RNDr. Vanda Bo štíková , Ph.D .

transcript

Diferenciace Diferenciace vakcinačních a vakcinačních a

divokých kmenů divokých kmenů varicella-zoster viru varicella-zoster viru

(VZV) pomocí (VZV) pomocí molekulárně molekulárně

biologických metodbiologických metod

RNDr. RNDr. Vanda BoVanda Boštíková, Ph.D.štíková, 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

(2011 - 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 (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 yearsHealthy People 2020 goals 2010 90.51% VZV USA

Varicella in Vaccinated Varicella in Vaccinated PersonsPersons

Should we worry about it?Should we worry about it?

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

Genotyping

VaccineWildtype

Japanese

Wildtype

USWildtype

VZVGenotyping

Pst +/-

OKAvWildtypeJapan

Wildtype20% USAWAustraliaAfrica,Asia

WildtypeEurope,EAustralia,80% US

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-

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

VZVgenotyp ORF 22 :

4 SNPs #37902,38055,38081,38177

European Clade A or D

Mosaic Clade CM1M2M3M4

JapaneseClade 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. 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-CarnesMarlene Deleon-Carnes Antonio GonzalezAntonio Gonzalez Myron LevinMyron Levin Stephanie LiffickStephanie Liffick Vladimir LoparevVladimir Loparev Stanislav PlíšekStanislav Plíšek Miloslav SalavecMiloslav Salavec Scott D. Schmid Scott D. Schmid

RNDr. Vanda Bo štíková , Ph.D .

Documents