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Volume 12, Number 10—October 2006
Dispatch

Fourth Human Parechovirus Serotype

Kimberley S.M. Benschop*Comments to Author , Janke Schinkel*, Manon E. Luken†, Peter J.M. van den Broek†, Matthias F.C. Beersma‡, Negassi Menelik§, Hetty W.M. van Eijk*, Hans L. Zaaijer*, Christina M.J.E. VandenBroucke-Grauls*, Marcel G.H.M. Beld*, and Katja C. Wolthers*
Author affiliations: *Academic Medical Center, Amsterdam, the Netherlands; †Primagen, Amsterdam, the Netherlands; ‡Leiden University Medical Center, Leiden, the Netherlands; §BovenIJ Ziekenhuis, Amsterdam, the Netherlands

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Table 2

Neutralization assay with LLcMk2 cells*

Virus Antiserum Viral controls
a-HPeV1 (Harris) a-HPeV2 (Williamson) a-HPeV3 (A308-99)
HPeV1 Harris – ++++ ++++ ++++
HPeV2 Williamson ++++ – ++++ ++++
K251181-02 (HPeV3) ++++ ++++ – ++++
K251176-02 ++++ ++++ ++++ ++++

*Culture isolates of K251176-02, human parechovirus 1 (HPeV1, echovirus 22) and HPeV2 (echovirus 23) from a reference panel (National Institute for Public Health and the Environment, Bilthoven, the Netherlands) and K251181-02 that was previously genotyped as HPeV3 (12) were incubated with antisera (20 U/mL in Eagle minimal essential medium) directed against HPeV1 Harris, HPeV2 Williamson, and HPeV3 A308-99. The antisera to HPeV1 and HPeV2 were raised in horses. The antiserum to HPeV3 was raised in guinea pigs. Neutralization is done on a 96-microtiter plate containing a monolayer of LLcMk2 cells that have been incubated for 3 days. The assay was determined after viral controls (no antisera used) of the 4 culture isolates showed cytopathic effects >50% (++++).

*Culture isolates of K251176-02, human parechovirus 1 (HPeV1, echovirus 22) and HPeV2 (echovirus 23) from a reference panel (National Institute for Public Health and the Environment, Bilthoven, the Netherlands) and K251181-02 that was previously genotyped as HPeV3 (12) were incubated with antisera (20 U/mL in Eagle minimal essential medium) directed against HPeV1 Harris, HPeV2 Williamson, and HPeV3 A308-99. The antisera to HPeV1 and HPeV2 were raised in horses. The antiserum to HPeV3 was raised in guinea pigs. Neutralization is done on a 96-microtiter plate containing a monolayer of LLcMk2 cells that have been incubated for 3 days. The assay was determined after viral controls (no antisera used) of the 4 culture isolates showed cytopathic effects >50% (++++).

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