Complete genome sequencing and analysis of six enterovirus 71 strains with different clinical phenotypes

Virology Journal

Zhi-yu Wang³  Cheng-xi Sun³  Fu-lu Chu³  Feng Gao¹  Shu-bin Hao²  Xiao-jing Yuan³  Lu-ying Si³  Hong-ling Wen³ 

[1] Linyi People's Hospital, Linyi, Shandong, 276000, People’s Republic of China;Shandong Medical Equipment Quality Supervision And Inspection Center, Jinan, Shandong, 250012, People’s Republic of China;The Key Laboratory of experimental teratology of Ministry of Education, Department of Virology, School of Public Health, Shandong University, Jinan, Shandong, 250012, People’s Republic of China

关键词: Hand, foot and mouth disease;    Virulent determinant;    Enterovirus 71;   

Others  :  1150923
DOI  :  10.1186/1743-422X-10-115

 received in 2012-06-14, accepted in 2013-04-02,  发布年份 2013

【 摘 要 】

Background

Hand, foot and mouth diseases (HFMD) caused by enterovirus 71(EV71) presents a broad spectrum of clinical manifestations ranging from mild febrile disease to fatal neurolocal disease. However, the mechanism of virulence is unknown.

Methods

We isolated 6 strains of EV71 from HFMD patients with or without neurological symptoms, and sequenced the whole genomes of the viruses to reveal the virulence factors of EV71.

Results

Phylogenetic tree based on VP1 region showed that all six strains clustered into C4a of C4 sub-genotype. In the complete polypeptide, 298 positions were found to be variable in all strains, and three of these positions (Val^P814/Ile^P814 in VP1, Val^P1148/Ile^P1148 in 3A and Ala ^P1728/Cys ^P1728/Val ^P1728 in 3C) were conserved among the strains with neurovirulence, but variable in strains without neurovirulence. In the 5^′-UTR region, it showed that the first 10 nucleotides were mostly conserved, however from the 11th nucleotide, nucleotide insertions and deletions were quite common. The secondary structure prediction of 5^′-UTR sequences showed that two of three strains without neurovirulence (SDLY11 and SDLY48) were almost the same, and all strains with neurovirulence (SDLY96, SDLY107 and SDLY153) were different from each other. SDLY107 (a fatal strain) was found different from other strains on four positions (C^P241/T^P241, A^P571/T^P571, C^P579/T^P579 in 5^′-UTR and T^P7335/C^P7335 in 3^′-UTR).

Conclusions

The three positions (Val^P814/Ile^P814 in VP1, Val^P1148/Ile^P1148 in 3A and Ala ^P1728/Cys ^P1728/Val ^P1728 in 3C), were different between two phenotypes. These suggested that the three positions might be potential virulent positions. And the three varied positions were also found to be conserved in strains with neurovirulence, and variable in strains without neurovirulence. These might reveal that the conservation of two of the three positions or the three together were specific for the strains with neurovirulence. Varation of secondary structure of 5^′-UTR, might be correlated to the changes of viral virulence. SDLY107 (a fatal strain) was found different from other strains on four positions, these positions might be related with death.

