中国血吸虫病防治杂志
中國血吸蟲病防治雜誌
중국혈흡충병방치잡지
CHINESE JOURNAL OF SCHISTOSOMIASIS CONTROL
2014年
5期
522-525,530
,共5页
王飞%田茵%杨静%孙福军%孙宁%刘必永%田睿%葛广路%邹明强%邓丛良%刘翌
王飛%田茵%楊靜%孫福軍%孫寧%劉必永%田睿%葛廣路%鄒明彊%鄧叢良%劉翌
왕비%전인%양정%손복군%손저%류필영%전예%갈엄로%추명강%산총량%류익
恶性疟原虫%输入性%纳米磁分离%实时荧光定量PCR%检测
噁性瘧原蟲%輸入性%納米磁分離%實時熒光定量PCR%檢測
악성학원충%수입성%납미자분리%실시형광정량PCR%검측
Plasmodium falciparum%Imported%Magnetic nanoparticles separation%Real-time PCR%Detection
目的：建立恶性疟原虫纳米磁分离实时荧光定量PCR检测方法，快速准确地检测恶性疟原虫，为输入性恶性疟防控提供技术支持。方法根据恶性疟原虫基因组18S rRNA保守区序列，设计并合成引物和探针；构建质粒标准品，拟合标准曲线，采用纳米磁分离法提取核酸，建立恶性疟原虫纳米磁分离实时荧光定量PCR检测法，并进行该方法的灵敏度和特异性评价。结果与常规法（镜检法和快检法）相比，该方法检测恶性疟原虫更加灵敏，并具有良好的特异性，在（2.5×101~2.9×108）copies/ml拷贝数范围内循环阈值（Ct值）与质粒标准品拷贝数的对数值之间存在良好的线性关系（R2=0.999）；应用该方法从非洲维和部队13名归国人员中检出1例低水平恶性疟原虫感染者，而常规法（镜检法和快检法）未检出。结论该方法能够快速准确地检测恶性疟原虫，在口岸低密度恶性疟原虫感染者的检测中具有很大的应用价值。
目的：建立噁性瘧原蟲納米磁分離實時熒光定量PCR檢測方法，快速準確地檢測噁性瘧原蟲，為輸入性噁性瘧防控提供技術支持。方法根據噁性瘧原蟲基因組18S rRNA保守區序列，設計併閤成引物和探針；構建質粒標準品，擬閤標準麯線，採用納米磁分離法提取覈痠，建立噁性瘧原蟲納米磁分離實時熒光定量PCR檢測法，併進行該方法的靈敏度和特異性評價。結果與常規法（鏡檢法和快檢法）相比，該方法檢測噁性瘧原蟲更加靈敏，併具有良好的特異性，在（2.5×101~2.9×108）copies/ml拷貝數範圍內循環閾值（Ct值）與質粒標準品拷貝數的對數值之間存在良好的線性關繫（R2=0.999）；應用該方法從非洲維和部隊13名歸國人員中檢齣1例低水平噁性瘧原蟲感染者，而常規法（鏡檢法和快檢法）未檢齣。結論該方法能夠快速準確地檢測噁性瘧原蟲，在口岸低密度噁性瘧原蟲感染者的檢測中具有很大的應用價值。
목적：건립악성학원충납미자분리실시형광정량PCR검측방법，쾌속준학지검측악성학원충，위수입성악성학방공제공기술지지。방법근거악성학원충기인조18S rRNA보수구서렬，설계병합성인물화탐침；구건질립표준품，의합표준곡선，채용납미자분리법제취핵산，건립악성학원충납미자분리실시형광정량PCR검측법，병진행해방법적령민도화특이성평개。결과여상규법（경검법화쾌검법）상비，해방법검측악성학원충경가령민，병구유량호적특이성，재（2.5×101~2.9×108）copies/ml고패수범위내순배역치（Ct치）여질립표준품고패수적대수치지간존재량호적선성관계（R2=0.999）；응용해방법종비주유화부대13명귀국인원중검출1례저수평악성학원충감염자，이상규법（경검법화쾌검법）미검출。결론해방법능구쾌속준학지검측악성학원충，재구안저밀도악성학원충감염자적검측중구유흔대적응용개치。
Objective To establish a magnetic nanoparticles separation-based quantitative real-time PCR(RT-PCR)assay for fast and accurate detection of Plasmodium falciparum and providing a technical support for improving the control and preven-tion of imported malaria. Methods According to the conserved sequences of the P. falciparum genome 18SrRNA,the species-specific primers and probe were designed and synthetized. The RT-PCR was established by constructing the plasmid standard , fitting the standard curve and using magnetic nanoparticles separation. The sensitivity and specificity of the assay were evaluat-ed. Results The relationship between the threshold cycle(Ct)and logarithm of initial templates copies was linear over a range of 2.5×101 to 2.5×108 copies/μl(R2=0.999). Among 13 subjects of entry frontier,a P. falciparum carrier with low load was de-tected by using the assay and none was detected with the conventional examinations(microscopic examinations and rapid tests). Conclusion This assay shows a high sensitivity in detection of P. falciparum,with rapid and accurate characteristics,and is especially useful in diagnosis of P. falciparum infectors with low parasitaemia at entry-exit frontier ports.
