EuFMD OS20

1. GENETIC CHARACTERIZATION OF SEROTYPE AND GENETIC RELATEDNESS OF
FOOT-AND-MOUTH DISEASE VIRUSES IN SOUTH EAST ASIA
HyunJi Lee1, BooMi La1, Jeong-won Lee1, DongSook Lee1, Jae-Myung Kim1, Nguyen Dang Tho2, Bounlom Douangngeun3,
Seng Bunnary4, Sang-Ho Cha1, Da-Rae Lim*1
1 Animal and Plant Quarantine Agency, Ministry of Agriculture, Food and Rural affairs, Republic of Korea
2 National Center for Veterinary diagnostics, Department of Animal Health, Ministry of Agriculture and Rural Development, Vietnam
3 National Animal Health Laboratory, Department of Livestock and Fisheries, Ministry of Agriculture and Forestry, LAO PDR
4 National Animal Health and Production Research Institute, General Directorate of Animal Health and Production, Kingdom of Cambodia
This research was supported by the Research of Animal and Plant Quarantine Agency, Ministry of
Agriculture, Food and Rural Affairs (MAFRA), Republic of Korea.
References :
VP. Le et al., 2016. BMC Vet Res 12:269
VP. Le et al., 2010. Vet. Microbiol. 45:220-229
RESULTSINTRODUCTION
Figure 2. 95 clinical samples were collected that occurred between 2018 and 2019 from
Vietnam, Laos and Cambodia. Viral RNAs were extracted from the clinical samples and
cells using a Nextracter(Genolution). The 3D region in the virus genome was
confirmed by qRT-PCR to determine whether the collected samples were FMDV
positive. The VP1 regions were amplified using a one-step RT-PCR kit(Qiagen). PCR
products were purified with ExoSAP-IT (USB) and directly sequenced on an ABI 3130
genetic analyzer(Applied Biosystems) using a Big Dye Terminator Kit v3.1(Applied
Biosystems). VP1 genome sequences were alinged by BioEdit and analyzed
Phylogenetic tree estimated using the Maximum-Likelihood method on the General
Time Reversible model in MEGA-X version 10.0.1.
Foot-and-mouth disease (FMD) is the most important disease of livestock, causing huge
economic impact on worldwide. FMD virus is a single strand of positive-sense RNA and
over 8,000 bases in length. The causative agent, FMD virus (FMDV), is divided into seven
Serotype categories: A, O, C, Asia1, SAT1, SAT2 and SAT3 based on VP1 sequence.
Each serotype is divided into several topotypes and lineages by genetic
characterization. FMDV serotype O and A are the most prevalent in Southeast Asia
containing Vietnam, Laos, Cambodia, and Myanmar. Two different lineages of serotype O
have been circulating in Southeast Asia during 2018-2019; O/ME-SA/PanAsia and
O/SEA/Mya-98. And A/ASIA/SEA97 of serotype A also have been circulating in Southeast
Asia. In Korea, FMDV serotype O/ME-SA/PanAsia was reported in 2000 and 2002.
O/SEA/Mya-98 was reported in 2010, 2014 and 2016, and A/ASIA/SEA97 was reported in
2010 and 2018. With the recent increase in physical and human exchanges with
Southeast Asia, the possibility of introducing the FMDV serotype O/ME-SA/PanAsia into
South Korea is increasing. In this study, we collected 95 clinical FMDV samples between
2018 and 2019 in Vietnam, Laos and Cambodia, analyzed genetic relatedness.
Figure 1. A) Genome structure of Foot-and-mouth disease virus (FMDV) FMDV is a
non-enveloped, single-stranded positive-sense RNA virus from the genus
Aphthovirus within the family Picornaviridae which rapidly spreads among cloven-
footed animals. B) Seven types of FMDVs; O, A, C, Asia1, and Southern African
Territories(SAT) 1 to 3. C) The spreading of FMDV from Southeast Asia into South
Korea As exchanges between South Korea and Southeast Asia have increased, the
likelihood that viruses from Southeast Asia will spread to South Korea will increase.
D) In South Korea, 11 foot-and-mouth disease cases have occurred since 2000, and
only serotype O and A have occurred.
A
B
C
MATERIALS AND METHODS
ACKNOWLEDGEMENT & REFERENCES
A B
Figure 3. A) It was possible to determine whether FMD was positive or negative through
3D qRT-PCR. As a result of analyzing 95 FMDV samples, 40 were found to be FMD
positive. B) Major capsid protein, VP1 were amplified using RT-PCR. Amplified PCR
products were separated by gel electrophoresis, stained with ethidium bromide.
FMDV from Vietnam, Laos, Cambodia
(3D qRT-PCR)
CONCLUSION
D
FMDVs in Vietnam and Laos belonged to Serotype O and classified into O/Me-
SA/Panasia and O/SEA/Mya-98 genotypes. FMDVs in Cambodia belonged to Serotype O
and A, being classified into O/ME-SA/Panasia and A/ASIA/SEA-97 genotype. The further
study may require investigation of antigenic relatedness of the field FMDVs to strains of
commercial FMD vaccines. Also, Securing FMDV in a Pool 1 region and accumulating
the genetic information of the virus can provide valuable information for further research
on the genetic diversity and antigenic relationship of FMDV to prevent and control foot-
and-mouth disease in South Korea in the future.
Phylogenetic Tree
Serotyping A
A/AISA/SEA-97
CAMBODIA
O/ME-SA/Panasia
VIETNAM, LAOS
CAMBODIA
O/SEA/Mya98
VIETNAM
Phylogenetic Tree
Serotyping O
Figure 4. Phylogenetic analysis of FMD positive samples from Vietnam, LAO, and
Cambodia. As a result of genome analysis based on the VP1 gene, Cambodia's
A/ASIA/SEA-97 and Vietnam's O/SEA/Mya98 each belong to one genetic cluster. Some
FMDVs in Cambodia and Vietnam and FMDV in Laos (n=1) belong to three different
genetic clusters in O/ME-SA/Panasia.