Enzymatic Digestion Pattern of Varicella Zoster Virus ORF38 and ORF54 in Chickenpox Patients Using RFLP Technique

Document Type: Original Research

Authors

1 Dept of Virology, Iran University of Medical Sciences, Tehran, Iran.

2 Gastrointestinal amp; Liver Disease Research Center (GILDRC), Firuzgar Hospital, Tehran, Iran

Abstract

Background: Varicella zoster virus (VZV) causes chickenpox in children and zoster (zona) in the elderly. Using RFLP-PCR method for detection of VZV specific SNPs ORF38, 54 and 62 could  distinguish the profile of VZV isolates. The aim of this study was to investigate enzymatic digestion pattern of VZV ORF38 and ORF54 in chickenpox patients using RFLP technique.
Methods:
Thirty-eight chickenpox patients, who referred to the hospitals of Iran University of Medical Sciences in Tehran from May 2010 to June 2015 were enrolled in this cross sectional study. After the DNA extraction, PCR amplification of 38 VZV isolates performed by specific primers of ORFs 38 and 54, then RFLP assay and digestion carried out by PstI (for ORF38) and BglI (for ORF54) restriction enzymes.
Results:
Of 38 positive VZV DNA, the mean age (yr)±SD was 34.4±23.3 (range: 7-89). 22 (57.9%) were female and 16 (42.1%) were male. The predominant VZV profile of BglI+PstI+ were 89.5% (34/38) followed by 10.5% (4/38) PstI+BglI‾. Statistical analysis showed that there was no significant relationship between genotype, age, sex, and year of infection variables (P value> 0.05). The common VZV genotype among Iranian patients with chickenpox and zona infection is genotype BglI+PstI+ followed by PstI+BglI‾.
Conclusion:
There are different VZV circulating genotypes that call for for more research on this field by widely population and other methods such as nucleotide sequencing to justify the accurate VZV genotype prevalence in Iran.

