1.人兽共患寄生虫

1.1A Fluorescent Recombinase Aided AmplificationAssay for Detection of Babesia microt

1.2Rapid Visual Detection of Plasmodium Using Recombinase-Aided Amplification With Lateral Flow DipstickAssay

1.3重组酶介导恒温扩增技术检测疟原虫方法的建立

1.4实时荧光重组酶介导核酸扩增在疟原虫快速检测中的应用研究

1.5重组酶介导的斯氏并殖吸虫等温扩增荧光检测方法的建立及检测效果初步评价

1.6重组酶介导的日本血吸虫特异性基因片段核酸等温扩增检测方法的建立

1.7结合重组酶介导的核酸等温扩增和荧光探针快速检测日本血吸虫基因片段

1.8加强分子诊断方法的研发应用助力我国精准血防

1.9重组酶介导的核酸等温扩增荧光法快速检测日本血吸虫感染性钉螺

1.10重组酶介导的隐孢子虫属特异性等温核酸扩增方法的建立及评价

1.11重组酶介导的华支睾吸虫特异性核酸等温扩增方法的建立及初步评价

1.12再送“瘟神”:消除血吸虫病适宜技术最新研究进展

1.13重组酶介导的等温核酸扩增技术检测多房棘球绦虫方法的建立及初步应用

1.14重组酶介导等温核酸扩增技术检测细棘球绦虫方法的建立及初步应用评价

1.15基于重组酶介导等温扩增技术的广州管圆线虫核酸检测方法的建立

1.16基于重组酶介导等温扩增技术的细粒棘球绦虫核酸检测方法的建立

1.17基于重组酶介导核酸等温扩增反应的曼氏血吸虫基因检测方法的建立

1.18新型等温扩增技术推动寄生虫病现场快速检测能力提升

1.19重组酶介导的蓝氏贾第鞭毛虫特异性等温核酸扩增方法的建立及评价

1.20基于荧光重组酶介导等温扩增技术的利什曼原虫核酸检测方法的建立

1.21基于重组酶介导核酸等温扩增技术的日本血吸虫特异性基因片段核酸试纸条检测方法的建立

1.22快速检测田鼠巴贝西虫重组酶介导核酸等温扩增方法的建立

1.23 重组酶介导的核酸等温扩增荧光法检测日本血吸虫感染性钉螺的效能评价

1.24重组酶介导等温扩增技术检测疟原虫方法的建立和评价

1.25重组酶介导核酸等温扩增荧光法用于日本血吸虫感染性钉螺旱期检测的研究

A Fluorescent Recombinase Aided Amplifica-tion Assay for Detection of Babesia microti

Abstract Babesia microti is one of the most common causative agents of babesiosis.A sensitive and raplddetection is necessary for screening potentially infected individuals.In this study,B. microti cytochrome coxldase subunit l(cox1) was selected as the target gene,multiple primers were designed,and optimized by arecombinase-aided amplification(RAA) assay.The optimal primers and probe were labeled with fluorescein.The sensitivity of fluorescent RAAGRAA) was evaluated using gradient diluents of the cox1 recombinantplasmid and genomic DNA extracted from whole blood of B.microti infected mice.The spedificity offRAA wasassessed by other transfusion transmitted parasites.The analytical sensitivity of the fRAA assay was 10 coplesof recombinant plasmid per reaction and 10 fg/ul B.microti genomic DNA.No cross-re action with any otherblood-transmitted parasites was observed.Our results demonstrated that the fRAA assay would be rapid,sensitive,and specific for the detection of B.microti.

Key words:Babesia microti,recombinase-aided amplification,molecular detection.

