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R esearch
(Published online:
16-07-2015)
10.
Molecular identification and genetic
diversity of open reading frame 7 field isolated porcine
reproductive and respiratory syndrome in North Sumatera,
Indonesia, in the period of 2008-2014 - Faisal Faisal,
Rini Widayanti, Aris Haryanto and Charles Rangga Tabu
Veterinary World, 8(7): 875-880
doi:
10.14202/vetworld.2015.875-880
Faisal
Faisal:
Department of Veterinary Science, Faculty of Veterinary Medicine,
Gadjah Mada University, Yogyakarta, Indonesia; Department of
Molecular Biology, Animal Disease Investigation Centre of Medan,
North Sumatera, Indonesia; faisal.dvm@gmail.com
Rini
Widayanti:
Department of Biochemistry, Faculty of Veterinary Medicine, Gadjah
Mada University, Yogyakarta, Indonesia; riniwida@yahoo.co.uk
Aris
Haryanto:
Department of Biochemistry, Faculty of Veterinary Medicine, Gadjah
Mada University, Yogyakarta, Indonesia; arisharyanto@yahoo.com
Charles
Rangga Tabu: Department of Pathology, Faculty of Veterinary
Medicine, Gadjah Mada University, Yogyakarta, Indonesia;
charles@ugm.ac.id
Received: 11-03-2015, Revised: 13-06-2015, Accepted: 22-06-2015,
Published online: 16-07-2015
Corresponding author:
Faisal Faisal, e-mail: faisal.dvm@gmail.com
Citation:
Faisal F, Widayanti R,
Haryanto A, Tabu CR (2015) Molecular identification and genetic
diversity of open reading frame 7 field isolated porcine
reproductive and respiratory syndrome in North Sumatera,
Indonesia, in the period of 2008-2014, Veterinary World 8(7):
875-880.
Abstract
Aim:
Molecular identification and genetic diversity of open reading
frame 7 (ORF7) of field isolated porcine reproductive and
respiratory syndrome virus (PRRSV) in North Sumatera, Indonesia,
in the period of 2008-2014.
Materials and Methods: A total of 47 PRRSV samples were
collected from the death case of pigs. The samples were collected
from different districts in the period of 2008-2014 from North
Sumatera province. Two pairs of primer were designed to amplify
ORF7 of Type 1 and 2 PRRSV based on the sequence of reference
viruses VR2332 and Lelystad. Viral RNAs were extracted from
samples using PureLink™ micro-to-Midi total RNA purification
system (Invitrogen). To amplify the ORF7 of PRRSV, the synthesis
cDNA and DNA amplification were performed by reverse transcription
polymerase chain reaction (RT-PCR) and nested PCR method. Then the
DNA sequencing of PCR products and phylogenetic analysis were
accomplished by molecular evolutionary genetics analysis version
6.0 software program.
Results: RT-PCR and nested PCR used in this study had
successfully detected of 18 samples positive PRRS virus with the
amplification products at 703bp and 508bp, respectively.
Sequencing of the ORF7 shows that 18 PRRS viruses isolated from
North Sumatera belonged to North American (NA). JXA1 Like and
classic NA type viruses. Several mutations were detected,
particularly in the area of nuclear localization signal (NLS1) and
in NLS2. In the local viruses, which were related closed to JXA1
virus; there are two differences in amino acids in position 12 and
43 of ORF7. Our tested viruses showed that the amino acid
positions 12 and 43 are Asparagine and Arginine, while the
reference virus (VR2332, Lelystad, and JXA1) occupied both by
Lysine. Based on differences in two amino acids at position 12 and
43 showed that viruses from North Sumatera has its own uniqueness
and related closed to highly pathogenic PRRS (HP-PRRS) virus
(JXA1).
Conclusion: The results demonstrated that North Sumatera type
PRRS virus has caused PRRS outbreaks in pig in North Sumatera
between 2008 and 2014. The JAX1 like viruses had unique amino acid
residue in position 12 and 43 of asparagine and lysine, and these
were genetic determinants of North Sumatera viruses compared to
other PRRS viruses.
Keywords: Indonesia, North Sumatera, open
reading frame 7, porcine reproductive and respiratory syndrome
virus.
