[논문]Evaluation of different molecular methods for detection of Senecavirus A and the result of the antigen surveillance in Korea during 2018

Heo, JinHwa; Lee, Min-Jung; Kim, HyunJoo; Lee, SuKyung; Choi, Jida; Kang, Hae-Eun; Nam, Hyang-Mi; Nah, JinJu

doi:10.7853/kjvs.2021.44.1.15

Evaluation of different molecular methods for detection of Senecavirus A and the result of the antigen surveillance in Korea during 2018 원문보기

韓國家畜衛生學會誌 = Korean journal of veterinary service, v.44 no.1, 2021년, pp.15 - 19

Heo, JinHwa (Foreign Animal Disease Division, Animal and Plant Quarantine Agency) , Lee, Min-Jung (Foreign Animal Disease Division, Animal and Plant Quarantine Agency) , Kim, HyunJoo (Foreign Animal Disease Division, Animal and Plant Quarantine Agency) , Lee, SuKyung (Foreign Animal Disease Division, Animal and Plant Quarantine Agency) , Choi, Jida (Foreign Animal Disease Division, Animal and Plant Quarantine Agency) , Kang, Hae-Eun (Foreign Animal Disease Division, Animal and Plant Quarantine Agency) , Nam, Hyang-Mi (Foreign Animal Disease Division, Animal and Plant Quarantine Agency) , Nah, JinJu (Foreign Animal Disease Division, Animal and Plant Quarantine Agency)

Abstract ▼ AI-Helper

Senecavirus A (SVA), previously known as Seneca Valley virus, can cause vesicular disease and neonatal losses in pigs that is clinically indistinguishable from foot-and-mouth disease virus (FMDV). After the first case report in Canada in 2007, it had been restrictively identified in North America including United States. But, since 2015, SVA emerged outside North America in Brazil, and also in several the Asian countries including China, Thailand, and Vietnam. Considering the SVA occurrence in neighboring countries, there has been a high risk that Korea can be introduced at any time. In particular, it is very important in terms of differential diagnosis in the suspected case of vesicular diseases in countries where FMD is occurring. So far, several different molecular detection methods for SVV have been published but not validated as the reference method, yet. In this study, seven different molecular methods for detecting SVA were evaluated. Among them, the method by Flowler et al, (2017) targeted to 3D gene region with the highest sensitivity and no cross reaction with other vesicular disease agents including FMDV, VSV and SVD, was selected and applied further to antigen surveillance of SVA. A total of 245 samples of 157 pigs from 61 farms submitted for animal disease diagnose nationwide during 2018 were tested all negative. In 2018, no sign of SVA occurrence have been confirmed in Korea, but the results of the surveillance for SVA needs to be continued and accumulated at a high risk of SVA in neighboring countries.

주제어

표/그림 (6)

표 Table 1. Conventional RT-PCR (RT-PCR) primers for detecting SVA
표 Table 2. Real-time RT-PCR (rRT-PCR) primers and probes for detecting SVA
그림 Fig. 1. Genome structure of Seneca Valley Virus.
표 Table 3. The results of sensitivity test
표 Table 4. The results of specificity test
표 Table 5. The result of the antigen surveillance result to SVV in Korea during 2018

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제안 방법

Therefore, it is necessary to set-up rapid and accurate diagnostic methods such as PCR for preparing the outbreak of SVA, in advance. In this study, we evaluated seven different molecular methods for detecting SVA and then applied the selected method to antigen surveillance of pig samples in Korea during 2018. As the results of evaluation, the rRT-PCR method by Flowler et al (2017) with the highest sensitivity and no cross reaction with other vesicular disease agents including FMDV was selected and applied to antigen surveillance of SVA in Korea.
For the specificity test, the vesicular disease agents including FMDV, VSV and SVDV were used. Seven different methods of conventional RT-PCR were performed using One-step RT-PCR kit (Qiagen) on Eppendorf Master cycler and Real-time RT-PCR were performed using AgPath-ID one-step RT- PCR kit (Ambion) or Power SYBR Green RNA-to-Ct 1-step kit (Life Tech) on the Bio-Rad CFX96 system, respectively. The information of seven different methods were described in Table 1, 2 in detail.
So far, several different molecular detection methods for SVV have been published but not validated as the reference method, yet. Therefore, in this study, we tried to evaluate several different molecular methods for detecting SVA and then apply the selected method to the antigen surveillance to pig samples in Korea collected during 2018.

대상 데이터

For the sensitivity test, limits of detection (LOD) were estimated 0by 10-fold serial dilution of synthetic RNA to 10 cop- ies/µL, and by 10-fold serial dilution of viral RNA5starting from 3×10 TCID50/mL of SVV001 strain from ATCC PTA-5343, respectively. For the specificity test, the vesicular disease agents including FMDV, VSV and SVDV were used. Seven different methods of conventional RT-PCR were performed using One-step RT-PCR kit (Qiagen) on Eppendorf Master cycler and Real-time RT-PCR were performed using AgPath-ID one-step RT- PCR kit (Ambion) or Power SYBR Green RNA-to-Ct 1-step kit (Life Tech) on the Bio-Rad CFX96 system, respectively.

이론/모형

In this study, we evaluated seven different molecular methods for detecting SVA and then applied the selected method to antigen surveillance of pig samples in Korea during 2018. As the results of evaluation, the rRT-PCR method by Flowler et al (2017) with the highest sensitivity and no cross reaction with other vesicular disease agents including FMDV was selected and applied to antigen surveillance of SVA in Korea. In addition, because the rRT-PCR method by Flower et al (2017) targets conserved 3D re- gion, it has the advantage of being able to more reliably detect SVA belonging to the Picrornaviridae family with high mutation rate.

참고문헌 (17)

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