小反刍动物(PPRV),一种高度传染性的病毒性疾病,对绵羊和山羊造成重大经济损失。最近,PPR病毒(PPRV),采用了新的宿主,PPRV的谱系IV代表了同一谱系内的遗传多样性。350个样品,包括血,拭子,和绵羊/山羊的组织,是在巴基斯坦2020-2021年疾病暴发期间收集的。这些样品通过RT-PCR和三个具有登录号的PPRV分离物进行分析,MW600920、MW600921和MW600922已提交给GenBank,基于部分N基因测序。该分析提供了对遗传特征的更好理解和用于快速PPRV诊断的靶向RT-PCR方法。使用在Vero细胞中生长的半纯化抗原MW600922分离物开发IELISA测试。PPRV分离株目前与土耳其菌株存在高度分歧;相反,从巴基斯坦收集的分离株的相似性相当于99.73%。开发的间接ELISA(IELISA)测试表明,抗体(血清)稀释度为1:200,抗原稀释度为1:32时的抗体检测率。与cELISA相比,观察到高特异性(85.23%)和敏感性(90.60%)。与病毒中和试验(VNT)相反,观察到IELISA在其结果中具有100%的特异性和82.14%的敏感性。基于这些结果,使用IELISA对PPR抗体进行血清学调查可能是更大规模的更有效策略.此外,我们的研究结果表明,在成本效益和存储效率方面的研究取得了重大突破,开发的IELISA测试强烈建议在发展中国家使用。
小反刍动物(PPRV)是一种跨界动物,高度传染性,以及影响小反刍动物和野生动物的经济上重要的病毒性疾病。PPRV,一种只针对动物的疾病,是全球根除计划(PPRVGEP)的重点,其目标是到2030年根除这种疾病。GEP第一阶段(2017-2021年)完成后,巴基斯坦已启动第二阶段:PPRV的存在和控制策略的实施。快速准确的实验室诊断对疾病的有效控制和根除至关重要。在本研究中,我们通过逆转录聚合酶链反应(RT-PCR)改善了诊断,这不仅可以检测低病毒浓度,而且有助于谱系IV病毒的遗传分析。然而,开发具有成本效益的间接ELISA(iELISA)可以分析从更大的小反刍动物群体中获得的血清样本。
Peste des petits ruminants (PPRV), a highly contagious viral disease, causes significant economic losses concerning sheep and goats. Recently, PPR viruses (PPRVs), have adopted new hosts and lineage IV of PPRVs represents genetic diversity within the same lineage. 350 samples, including blood, swabs, and tissues from sheep/goats, were collected during the 2020-2021 disease outbreaks in Pakistan. These samples were analysed through RT-PCR and three isolates of PPRV with accession numbers, MW600920, MW600921, and MW600922, were submitted to GenBank, based on the partial N-gene sequencing. This analysis provides a better understanding of genetic characterizations and a targeted RT-PCR approach for rapid PPRV diagnosis. An
IELISA test was developed using the semi-purified antigen MW600922 isolate grown in Vero cells. The PPRV isolates currently present high divergence with the Turkish strain; conversely, similarities equivalent to 99.73% were observed for isolates collected from Pakistan. The developed indirect ELISA (
IELISA) test demonstrated antibody detection rates at dilutions of 1:200 for antibodies (serum) and 1:32 for antigens. In comparison to cELISA, high specificity (85.23%) and sensitivity (90.60%) rates were observed. In contrast to the virus neutralization test (VNT),
IELISA was observed to be 100% specific and 82.14% sensitive in its results. Based on these results, serological surveys conducted for PPR antibodies using IELISA can be a more effective strategy on a larger scale. Furthermore, our results demonstrate a significant breakthrough in the research in terms of cost-effectiveness and storage efficiency, and the developed
IELISA test is highly recommended for use in developing countries.
Peste des petits ruminants (PPRV) is a transboundary, highly contagious, and economically significant viral disease affecting small ruminants and wildlife. PPRV, a disease that only targets animals, is the focus of the Global Eradication Programme (PPRV GEP), which aims to eradicate the disease by 2030. Following the completion of the first phase of the GEP (2017–2021), Pakistan has initiated the second phase: PPRV presence and the implementation of a control strategy. Rapid and accurate laboratory diagnosis is vital to the disease’s effective control and eradication. In the present study, we have improved diagnosis by reverse transcriptase polymerase chain reaction (RT-PCR), which not only can detect low viral concentrations but also contributes to the genetic analysis of lineage-IV viruses. However, the development of cost-effective indirect ELISA (
iELISA) may allow for the analysis of serum samples obtained from larger populations of small ruminants.