Rapid Visual Detection ofPlasmodium Using Recombinase Aided Am-plification With Lateral Flow Dipstick Assay

Background:Malaria is a global public health problem.China has had no case of indigenous malaria since 2016.Hawever,imported cases of malaria remain an issue amang travelers,overseas workers,and foreign traders.Although these cases are always asymptomatic,if they donate blood,there is a great risk of transfusion trans-mitted-malaria(TTM).Therefore,blood banks need a rapid screening tool to detect Pasmodium species.Methods:We designed an assay using recombinase-aided amplification(RAA) and a lateral-flow dipstick(LFD)(RAA-LFD) to detect the 18S ribasomal RNA gene of Plasmodium species.Sensitivity was evaluated using arecombinant plasmid and Plasmodium genomic DNA.Specificity was evaluated using DNA extracted from theblood of patients with malaria ar other infectious parasites.For clinical assessment,blood samples frompatients with malaria and blood donors were evalu ated.Results:The RAA-LFD assay was performed in an incubator block at 37C for 15 min,and the amplicans werevisible to the naked eye an the flaw dipsticks within 3 min.The sensitivity was 1 copy/mL of recombinantplasmid.For genomic DNA from whole blood of malaria patients infected with P.falciparum, P.vivax,P. ovale,and P.malariae,the sensitivity was0.1 pg/mL,10 pg/mL,10-100 pg/mL,and 100pg/mL,respectively.Thesensi-tivity of this assay was 100pg/mL.No cross-reaction withother transfusion transmissible parasites wasdetect-ed.Conclusions:The results demonstrated that this RAA-LFD assay was suitable for reliable field detection ofPlasmodium spedes in low-resource settings with limited laboratory capabilities.

Keywords:malaria,nucleic add detection,plasmodium,recombinase-aided amplification, lateralflow dipstick.

重组酶介导恒温扩增技术检测疟原虫方法的建立

摘要：本研究利用重组酶介导恒温扩增RAA技术，选取疟原虫18SrDNA序列，设计了检测疟原虫的通用引物，并对引物进行筛选和对其特异性进行检测。筛选出4组扩增效果较好的引物。该方法整个反应过程在37℃下进行，扩增时间短(40min)，并具有良好的特异性。成功建立了一种检测疟原虫的RAA法，并且该方法适合于进出口岸疟原虫的快速检测。

关键词：重组酶介导扩增；疟原虫；分子检测；疟疾。

重组酶介导的斯氏并殖吸虫等温扩增荧光检测方法的建立及检测效果初步评价

摘要：目的建立一种基于重组酶介导的等温扩增(recombinase-aided isothermal amplific ation,RAA)技术的斯氏并殖吸虫快速核酸检测方法，并对其检测效果进行初步评价。方法从溪蟹样本中分离出斯氏并殖吸虫、卫氏并殖吸虫和三平正并殖吸虫囊坳,提取基因组DNA进行分子鉴定。以斯氏并殖吸虫线粒体细胞色素c氧化酶亚基I基因(cytochrome coxidase 1, cox1)基因序列作为靶序列设计、制备、筛选引物及探针。以河南省济源市和洛阳市宜阳县斯氏并殖吸虫囊蜕基因组DNA为模板进行荧光RAA检测方法验证。以含斯氏并殖吸虫cox1基因序列的不同浓度重组质粒和斯氏并殖吸虫囊坳基因组DNA为模板进行荧光RAA扩增，评价其检测灵敏度；应用建立的荧光RAA法同时检测卫氏并殖吸虫、三平正并殖吸虫、华支睾吸虫和日本血吸虫基因组DNA，评价其检测特异性。结果从溪蟹样本中分离出斯氏并殖吸虫、卫氏并殖吸虫和三平正并殖吸虫囊蝴，经分子鉴定和系统进化分析确认其与GenBank中并殖吸虫标准株基因序列具有同源性。成功建立了斯氏并殖吸虫荧光RAA检测方法，可在5min内扩增到河南省济源市和洛阳市宜阳县采集的斯氏并殖吸虫囊鲋基因组DNA，而阴性对照无扩增。以重组质粒为模板,荧光RAA法最低检出限为10拷贝/L重组质粒,且均在5 min内出现阳性扩增；以基因组DNA为模板,可检测到的最低模板DNA浓度为10 pgluL，且均在10~15min内出现阳性扩增。荧光RAA法检测卫氏并殖吸虫、三平正并殖吸虫、日本血吸虫和华支睾吸虫基因组DNA结果均为阴性。结论成功建立了一种基于荧光RAA技术的快速、敏感、特异的斯氏并殖吸虫核酸检测方法,在斯氏并殖吸虫病流行区溪蟹现场快速检测与虫种鉴定中具有潜在应用价值。

关键词：斯氏并殖吸虫；重组酶介导的等温扩增；核酸检测；检测效果。