References
1. Neumann, E.J., Kliebenstein, J.B., Johnson, C.D., Mabry,
J.W., Bush, E.J., Seitzinger, A.H., Gree, A.L. and Zimmerman,
J.J. (2005) Assessment of the economic impact of porcine
reproductive and respiratory syndrome on swine production in
the United States. J. Am. Vet. Med. Assoc., 227: 385-392.
http://dx.doi.org/10.2460/javma.2005.227.385
PMid:16121604 |
|
|
2. Benfield, D.A., Nelson, E., Collins, J.E., Harris, L.,
Goyal, S.M., Robison, D., Christianson, W.T., Morrison, R.B.,
Gorcyca, D. and Chladek, D. (1992) Characterization of swine
infertility and respiratory syndrome (SIRS) virus (isolate
ATCC VR-2332). J. Vet. Diagn. Invest., 4: 127-133.
http://dx.doi.org/10.1177/104063879200400202
PMid:1616976 |
|
|
3. Wensvoort, G., Terpstra, C., Pol, J.M., Ter Laak, E.A.,
Bloemraad, M., de Kluyver, E.P., Kragten, C., van Buiten, L.,
den Besten, A. and Wagenaar, F. (1991) Mystery swine disease
in the Netherlands: The isolation of Lelystad virus. Vet. Q.,
13(3): 121-30.
http://dx.doi.org/10.1080/01652176.1991.9694296
PMid:1835211 |
|
|
4. Tian, K., Yu, X., Zhao, T., Feng, Y., Cao, Z., Wang, C., Hu,
Y., Chen, X., Hu, D., Tian, X., Liu, D., Zhang, S., Deng, X.,
Ding, Y., Yang, L., Zhang, Y., Xiao, H., Qiao, M., Wang, B.,
Hou, L., Wang, X., Yang, X., Kang, L., Sun, M., Jin, P., Wang,
S., Kitamura, Y., Yan, J. and Gao, G.F. (2007) Emergence of
fatal PRRSV variants: Unparalleled outbreaks of atypical PRRS
in Cina and molecular dissection of the unique hallmark. PLoS
One, 2: 526. Available from: http://www.plosone.org.
http://dx.doi.org/10.1371/journal.pone.0000526
PMid:17565379 PMCid:PMC1885284 |
|
|
5. Zhou, L. and Yang, H. (2010), Porcine reproductive and
respiratory syndrome in China. Virus Res., 154: 31-37.
http://dx.doi.org/10.1016/j.virusres.2010.07.016
PMid:20659506 |
|
|
|
6. FAO. (2011) Focus on: Porcine reproductive and respiratory
syndrome (PRRS) virulence jumps and persistent circulation in
Southeast Asia. EMPRES, ???(5): ???. 1-8. |
|
|
|
7. Helen, R., Papadopoulou, C., Drew, T., Gresham, A. and
Sabirovic, M. (2009) Highly pathogenic porcine reproductive
and respiratory syndrome. Int Dis Monit Situat Assess, 23: 10. |
|
|
8. Nilubol, D., Tripipat, T., Hoonsuwan, T. and Kortheerakul,
K. (2012) Porcine reproductive and respiratory syndrome virus,
Thailand, 2010-2011. Emerg. Infect. Dis., 18(12): 2039-2043.
http://dx.doi.org/10.3201/eid1812.111105
PMid:23171619 PMCid:PMC3557868 |
|
|
9. Dokland, T. (2010) The structural biology of PRRSV. Virus
Res., 154: 86-97.
http://dx.doi.org/10.1016/j.virusres.2010.07.029
PMid:20692304 |
|
|
10. Meulenberg, J.J., Petersen-den Besten, A., De Kluyver,
E.P., Moormann, R.J., Schaaper, W.M. and Wensvoort, G. (1995)
Characterization of proteins encoded by ORFs 2 to 7 of
Lelystad virus. Virology, 206: 155-163.
http://dx.doi.org/10.1016/S0042-6822(95)80030-1 |
|
|
11. Yoo, D., Wootton, S.K., Li, G., Song, C. and Rowland, R.R.
(2003) Colocalization and interaction of the porcine
arterivirus nucleocapsid protein with the small nucleolar RNA
associated protein fibrillarin. J. Virol., 77: 12173-12183.
http://dx.doi.org/10.1128/JVI.77.22.12173-12183.2003
PMid:14581554 PMCid:PMC254285 |
|
|
12. Lin, C.N., Lin, W.H., Hung, L.N., Wang, S.Y. and Chiou,
M.T. (2013) Comparison of viremia of Type 2 porcine
reproductive and respiratory syndrome virus in naturally
infected pigs by zip nucleic acid probe-based real-time PCR.
BMC Vet. Res., 9: 181.
http://dx.doi.org/10.1186/1746-6148-9-181
PMid:24028493 PMCid:PMC3847877 |
|
|
13. Wernike, K., Hoffmann, B., Dauber, M., Lange, E.,
Schirrmeier, H. and Beer, M. (2012) Detection and typing of
highly pathogenic porcine reproductive and respiratory
syndrome virus by multiplex real-time RT-PCR. PLoS One, 7(6):
Available from: http://www.plosone.org. Last accessed on
25-01-2015.
http://dx.doi.org/10.1371/journal.pone.0038251 |
|
|
14. Hao, X., Lu, Z., Kuang, W., Sun, P., Fu, Y., Wu, L., Zhao,
Q., Bao, H., Fu, Y., Cao, Y., Li, P., Bai, X., Li, D. and Liu,
Z. (2011) Polymorphic genetic characterization of the ORF7
gene of porcine reproductive and respiratory syndrome virus
(PRRSV) in China. J. Virol., 8: 73.
http://dx.doi.org/10.1186/1743-422X-8-73
PMid:21333014 PMCid:PMC3049123 |
|
|
15. Shi, Y., Hu, Z., Xiong, Z., Zhou, Y., Jin, X., Gu, C., Hu,
X., Cheng, G., Song, N. and Zhang, W. (2013) Analysis of
molecular variation of porcine reproductive and respiratory
syndrome virus in Central China from 2006 to 2012. Arch.
