Diferenciace vakcinačních a divokých kmenů varicella- zoster viru (VZV) pomocí molekulárně biologických metod Diferenciace vakcinačních a divokých kmenů varicella- zoster viru (VZV) pomocí molekulárně biologických metod RNDr. Vanda Boštíková, Ph.D.

Fylogenetická analýza založená na genotypizaci celých sekvencí VZV Loparev et al., J. Virol., Vol. 81, 2007

(rubella)

Loparev et al., J. Virol., Vol. 78, 2004 Globální distribuce VZV - prevalence 70 % a více ve vyznačených oblastech ( E1, E2, M1 – M4, J)

VZV Oka live-attenuated vaccine developed 1974 in Japan by Takahashi and others by extensive passage of a clinical isolate named “Oka” through hu and non-hu cells in vitro A series of clinical trials ( healthy adults and children) demonstrated the safety and efficacy of the vaccine in the USA in 1995: approx. 85% of individuals had complete protection Immunity persists for at least 20 years Healthy People 2020 goals % VZV USA

Varicella in Vaccinated Persons Should we worry about it?

Gomi et al., J. Virol., Vol. 76, 2002

Pst-,Bgl- reported from Australia Preliminary genotyping: The implementation of routine universal varicella vaccination in the US created a need to distinguish wild-type infections from vaccine injuries. Our lab developed LightCycler real-time PCR hybridization probe-based assays to detect VZV(PCR) and genotype VZV strains based on the presence or absence of a BglI, PstI, or MspI restriction sites within open reading frames (ORFs) 54, 38, and 62, respectively.

Pst+ Bgl+ Pst-Bgl+=OKAv Pst+Bgl- Pst+Bgl+ 20% Pst+Bgl+ Pst-Bgl+ Pst+Bgl- 37%

The differences in the melting temperature of the Sma I ORF62 PCR product discriminate Oka vaccine and wt VZV strains.

VZV genotyp ORF 22 : 4 SNPs #37902,38055,38081,38177 European Clade A or D Mosaic Clade C M1 M2 M3 M4 Japanese Clade B + ORF 50/ORF 21 #87841/#3725 Clade A (E1) Genotype like DumasC/T Clade D (E2) T/C

Loparev et al., Virology, 2008

Virology.Virology Jan 20;383(2): Epub 2008 Nov 20. Distribution of varicella-zoster virus (VZV) wild-type genotypes in northern and southern Europe: evidence for high conservation of circulating genotypes. Loparev VNLoparev VN, Rubtcova EN, Bostik V, Tzaneva V, Sauerbrei A, Robo A, Sattler- Dornbacher E, Hanovcova I, Stepanova V, Splino M, Eremin V, Koskiniemi M, Vankova OE, Schmid DS.Rubtcova ENBostik VTzaneva VSauerbrei ARobo ASattler- Dornbacher EHanovcova IStepanova VSplino MEremin VKoskiniemi M Vankova OESchmid DS

Marlene Deleon-Carnes Marlene Deleon-Carnes Antonio Gonzalez Antonio Gonzalez Myron Levin Myron Levin Stephanie Liffick Stephanie Liffick Vladimir Loparev Vladimir Loparev Stanislav Plíšek Stanislav Plíšek Miloslav Salavec Miloslav Salavec Scott D. Schmid Scott D. Schmid