Development and evaluation of recombinase polymerase amplification combined with lateral flow dipstick assays for co-detection of epizootic haemorrhagic disease virus and the Palyam serogroup virus.
Development and evaluation of recombinase polymerase amplification combined with lateral flow dipstick assays for co-detection of epizootic haemorrhagic disease virus and the Palyam serogroup virus.
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重组酶聚合酶扩增与侧流试纸检测联合检测流行性出血病病毒和 Palyam 血清群病毒的开发和评估
DOI：
10.1186/s12917-021-02977-9
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发表时间：
2021-08-25
影响因子：
2.6
通讯作者：
Liao DF
中科院分区：
文献类型：
--
作者：
Li ZR;Yang ZX;Li ZH;Gao X;Hu ZY;Yang H;Liao DF
背景流行性出血症病毒（EHDV）和Palyam血清群病毒（PALV）在全球范围内造成了与畜牧业生产相关的重大经济损失。一个快速，灵敏和特异的方法检测EHDV和PALV是病毒检测，监测，并成功控制和消除相关diseases.ResultsIn本研究中，重组酶聚合酶扩增结合侧流试纸条（RPA-LFD）检测EHDV和PALV的基因组片段1（Seg-1）的共同检测是至关重要的。建立的RPA-LFD检测EHDV和PALV的分析灵敏度分别为7.1拷贝/微升和6.8拷贝/微升。与非洲马瘟病毒、蓝舌病病毒、广西环状病毒、西藏环状病毒、云南环状病毒等环状病毒属成员无交叉反应。建立的RPA-LFD检测方法可准确检测5个血清型的39株EHDV和3个血清型的29株PALV。实时定量聚合酶链反应（qRT-PCR）与建立的RPA-LFD方法在哨牛上的溯源结果一致。对56份分离到EHDV或PALV的血液样品和96份从牛场采集的血液样品进行qRT-PCR检测，与建立的RPA-LFD检测方法的符合率均在94.8%以上。结果表明，建立的RPR-LFD检测方法特异、灵敏、可靠，可用于EHDV和PALV的早期临床诊断。该方法可作为EHDV和PALV现场诊断的一种快速、可靠、灵敏、低成本的方法。
BackgroundEpizootic haemorrhagic disease virus (EHDV) and the Palyam serogroup viruses (PALV) have led to significant economic losses associated with livestock production globally. A rapid, sensitive and specific method for the detection of EHDV and PALV is critical for virus detection, monitoring, and successful control and elimination of related diseases.ResultsIn the present study, a recombinase polymerase amplification combined with lateral flow dipstick (RPA-LFD) assay for the co-detection of genome segment 1 (Seg-1) of EHDV and PALV was developed and evaluated. The analytical sensitivities of the established RPA-LFD assay in the detection of EHDV and PALV were 7.1 copies/µL and 6.8 copies/µL, respectively. No cross-reaction with other members of the genusOrbivirus, including African horse sickness virus, bluetongue virus,Guangxi orbivirus, Tibet orbivirusandYunnan orbiviruswas observed. The established RPA-LFD assay accurately detected 39 EHDV strains belonging to 5 serotypes and 29 PALV strains belonging to 3 serotypes. The trace back results of quantitative real-time polymerase chain reaction (qRT-PCR) and the established RPA-LFD assay on sentinel cattle were consistent. The coincidence rates of qRT-PCR and the established RPA-LFD assay in 56 blood samples from which EHDV or PALV had been isolated and 96 blood samples collected from cattle farms were more than 94.8 %. The results demonstrated that the established RPR-LFD assay is specific, sensitive and reliable, and could be applied in early clinical diagnosis of EHDV and PALV.ConclusionsThis study highlights the development and application of the RPA-LFD assay in the co-detection of EHDV and PALV for the first time. The assay could be used as a potential optional rapid, reliable, sensitive and low-cost method for field diagnosis of EHDV and PALV.
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影响因子：
4.3
作者：
Fowler VL;Howson ELA;Flannery J;Romito M;Lubisi A;Agüero M;Mertens P;Batten CA;Warren HR;Castillo-Olivares J
通讯作者：
Castillo-Olivares J
开发重组酶聚合酶扩增结合侧流试纸法检测牛短暂热病毒
影响因子：
3.3
作者：
Hou P;Zhao G;Wang H;He C;Huan Y;He H
通讯作者：
He H
影响因子：
5
作者：
Bonneau, KR;Zhang, NZ;MacLachlan, NJ
通讯作者：
MacLachlan, NJ
影响因子：
3.8
作者：
Yamakawa, M;Kubo, M;Furuuchi, S
通讯作者：
Furuuchi, S
影响因子：
2.2
作者：
Yadin, H.;Brenner, J.;Mertens, P. P. C.
通讯作者：
Mertens, P. P. C.