漢坦病毒基因分型和應(yīng)用重組抗原進(jìn)行HFRS抗體快速檢測的研究
發(fā)布時(shí)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(shù),對(duì)來源于福建省內(nèi)HFRS監(jiān)測點(diǎn)的鼠肺標(biāo)本及病人血清進(jìn)行基因分型并對(duì)部分標(biāo)本的核苷酸序列進(jìn)行分析比較。RT-PCR分型檢測結(jié)果顯示:24份陽性鼠肺標(biāo)本中檢出率為95.8%;20份病人血清標(biāo)本中僅11份擴(kuò)增陽性,檢出率分別為:83%(≤1周),12.5%(1周)。擴(kuò)增陽性標(biāo)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(jié)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(gè)地區(qū)的毒株差異達(dá)到20%左右,可能是SEO型中兩個(gè)新的亞型。 在E.coli中表達(dá)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(xué)調(diào)查;或以膠體金標(biāo)記重組抗原,運(yùn)用金標(biāo)快速免疫層析法可同時(shí)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(diǎn),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(shí)驗(yàn)條件和專業(yè)人員的地方對(duì)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(hào):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(hào):2399107
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