.27%(n=457);開放環(huán)境飼養(yǎng)的小鼠的抗體和核酸陽性率分別為71.95%(n=82)和53.66%(n=82);野生褐家鼠中核酸陽性率為25.93%(n=27)。TMEV陽性小鼠中僅有兩只小鼠表現(xiàn)有明顯的臨床癥狀。盲腸內(nèi)容物、糞便和腦是qRT-PCR檢測的最佳選擇樣本。ICR小鼠腦內(nèi)接種TMEV BeAn病毒后第3 d可在腦、心臟、肝臟、肺臟和胃中檢測到病毒核酸,脾臟、腎臟和盲腸中未檢測到病毒核酸。肝臟、心臟、肺臟和胃中的病毒在接種后第10天已完全清除,腦中的病毒一直持續(xù)存在到第46天試驗(yàn)結(jié)束。小鼠感染后第7天可以檢測到抗體,隨后抗體水平逐漸升高,接種后17 d抗體陽性率達(dá)100%,并一直到46 d都可以維持較高的抗體水平。人工感染小鼠呈隱性感染,臨床上并未表現(xiàn)明顯癥狀和眼觀病理變化。結(jié)論廣東地區(qū)實(shí)驗(yàn)小鼠和野生褐家鼠均存在TMEV感染,且感染率較高。小鼠接種TMEV BeAn毒株后呈隱性感染,感染小鼠第7天可以產(chǎn)生抗體且持續(xù)存在。病毒在感染小鼠肝臟、心臟、肺臟和胃中短時間存在,而在腦中長期存在。qRT-PCR與ELISA兩種檢測方法具有較好的一致性,qRT-PCR檢測方法可作為實(shí)驗(yàn)動物國家標(biāo)準(zhǔn)的有力補(bǔ)充。
[Abstract]:Objective to investigate the natural infection of encephalomyelitis virus (TMEV) in mice and to explore the distribution of virus and the change of serum antibody in organs and tissues of mice artificially infected with TMEV. Methods enzyme-linked immunosorbent assay (ELISA) and fluorescence quantitative RT-PCR (qRT-PCR) were used to detect TMEV in clinical samples of SPF mice, mice raised in open environment and wild brown mice collected from Guangdong from 2010 to 2015. 36 ICR mice were inoculated with TMEV BeAn virus in the brain, and the clinical symptoms of the animals were observed every day. 3 animals were euthanized at 0, 3, 7, 10, 17, 21, 31, 39, 46 days, respectively. Serum and tissue and organ samples were taken for TMEV detection. Results the positive rate of TMEV antibody was 5.29% (n 鈮，

本文編號：2500191