Development of a highly sensitive point-of-care test for African swine fever that combines EZ-Fast DNA extraction with LAMP detection: Evaluation using naturally infected swine whole blood samples from Vietnam.
Development of a highly sensitive point-of-care test for African swine fever that combines EZ-Fast DNA extraction with LAMP detection: Evaluation using naturally infected swine whole blood samples from Vietnam.
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DOI：
10.1002/vms3.1124
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发表时间：
2023-05
影响因子：
1.7
通讯作者：
Yamazaki, Wataru
中科院分区：
文献类型：
--
作者：
Ngan, Mai Thi;Thi My Le, Huynh;Ngoc Dang, Vu;Thi Bich Ngoc, Trinh;Phan, Le Van;Thi Hoa, Nguyen;Quang Lam, Truong;Thi Lan, Nguyen;Notsu, Kosuke;Sekiguchi, Satoshi;Yamazaki, Yasuko;Yamazaki, Wataru
关键词：
虽然早期发现和早期遏制是控制非洲猪瘟（ASF）大流行的关键，但缺乏实用的现场检测方法是实现这一壮举的主要障碍。描述ASF的快速和灵敏的床旁检测（POCT）的开发，以及使用猪全血样本进行田间环境评价。总共从越南猪场采集了89份猪全血样本，并使用粗DNA提取和LAMP（环介导等温扩增）扩增的组合进行了POCT。POCT能够在10分钟内以极低的成本和相对容易地从猪全血样品中提取粗DNA。整个POCT从DNA提取开始到最终判断最多需要50 min。与传统的真实的时间PCR检测相比，POCT显示检测灵敏度降低1个对数，但诊断灵敏度为100%（56/56），诊断特异性为100%（33/33）。POCT更快，更容易执行，不需要特殊设备。该POCT有望促进ASF入侵的早期诊断和遏制，并在这两个地区流行和根除。开发的POCT更快，更容易执行，不需要特殊设备。我们在猪全血样本中验证了一种灵敏的ASF POCT，该POCT有望促进ASF入侵的早期诊断和遏制，无论是在地方流行还是在根除地区。
While early detection and early containment are key to controlling the African swine fever (ASF) pandemic, the lack of practical testing methods for use in the field are a major barrier to achieving this feat. To describe the development of a rapid and sensitive point‐of‐care test (POCT) for ASF, and its evaluation using swine whole blood samples for field settings. In total, 89 swine whole blood samples were collected from Vietnamese swine farms and were performed the POCT using a combination of crude DNA extraction and LAMP (loop‐mediated isothermal amplification) amplification. The POCT enabled crude DNA to be extracted from swine whole blood samples within 10 min at extremely low cost and with relative ease. The entire POCT required a maximum of 50 min from the beginning of DNA extraction to final judgment. Compared to a conventional real‐time PCR detection, the POCT showed a 1 log reduction in detection sensitivity, but comparable diagnostic sensitivity of 100% (56/56) and diagnostic specificity of 100% (33/33). The POCT was quicker and easier to perform and did not require special equipment. This POCT is expected to facilitate early diagnosis and containment of ASF invasion into both regions in which it is endemic and eradicated. The developed POCT is quicker and easier to perform and does not require special equipment. We validated a sensitive POCT for ASF in swine whole blood samples for field settings.This POCT is expected to facilitate early diagnosis and containment of ASF invasion into both regions in which it is endemic and eradicated.
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DOI：
10.3390/v13091717
发表时间：
2021-08-30
期刊：
Viruses
影响因子：
--
作者：
Sauter-Louis C;Conraths FJ;Probst C;Blohm U;Schulz K;Sehl J;Fischer M;Forth JH;Zani L;Depner K;Mettenleiter TC;Beer M;Blome S
通讯作者：
Blome S
DOI：
10.3390/v13091731
发表时间：
2021-08-31
期刊：
Viruses
影响因子：
--
作者：
Ceruti A;Kobialka RM;Ssekitoleko J;Okuni JB;Blome S;Abd El Wahed A;Truyen U
通讯作者：
Truyen U
影响因子：
1
作者：
Shimetani, Naoto
通讯作者：
Shimetani, Naoto
影响因子：
4.3
作者：
Diem Hong Tran;Hau Thi Tran;Van Phan Le
通讯作者：
Van Phan Le
影响因子：
13.6
作者：
Bernstein AS;Ando AW;Loch-Temzelides T;Vale MM;Li BV;Li H;Busch J;Chapman CA;Kinnaird M;Nowak K;Castro MC;Zambrana-Torrelio C;Ahumada JA;Xiao L;Roehrdanz P;Kaufman L;Hannah L;Daszak P;Pimm SL;Dobson AP
通讯作者：
Dobson AP