【 授权许可】

   
2013 Wen et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Hagiwara A, Yoneyama T, Takami S, Hashimoto I: Genetic and phenotypic characteristics of enterovirus 71 isolates from patients with encephalitis and with hand, foot and mouth disease. Arch Virol 1984, 79(3–4):273-283.
• [2]Blomberg J, Lycke E, Ahlfors K, Johnsson T, Wolontis S, von Zeipel G: New enterovirus type associated with epidemic of aseptic meningitis and or hand, foot, and mouth disease. Lancet 1974, 2(7872):112.
• [3]Chumakov M, Voroshilova M, Shindarov L, Shindarov L, Lavrova I: Enterovirus 71 isolated from cases of epidemic poliomyelitis-like disease in Bulgaria. Arch Virol 1979, 60(3–4):329-340.
• [4]Ishimaru Y, Nakano S, Yamaoka K, Takami S: Outbreaks of hand, foot, and mouth disease by enterovirus 71. High incidence of complication disorders of central nervous system. Arch Dis Child 1980, 55(8):583-588.
• [5]Lan YC, Lin TH, Tsai JD, Yang YC, Peng CT: Molecular epidemiology of the 2005 enterovirus 71 outbreak in central Taiwan. Scand J Infect Dis 2011, 43(5):345-349.
• [6]Kim KH: Enterovirus 71 infection: An experience in Korea 2009. Korean J Pediatr 2010, 53(5):616-622.
• [7]Li Wei A, Benjamin KW K, Kwai Peng C, Chua LT, James L, Goh KT: Epidemiology and Control of Hand,Foot and Mouth Disease in Singapore, 2001–2007. Ann Acad Med Singapore 2009, 38(2):106-112.
• [8]Schuffenecker I, Mirand A, Antona D, Henquell C, Chomel JJ: Epidemiology of human enterovirus 71 infections in France, 2000–2009. J Clin Virol 2011, 50(1):50-56.
• [9]McMinn P, Stratov I, Nagarajan L, Davis S: Neurological manifestations of enterovirus 71 infection in children during an outbreak of hand, foot, and mouth disease in Western Australia. Clin Infect Dis 2001, 32(2):236-242.
• [10]Chan LG, Parashar UD, Lye MS, Ong FG, Zaki SR: Deaths of children during an outbreak of hand, foot, and mouth disease in sarawak, malaysia: clinical and pathological characteristics of the disease. For the Outbreak Study Group. Clin Infect Dis 2000, 31(3):678-683.
• [11]Komatsu H, Shimizu Y, Takeuchi Y, Ishiko H, Takada H: Outbreak of severe neurologic involvement associated with enterovirus 71 infection. Pediatr Neuro 1999, 20(1):17-23.
• [12]Zheng ZM, He PJ, Caueffield D, Neumann M, Specter S: Enterovirus 71 isolated from China is serologically similar to the prototype E71 BrCr strain but differs in the 5′-noncoding region. J Med Virol 1995, 47(2):161-167.
• [13]Chang SC, Li WC, Chen GW, Tsao KC, Huang CG: Genetic characterization of enterovirus 71 isolated from patients with severe disease by comparative analysis of complete genomes. J Med Virol 2012, 84(6):931-939.
• [14]Melchers WJ, Hoenderop JG, Bruins Slot HJ, Pleij CW, Pilipenko EV: Kissing of the two predominant hairpin loops in the coxsackie B virus 3′untranslated region is the essential structural feature of the origin of replication required for negative-strand RNA synthesis. J Virol 1997, 71(1):686-696.
• [15]Wimmer E, Hellen CU, Cao X: Genetics of poliovirus. Annu Rev Genet 1993, 27:353-436.
• [16]Bedard KM, Semler BL: Regulation of picornavirus gene expression. Microbes Infect 2004, 6(7):702-713.
• [17]Pelletier J, Kaplan G, Racaniello VR, Sonenberg N: Cap-independent translation of poliovirus mRNA is conferred by sequence elements within the 5′noncoding region. Mol Cell Biol 1988, 8(3):1103-1112.
• [18]Liu XL, Wang ZG, Yang TT, Yi T: Molecular epidemiology of human enterovirus 71 strains in Qingdao region, Shandong province, 2007–2009. Chin J Epidemiol 2011, 32(4):382-384.
• [19]Evans DM, Dunn G, Minor PD, Schild GC, Cann AJ: Increased neurovirulence associated with a single nucleotide change in a noncoding region of the Sabin type 3 poliovaccine genome[J]. Nature 1985, 314(6011):548-550.
• [20]Guillot S, Otelea D, Delpeyroux F, Crainic R: Point mutations involved in the attenuation/neurovirulence alternation in type 1 and 2 oral polio vaccine strains detected by site-specific polymerase chain reaction. Vaccine 1994, 12(6):503-507.
• [21]Rezapkin GV, Fan L, Asher DM, Fibi MR, Dragunsky EM: Mutations in Sabin 2 strain of poliovirus and stability of attenuation phenotype. Virology 1999, 258(1):152-160.
• [22]Arita M, Shimizu H, Nagata N, Ami Y, Suzaki Y: Temperature-sensitive mutants of enterovirus 71 show attenuation in cynomolgus monkeys. J Gen Virol 2005, 86(5):1391-1401.
• [23]Li R, Zou Q, Chen L, Zhang H, Wang Y: Molecular Analysis of Virulent Determinants of Enterovirus 71. PLoS One 2011, 6(10):e26237.
• [24]Lin JY, Chen TC, Weng KF, Chang SC, Chen LL: Viral and host proteins involved in picornavirus life cycle. J Biomed Sci 2009, 16(1):103. BioMed Central Full Text
• [25]Foo DG, Alonso S, Phoon MC, Ramachandran NP, Chow VT: Identification of neutralizing linear epitopes from the VP1 capsid protein of Enterovirus 71 using synthetic peptides. Virus Res 2007, 125(9):61-68.
• [26]Guidelines for the prevention and treatment of HFMD. http://www.moh.gov.cn/publicfiles/business/cmsresources/wsb/cmsrsdocument/doc4510.doc webcite