期刊论文详细信息
Virology Journal
Molecular and phylogenetic analysis of the porcine kobuvirus VP1 region using infected pigs from Sichuan Province, China
Wen-yu Yang1  Wan-zhu Guo1  Zhi-wen Xu1  Yuan-cheng Zhou1  Ling Zhu1  Lei Chen1 
[1] Animal Biotechnology Center, College of Veterinary Medicine of Sichuan Agricultural University, 46# Xinkang Road, Yucheng District, Ya’an 625014Sichuan province, P.R. China
关键词: Recombination;    Co-infection;    Phylogenetic analysis;    Diarrhea;    VP1 protein;    Porcine kobuvirus;   
Others  :  821627
DOI  :  10.1186/1743-422X-10-281
 received in 2013-07-09, accepted in 2013-09-10,  发布年份 2013
PDF
【 摘 要 】

Background

Porcine kobuvirus (PKoV) is a member of the Kobuvirus genus within the Picornaviridae family. PKoV is distributed worldwide with high prevalence in clinically healthy pigs and those with diarrhea.

Methods

Fecal and intestinal samples (n = 163) from pig farms in Sichuan Province, China were obtained to determine the presence of PKoV using reverse transcription polymerase chain reaction assays. Specific primers were used for the amplification of the gene encoding the PKoV VP1 protein sequence. Sequence and phylogenetic analyses were conducted to clarify evolutionary relationships with other PKoV strains.

Results

Approximately 53% (87/163) of pigs tested positive for PKoV. PKoV was widespread in asymptomatic pigs and those with diarrhea. A high prevalence of PKoV was observed in pigs younger than 4 weeks and in pigs with diarrhea. Phylogenetic analysis of 36 PKoV VP1 protein sequences showed that Sichuan PKoV strains formed four distinct clusters. Two pigs with diarrhea were found to be co-infected with multiple PKoV strains. Sequence and phylogenetic analyses revealed diversity within the same host and between different hosts. Significant recombination breakpoints were observed between the CHN/SC/31-A1 and CHN/SC/31-A3 strains in the VP1 region, which were isolated from the same sample.

Conclusion

PKoV was endemic in Sichuan Province regardless of whether pigs were healthy or suffering from diarrhea. Based on our statistical analyses, we suggest that PKoV was the likely causative agent of high-mortality diarrhea in China from 2010. For the first time, we provide evidence for the co-existence of multiple PKoV strains in one pig, and possible recombination events in the VP1 region. Our findings provide further insights into the molecular properties of PKoV, along with its epidemiology.

