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J Gen Virol 84 (2003), 827-836; DOI 10.1099/vir.0.18647-0

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© 2003 Society for General Microbiology

Sequencing of ‘untypable’ enteroviruses reveals two new types, EV-77 and EV-78, within human enterovirus type B and substitutions in the BC loop of the VP1 protein for known types

Helene Norder1, Lotte Bjerregaard1, Lars Magnius1, Bruno Lina2, Michèle Aymard2 and Jean-Jacques Chomel2

1 Swedish Institute for Infectious Disease Control, S-171 82 Solna, Sweden
2 Centre National de Reference des Enterovirus, Lyon, France

Correspondence
Lars Magnius
lars.magnius{at}smi.ki.se

The N-terminal part of VP1 was sequenced for 43 enterovirus isolates that could not initially be neutralized with LBM pools or in-house antisera. Most isolates were found to belong to human enterovirus type A (HEV-A) and HEV-B (18 isolates of each). All HEV-A isolates could be typed by sequencing, with CV (coxsackievirus)-A16 and EV (enterovirus)-71 being dominant (nine and seven isolates, respectively). These types thus seem to have diverged more from their prototypes than the other types. Among the HEV-B isolates, E-18 dominated with five isolates that became typable after filtration. The virus type obtained by molecular typing was verified for 28 of the other patient isolates by neutralization using high-titre monovalent antisera or LBM pools. Twenty-two of the other 30 ‘untypable’ isolates had substitutions in the VP1 protein within or close to the BC loop. Two closely related HEV-B isolates diverged by 19·4 % from E-15, the most similar prototype. Two non-neutralizable HEV-C isolates split off from the CV-A13/CV-A18 branch, from which they diverged by 15·7–18·2 %. Three of the six non-neutralizable isolates, W553-130/99, W543-122/99 and W137-126/99, diverged by >24·2 % from the most similar prototype in the compared region. The complete VP1 was therefore sequenced and found to diverge by >29 % from all prototypes and by >28 % from each other. Strains similar to W553-130/99 that have been identified in the USA are tentatively designated EV-74. The two other isolates fulfil the molecular criterion for being new types. Since strains designated EV-75 and EV-76 have been identified in the USA, we have proposed the tentative designations EV-77 and EV-78 for these two new members of HEV-B.

The complete VP1 sequences of EV-74, EV-77 and EV-78 and the partial VP1 sequences of the other isolates are deposited in GenBank with accession nos AY208081AY208120.




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