Virol., 158: 717-721.
http://dx.doi.org/10.1007/s00705-012-1542-1
PMid:23160706 |
|
|
16. Guarino, H., Goyal, S.M., Murtaugh, M.P., Morrison, R.B.,
Kapur, V. (1999) Detection of porcine reproductive and
respiratory syndrome virus by reverse transcription–polymerase
chain reaction using different regions of the viral genome. J.
Vet. Diagn. Invest., 11: 27-33.
http://dx.doi.org/10.1177/104063879901100104
PMid:9925208 |
|
|
17. Rowland, R.R. and Yoo, D. (2003) Nucleolar-cytoplasmic
shuttling of PRRSV nucleocapsid protein: A simple case of
molecular mimicry or the complex regulation by nuclear import,
nucleolar localization and nuclear export signal sequences.
Virus Res, 95: 23-33.
http://dx.doi.org/10.1016/S0168-1702(03)00161-8 |
|
|
18. Music, N. and Gagnon, C.A. (2010) The role of porcine
reproductive and respiratory syndrome (PRRS) virus structural
and non-structural proteins in pathogenesis. Anim. Health Res.
Rev., 11(2): 135-163.
http://dx.doi.org/10.1017/S1466252310000034
PMid:20388230 |
|
|
19. Wootton, S.K., Rowland, R.R. and Yoo, D. (2002)
Phosphorylation of the porcine reproductive and respiratory
syndrome virus nucleocapsid protein. J. Virol., 76:
10569-10576.
http://dx.doi.org/10.1128/JVI.76.20.10569-10576.2002
PMid:12239338 PMCid:PMC136587 |
|
|
20. Wootton, S.K. and Yoo, D. (2003) Homo-oligomerization of
the porcine reproductive and respiratory syndrome virus
nucleocapsid protein and the role of disulfide linkages. J.
Virol., 77: 4546-4557.
http://dx.doi.org/10.1128/JVI.77.8.4546-4557.2003
PMid:12663761 PMCid:PMC152152 |
|
|
21. Hu, S.P., Zhang, Z., Liu, Y.G., Tian, Z.J., Wu, D.L., Cai,
X.H. and He, X.J. (2012) Pathogenicity and distribution of
highly pathogenic porcine reproductive and respiratory
syndrome virus in pigs. Transbound Emerg. Dis., 60: 351-359.
http://dx.doi.org/10.1111/j.1865-1682.2012.01354.x
PMid:22762447 |
|
|
22. Metwallly, S., Mohamed, F., Faaberg, K., Burrage, T.,
Prarat, M., Moran, K., Bracht, A., Mayr, G., Berninger, M.,
Koster, L., To, T.L., Nguyen, V.L., Reising, M., Landgraf, J.,
Cox, L., Lubroth, J. and Carrillo, C. (2010) Pathogenicity and
molecular characterization of emerging porcine reproductive
and respiratory syndrome virus in vietnam in 2007. Transbound
Emerg. Dis., 57: 315-329.
http://dx.doi.org/10.1111/j.1865-1682.2010.01152.x
PMid:20629970 |
|
|
23. Rossow, K.D., Collins, J.E., Goyal, S.M., Nelson, E.A.,
Christopher-Hennings, J. and Benfield, D.A. (1995)
Pathogenesis of porcine reproductive and respiratory syndrome
virus infection in gnotobiotic pigs. Vet. Pathol., 32:
361-373.
http://dx.doi.org/10.1177/030098589503200404
PMid:7483210 |
|
|
24. Zhou, Z., Ni, J., Cao, Z., Han, X., Xia, Y., Zi, Z., Ning,
K., Liu, Q., Cai, L., Qiu, P., Deng, X., Hu, D., Zhang, Q.,
Fan, Y., Wu, J., Wang, L., Zhang, M., Yu, X., Zhai, X. and
Tian, K. (2011) The epidemic status and genetic diversity of
14 highly pathogenic porcine reproductive and respiratory
syndrome virus (HP-PRRSV) isolates from China in 2009. Vet.
Microbiol., 150(3-4): 257-269.
http://dx.doi.org/10.1016/j.vetmic.2011.02.013
PMid:21411250 |
|