【 授权许可】

   
2013 Chen et al.; licensee BioMed Central Ltd.

【 预 览 】
附件列表
Files Size Format View
20140712081831741.pdf 707KB PDF download
Figure 3. 85KB Image download
Figure 2. 82KB Image download
Figure 1. 111KB Image download
【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】
  • [1]Reuter G, Boldizsár Á, Pankovics P: Complete nucleotide and amino acid sequences and genetic organization of porcine kobuvirus, a member of a new species in the genus Kobuvirus, family Picornaviridae . Arch Virol 2009, 154:101-108.
  • [2]Yamashita T, Ito M, Kabashima Y, Tsuzuki H, Fujiura A, Sakae K: Isolation and characterization of a new species of kobuvirus associated with cattle . J Gen Virol 2003, 84:3069-3077.
  • [3]Yamashita T, Kobayashi S, Sakac K, Nakata S, Chiba S, Ishihara Y, Isomura S: Isolation of cytopathic small round viruses with BS-Cl cells from patients with gastroenteritis . J Infect Dis 1991, 164:954-957.
  • [4]Reuter G, Boldizsár Á, Kiss I, Pankovics P: Candidate new species of Kobuvirus in porcine hosts . Emerg Infect Dis 1968, 2008:14.
  • [5]Reuter G, Boros Á, Pankovics P, Egyed L: Kobuvirus in domestic sheep . Hungary. Emerg Infect Dis 2010, 16:869.
  • [6]Reuter G, Nemes C, Boros Á, Kapusinszky B, Delwart E, Pankovics P: Porcine kobuvirus in wild boars (Sus scrofa) . Arch Virol 2013, 158:281-282.
  • [7]Kapoor A, Simmonds P, Dubovi EJ, Qaisar N, Henriquez JA, Medina J, Shields S, Lipkin WI: Characterization of a canine homolog of human Aichivirus . J Virol 2011, 85:11520-11525.
  • [8]Lee M, Jeoung H, Lim J, Song J, Song D, An D: Kobuvirus in South Korean black goats . Virus genes 2012, 45:186-189.
  • [9]Carmona-Vicente N, Buesa J, Brown PA, Merga JY, Darby AC, Stavisky J, Sadler L, Gaskell RM, Dawson S, Radford AD: Phylogeny and prevalence of kobuviruses in dogs and cats in the UK . Vet Microbiol 2013, 164:246-252.
  • [10]Li L, Victoria JG, Wang C, Jones M, Fellers GM, Kunz TH, Delwart E: Bat guano virome: predominance of dietary viruses from insects and plants plus novel mammalian viruses . J Virol 2010, 84:6955-6965.
  • [11]Phan TG, Kapusinszky B, Wang C, Rose RK, Lipton HL, Delwart EL: The fecal viral flora of wild rodents . PLoS pathogens 2011, 7:e1002218.
  • [12]Yamashita T, Sakae K, Tsuzuki H, Suzuki Y, Ishikawa N, Takeda N, Miyamura T, Yamazaki S: Complete Nucleotide sequence and genetic organization of Aichi virus, a distinct member of the Picornaviridae associated with acute gastroenteritis in humans . J Virol 1998, 72:8408-8412.
  • [13]Pham NTK, Khamrin P, Nguyen TA, Kanti DS, Phan TG, Okitsu S, Ushijima H: Isolation and molecular characterization of Aichi viruses from fecal specimens collected in Japan, Bangladesh, Thailand, and Vietnam . J Clin Microbiol 2007, 45:2287-2288.
  • [14]Oh D, Silva PA, Hauroeder B, Diedrich S, Cardoso D, Schreier E: Molecular characterization of the first Aichi viruses isolated in Europe and in South America . Arch Virol 2006, 151:1199-1206.
  • [15]Ambert-Balay K, Lorrot M, Bon F, Giraudon H, Kaplon J, Wolfer M, Lebon P, Gendrel D, Pothier P: Prevalence and genetic diversity of Aichi virus strains in stool samples from community and hospitalized patients . J Clin Microbiol 2008, 46:1252-1258.
  • [16]Sdiri-Loulizi K, Gharbi-Khelifi H, De Rougemont A, Chouchane S, Sakly N, Ambert-Balay K, Hassine M, Guédiche MN, Aouni M, Pothier P: Acute infantile gastroenteritis associated with human enteric viruses in Tunisia . J Clin Microbiol 2008, 46:1349-1355.
  • [17]Khamrin P, Maneekarn N, Peerakome S, Okitsu S, Mizuguchi M, Ushijima H: Bovine kobuviruses from cattle with diarrhea . Emerg Infect Dis 2008, 14:985.
  • [18]Reuter G, Egyed L: Bovine kobuvirus in Europe . Emerg Infect Dis 2009, 15:822.
  • [19]Barry AF, Ribeiro J, Alfieri AF, van der Poel WH, Alfieri AA: First detection of kobuvirus in farm animals in Brazil and the Netherlands . Infect Genet Evol 2011, 11:1811-1814.
  • [20]Yu J, Jin M, Zhang Q, Li H, Li D, Xu Z, Li J, Cui S, Yang S, Liu N: Candidate porcine kobuvirus . China. Emerg Infect Dis 2009, 15:823.
  • [21]Khamrin P, Maneekarn N, Kongkaew A, Kongkaew S, Okitsu S, Ushijima H: Porcine kobuvirus in piglets, Thailand . Emerg Infect Dis 2009, 15:2075-2076.
  • [22]Halaihel N, Fuertes H, de Blas I, Gironés O: Kobuvirus and hepatitis-E virus infections in piglets: epidemiology and possible zoonosis . Virology 2011, 3:134.
  • [23]Khamrin P, Maneekarn N, Hidaka S, Kishikawa S, Ushijima K, Okitsu S, Ushijima H: Molecular detection of kobuvirus sequences in stool samples collected from healthy pigs in Japan . Infect Genet Evol 2010, 10:950-954.
  • [24]An D, Jeoung H, Jeong W, Lee HS, Park J, Kim B: Porcine kobuvirus from pig stool in Korea . Virus genes 2011, 42:208-211.
