漢坦病毒基因分型和應用重組抗原進行HFRS抗體快速檢測的研究
發(fā)布時間:2019-01-03 08:14
【摘要】:本研究使用RT-nested PCR分型檢測方法及核苷酸序列測定技術,對來源于福建省內(nèi)HFRS監(jiān)測點的鼠肺標本及病人血清進行基因分型并對部分標本的核苷酸序列進行分析比較。RT-PCR分型檢測結果顯示:24份陽性鼠肺標本中檢出率為95.8%;20份病人血清標本中僅11份擴增陽性,檢出率分別為:83%(≤1周),12.5%(1周)。擴增陽性標本中僅1份RT-PCR分型為HTN型,其余均為SEO型,這與核苷酸序列分型結果相一致。表明近年福建省流行的漢坦病毒仍以SEO型為主。核酸序列比較分析發(fā)現(xiàn),福建省內(nèi)同一地區(qū)流行的SEO型漢坦病毒核苷酸的同源性很高,大于99%,而不同地區(qū)病毒間核酸序列變異比較大,尤其是永春和松溪這兩個地區(qū)的毒株差異達到20%左右,可能是SEO型中兩個新的亞型。 在E.coli中表達漢坦病毒A_(537)株NP重組抗原,采用電洗脫、親和層析等方法純化重組蛋白,純化的重組蛋白e112直接包被聚乙烯96孔板,用于間接法ELISA檢測HFRS患者血清中IgG抗體,與IFAT比較,其敏感性和特異性分別為94.7%、95.6%,適用于大規(guī)模的流行病學調查;或以膠體金標記重組抗原,運用金標快速免疫層析法可同時檢測HFRS患者血清中IgM和IgG抗體,與IFAT/ELISA比較,其敏感性和特異性分別為97.9%、98.2%,具有簡便、快速等優(yōu)點,適用于各種層次尤其是基層醫(yī)療單位、農(nóng)村衛(wèi)生院等缺乏實驗條件和專業(yè)人員的地方對HFRS疑似患者作出早期診斷。
[Abstract]:In this study, RT-nested PCR typing and nucleotide sequencing were used. The genotyping of rat lung and patient serum from HFRS monitoring sites in Fujian province was carried out and the nucleotide sequences of some samples were analyzed and compared. The results of RT-PCR typing showed that the detection rate of 24 positive rat lung samples was 95.8%. The positive rate of amplification was 83% (鈮,
本文編號:2399107
[Abstract]:In this study, RT-nested PCR typing and nucleotide sequencing were used. The genotyping of rat lung and patient serum from HFRS monitoring sites in Fujian province was carried out and the nucleotide sequences of some samples were analyzed and compared. The results of RT-PCR typing showed that the detection rate of 24 positive rat lung samples was 95.8%. The positive rate of amplification was 83% (鈮,
本文編號:2399107
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