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Diferenciace vakcinačních a divokých kmenů varicella - zoster viru (VZV) pomocí molekulárně biologických metod. RNDr. Vanda Bo štíková , Ph.D. Fylogenetick á analýza založená na genotypizaci celých sekvencí VZV. Loparev et al., J. Virol., Vol. 81, 2007. (rubella).
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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
Globální distribuce VZV - prevalence 70 % a více ve vyznačených oblastech (2011 - E1, E2, M1 – M4, J) Loparev et al., J. Virol., Vol. 78, 2004
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 (11 000 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 HealthyPeople 2020 goals 2010 90.51% VZV USA
Preliminary genotyping: The implementation of routine universal varicella vaccination in the US created a need to distinguish wild-type infections from vaccine injuries. VZV VZV Genotyping Genotyping Pst Pst - - Pst Pst + + Pst Pst + + Pst Pst +/- - Bgl Bgl + + Bgl Bgl + - Bgl Bgl - + Bgl Bgl + + Msp Msp + + Msp Msp - - Msp Msp - - Msp Msp - - Sma Sma + + Sma Sma - - Sma Sma - - Sma Sma - - Wildtype Japan Wildtype 20% USA WAustralia Africa,Asia Wildtype Europe,EAustralia, 80% US Wildtype Wildtype OKAv Vaccine Wildtype Japanese US Pst-,Bgl- reported from Australia 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- Pst+Bgl- Pst+Bgl- 20% Pst+Bgl+ Pst+ Bgl+ Pst-Bgl+=OKAv Pst+Bgl+ Pst-Bgl+ Pst+Bgl+ 37% Pst+Bgl+ Pst+Bgl+ Pst+Bgl-
The differences in the melting temperature of the Sma I ORF62 PCR product discriminate Oka vaccine and wt VZV strains.
ORF 22 : 4 SNPs #37902,38055,38081,38177 VZV genotyp Mosaic Clade C M1 M2 M3 M4 European Clade A or D Japanese Clade B + Clade A (E1) Genotype like DumasC/T ORF 50/ORF 21 #87841/#3725 Clade D (E2) T/C
Virology. 2009 Jan 20;383(2):216-25. 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 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.
Marlene Deleon-Carnes • Antonio Gonzalez • Myron Levin • Stephanie Liffick • Vladimir Loparev • Stanislav Plíšek • Miloslav Salavec • Scott D. Schmid