  • [25]Verma H, Mor SK, Abdel-Glil MY, Goyal SM: Identification and molecular characterization of porcine kobuvirus in US swine . Virus genes 2013, 46:551-553.
  • [26]Dufkova L, Scigalkova I, Moutelikova R, Malenovska H, Prodelalova J: Genetic diversity of porcine sapoviruses, kobuviruses, and astroviruses in asymptomatic pigs: an emerging new sapovirus GIII genotype . Arch Virol 2012, 158:549-558.
  • [27]Reuter G, Kecskeméti S, Pankovics P: Evolution of porcine kobuvirus infection . Hungary. Emerg Infect Dis 2010, 16:696.
  • [28]Okitsu S, Khamrin P, Thongprachum A, Hidaka S, Kongkaew S, Kongkaew A, Maneekarn N, Mizuguchi M, Hayakawa S, Ushijima H: Sequence analysis of porcine kobuvirus VP1 region detected in pigs in Japan and Thailand . Virus genes 2012, 44:253-257.
  • [29]Park S, Kim H, Moon H, Song D, Rho S, Han J, Nguyen V, Park B: Molecular detection of porcine kobuviruses in pigs in Korea and their association with diarrhea . Arch Virol 2010, 155:1803-1811.
  • [30]Yu J, Xu Z, Li B, Zhang Q, Cui S, Jin M, Duan Z: Analysis and characterization of the complete genome of a member of a new species of kobuvirus associated with swine . Arch Virol 2011, 156:747-751.
  • [31]Wang C, Lan D, Hua X: Porcine kobuvirus from pig stool specimens in Shanghai, China . Virus genes 2011, 43:350-352.
  • [32]Reuter G, Boros Á, Pankovics P: Kobuviruses–a comprehensive review . Rev Med Virol 2011, 21:32-41.
  • [33]Shi D, Zhang S, Chen J, Shi H, Zhang X, Feng L: Molecular characterization of a porcine kobuvirus variant strain in China . Arch Virol 2013.
  • [34]Padidam M, Sawyer S, Fauquet CM: Possible emergence of new geminiviruses by frequent recombination . Virology 1999, 265:218-225.
  • [35]Martin DP, Posada D, Crandall KA, Williamson C: A modified bootscan algorithm for automated identification of recombinant sequences and recombination breakpoints . AIDS Res Hum Retrov 2005, 21:98-102.
  • [36]Smith JM: Analyzing the mosaic structure of genes . J Mol Evol 1992, 34:126-129.
  • [37]Posada D, Crandall KA: Evaluation of methods for detecting recombination from DNA sequences: computer simulations . P Natl Acad Sci 2001, 98:13757-13762.
  • [38]Gibbs MJ, Armstrong JS, Gibbs AJ: Sister-scanning: a Monte Carlo procedure for assessing signals in recombinant sequences . Bioinformatics 2000, 16:573-582.
  • [39]Lole KS, Bollinger RC, Paranjape RS, Gadkari D, Kulkarni SS, Novak NG, Ingersoll R, Sheppard HW, Ray SC: Full-length human immunodeficiency virus type 1 genomes from subtype C-infected seroconverters in India, with evidence of intersubtype recombination . J Virol 1999, 73:152-160.
  • [40]Pham NTK, Trinh QD, Khamrin P, Nguyen TA, Dey SK, Phan TG, Hoang LP, Maneekarn N, Okitsu S, Mizuguchi M: Sequence analysis of the capsid gene of Aichi viruses detected from Japan, Bangladesh, Thailand, and Vietnam . J Med Virol 2008, 80:1222-1227.
  • [41]Lau SK, Woo PC, Yip CC, Li KS, Fu CT, Huang Y, Chan K, Yuen K: Co-existence of multiple strains of two novel porcine bocaviruses in the same pig, a previously undescribed phenomenon in members of the family Parvoviridae, and evidence for inter-and intra-host genetic diversity and recombination . J Gen Virol 2011, 92:2047-2059.
  • [42]Han X, Zhang W, Xue Y, Shao S: Sequence analysis reveals mosaic genome of Aichi virus . Virol J 2011, 8:390. BioMed Central Full Text
  • [43]Lukashev AN: Recombination among picornaviruses . Rev Med Virol 2010, 20:327-337.
  • [44]Park S, Kim H, Song D, Moon H, Park B: Molecular detection and genetic characterization of kobuviruses in fecal samples collected from diarrheic cattle in Korea . Infect Genet Evol 2011, 11:1178-1182.
  • [45]Yamashita T, Sugiyama M, Tsuzuki H, Sakae K, Suzuki Y, Miyazaki Y: Application of a reverse transcription-PCR for identification and differentiation of Aichi virus, a new member of the Picornavirus family associated with gastroenteritis in humans . J Clin Microbiol 2000, 38:2955-2961.
  • [46]Le Guyader FS, Le Saux J, Ambert-Balay K, Krol J, Serais O, Parnaudeau S, Giraudon H, Delmas G, Pommepuy M, Pothier P: Aichi virus, norovirus, astrovirus, enterovirus, and rotavirus involved in clinical cases from a French oyster-related gastroenteritis outbreak . J Clin Microbiol 2008, 46:4011-4017.
  • [47]Cao W, Zheng H, Zhang K, Jin Y, Lv L, Yang F, Liu X: Complete genome sequence of the porcine kobuvirus variant CH/HNXX-4/2012 . J Virol 2012, 86:11947.
  • [48]Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S: MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods . Mol Biol Evol 2011, 28:2731-2739.
  • [49]Martin D, Rybicki E: RDP: detection of recombination amongst aligned sequences . Bioinformatics 2000, 16:562-563.
  文献评价指标  
  下载次数:13次 浏览次数:9次