Whole genome phylogenetic analysis in this study resolved a total of five major genotypes among the 22 varicella-zoster virus (VZV) strains or isolates for which complete genomic sequences are available. Consistent with earlier publications we have designated these genotypes European 1 (E1), European 2 (E2), Japanese (J), mosaic 1 (M1), and mosaic 2 (M2). Single nucleotide polymorphism (SNP) analysis performed in a whole-genome alignment revealed that VZV isolates of all five genotypes can be accurately genotyped using SNPs from two amplicons: open reading frame 22 (ORF22) and either ORF21 or ORF50. This modified approach identifies all of the genotypes observed using any of the published genotyping protocols. Of 165 clinical varicella and zoster isolates from Australia and New Zealand typed using this approach, 67 of 127 eastern Australian isolates were E1, 30 were E2, 16 were J, 10 were M1, and 4 were M2; 25 of 38 New Zealand isolates were E1, 8 were E2, and 5 were M1. VZV strain diversity in eastern Australia is thus broader than has been described for any other region, including Europe, Africa, and North America. J strains were far more prevalent than previously observed in countries other than Japan. Two-amplicon typing was in complete accord with genotypes derived using SNP in multiple ORFs (ORFs 1, 21, 22, 38, 50, 54, and 62). Two additional minor genotypes, M3 and M4, could also be resolved using two-amplicon typing.Varicella (chickenpox) results from primary infection with varicella-zoster virus (VZV), which characteristically occurs early in life and usually follows a benign course (1). A lifelong latent infection is established on first exposure and can reactivate, typically after age 50, to cause zoster (shingles). VZV can be transmitted to susceptible persons from either disease condition, although zoster carries a significantly lower risk of transmission. Transmission from zoster can reintroduce strains that were circulating decades earlier and, as such, likely contributes to the genetic stability of VZV. The epidemiology of VZV infection varies geographically. Varicella displays a marked seasonality (peaking in late winter or spring) in temperate climates, and infection is nearly ubiquitous by age 20. For reasons that remain unclear, seasonality does not occur in tropical countries, and a larger proportion of people enter adulthood uninfected by VZV (14, 15). In addition, several studies have demonstrated a distinctive geographic distribution of the major VZV genotypes aligning with cool versus warm global climates (2,18,28). It is unclear whether the strain distribution is actually driven primarily by climate or by other factors, such as immigration patterns.A number of methods have been reported for identifying and genotyping VZV strains (4,18,21). Early VZV typing efforts relied on DNA restriction fragment length polymorphism (RFLP) analysis, an approach that first demonstrated the high degree of sequence conservation among VZV strains. Nevertheless, some intrastrain variation among wild-type i...

Phylogenetic analysis of 19 complete VZV genomic sequences resolves wild-type strains into 5 genotypes (E1, E2, J, M1, and M2). Complete sequences for M3 and M4 strains are unavailable, but targeted analyses of representative strains suggest they are stable, circulating VZV genotypes. Sequence analysis of VZV isolates identified both shared and specific markers for every genotype and validated a unified VZV genotyping strategy. Despite high genotype diversity no evidence for intra-genotypic recombination was observed. Five of seven VZV genotypes were reliably discriminated using only four single nucleotide polymorphisms (SNP) present in ORF22, and the E1 and E2 genotypes were resolved using SNP located in ORF21, ORF22 or ORF50. Sequence analysis of 342 clinical varicella and zoster specimens from 18 European countries identified the following distribution of VZV genotypes: E1, 221 (65%); E2, 87 (25%); M1, 20 (6%); M2, 3 (1%); M4, 11 (3%). No M3 or J strains were observed.

Thirty-one isolates from France and Spain were genotyped using a published method analyzing DNA sequence variation in open reading frame (ORF) 22, together with an evaluation of three well-characterized single nucleotide polymorphisms (SNP) in ORF 38, 54, and 62. Nineteen were allocated to the European (E) genotype, six were mosaic-1 (M1), and two were mosaic-2 (M2). Four strains were assigned to a new genotype, mosaic-4 (M4). All isolates were wild type, with no Oka vaccine-associated markers. No isolates of the mosaic-3 (M3) or Japanese (J) genotype were observed. We also evaluated 13 selected isolates of E, J, M1, and M2 strains (9 of the 31 described above) using an alternative genotyping method based on the assessment of multiple SNP located in ORF 1, 9, 10, 21, 31, 50, 54, 62, and 68. This method assigns wild-type varicella-zoster virus (VZV) strains to seven genotypes: A1, A2, J1, B1, B2, C, and C1. VZV isolates identified as E (ORF22 method) had the genetic signature of genotype C VZV strains, M1 strains were A1, and M2 were A2. No J strains were detected, but parental Oka and vaccine Oka (genotype J) corresponded to genotype J1. M4 isolates (B) share the SNP array observed for M1 and E viruses, and probably represent recombinants between African-Asian (M1) and European (E) viruses. The two genotyping methods, using entirely different genomic targets, produced identical clusters for the strains examined, suggesting robust phylogenetic linkages among VZV strains circulating in Europe.