Keywords


  1. Breuer J, Grose C, Norberg P, Tipples G, Schmid DS. A proposal for a common nomenclature for viral clades that form the species varicella-zoster virus: summary of VZV Nomenclature Meeting 2008, Barts and the London School of Medicine and Dentistry, 24-25 July 2008. J Gen Virol 2010;91(Pt 4):821-8.
  2. Depledge DP, Gray ER, Kundu S, Cooray S, Poulsen A, Aaby P, et al. Evolution of cocirculating varicella-zoster virus genotypes during a chickenpox outbreak in Guinea-Bissau. J Virol 2014;88(24):13936-46.
  3. Brisson M, Edmunds WJ, Law B, Gay NJ, Walld R, Brownell M, et al. Epidemiology of varicella zoster virus infection in Canada and the United Kingdom - CORRIGENDUM. Epidemiol Infect 2014;1:232-40.
  4. Kawai K, Gebremeskel BG, Acosta CJ. Systematic review of incidence and complications of herpes zoster: towards a global perspective. BMJ Open 2014;4(6):e004833.
  5. Mahshid M, Baharak A, Iraj S, Sina K, Javad K, Mehdi B. Seroprevalence of canine visceral leishmaniasis in southeast of Iran. J Parasit Dis 2014;38(2):218-22.
  6. Loparev VN, McCaustland K, Holloway BP, Krause PR, Takayama M, Schmid DS. Rapid genotyping of varicella-zoster virus vaccine and wild-type strains with fluorophore-labeled hybridization probes. J Clin Microbiol 2000;38(12):4315-9.
  7. Salleras L, Dominguez A, Vidal J, Plans P, Salleras M, Taberner JL. Seroepidemiology of varicella-zoster virus infection in Catalonia (Spain). Rationale for universal vaccination programmes. Vaccine 2000;19(2-3):183-8.
  8. Chow VT, Tipples GA, Grose C. Bioinformatics of varicella-zoster virus: single nucleotide polymorphisms define clades and attenuated vaccine genotypes. Infect Genet Evol. 2013;18:351-6.
  9. Weinert LA, Depledge DP, Kundu S, Gershon AA, Nichols RA, Balloux F, et al. Rates of vaccine evolution show strong effects of latency: implications for Varicella Zoster virus epidemiology. Mol Biol Evol 2015;8:133-9.
  10. Sauerbrei A, Wutzler P. Different genotype pattern of varicella-zoster virus obtained from patients with varicella and zoster in Germany. J Med Virol 2007;79(7):1025-31.
  11. Norberg P, Tyler S, Severini A, Whitley R, Liljeqvist JA, Bergstrom T. A genome-wide comparative evolutionary analysis of herpes simplex virus type 1 and varicella zoster virus. PLoS One 2011;6(7):e22527.
  12. Campsall PA, Au NH, Prendiville JS, Speert DP, Tan R, Thomas EE. Detection and genotyping of varicella-zoster virus by TaqMan allelic discrimination real-time PCR. J Clin Microbiol 2004;42(4):1409-13.
  13. Roycroft E, Rose L, Scallan MF, Crowley B. Molecular characterization of varicella-zoster virus clinical isolates from 2006 to 2008 in a tertiary care hospital, Dublin, Ireland, using different genotyping methods. J Med Virol 2012;84(10):1672-9.
  14. Faga B, Maury W, Bruckner DA, Grose C. Identification and mapping of single nucleotide polymorphisms in the varicella-zoster virus genome. Virology 2001;280(1):1-6.
  15. Muir WB, Nichols R, Breuer J. Phylogenetic analysis of varicella-zoster virus: evidence of intercontinental spread of genotypes and recombination. J Virol 2002;76(4):1971-9.
  16. Loparev VN, Rubtcova EN, Bostik V, Govil D, Birch CJ, Druce JD, et al. Identification of five major and two minor genotypes of varicella-zoster virus strains: a practical two-amplicon approach used to genotype clinical isolates in Australia and New Zealand. J Virol 2007;81(23):12758-65.
  17. Sauerbrei A, Zell R, Philipps A, Wutzler P. Genotypes of varicella-zoster virus wild-type strains in Germany. J Med Virol 2008;80(6):1123-30.
  18. Hashemilar M, Ghabili K, Shoja MM, Savadi-Oskouei D, Keyvani H. Varicella-zoster virus reactivation from multiple ganglia: a case report. J Med Case Rep 2009;3:9134-8.
  19. Pourahmad M, Davami MH, Jahromi AR. Evaluation of anti-varicella antibody in young women before their marriage: A sero-epidemiologic study in Iran. J Clin Virol 2010;48(4):260-3.
  20. Schmidt-Chanasit J, Sauerbrei A. Evolution and world-wide distribution of varicella-zoster virus clades. Infect Genet Evol 2011;11(1):1-10.
  21. Loparev VN, Gonzalez A, Deleon-Carnes M, Tipples G, Fickenscher H, Torfason EG, et al. Global identification of three major genotypes of varicella-zoster virus: longitudinal clustering and strategies for genotyping. J Virol 2004;78(15):8349-58.
  22. Kim KH, Choi YJ, Song KH, Park WB, Jeon JH, Park SW, et al. Genotype of varicella-zoster virus isolates in South Korea. J Clin Microbiol 2011;49(5):1913-6.
  23. Lohitharajah J, Malavige GN, Wijewickrama A, Ambagawita A, Seneviratne AL, Ogg G. Molecular characterisation of varicella zoster virus genotypes in Sri Lanka. Ceylon Med J 2013;58(4):153-6.
  24. Quinlivan M, Hawrami K, Barrett-Muir W, Aaby P, Arvin A, Chow VT, et al. The molecular epidemiology of varicella-zoster virus: evidence for geographic segregation. J Infect Dis 2002;186(7):888-94.
  25. Loparev VN, Rubtcova EN, Bostik V, Tzaneva V, Sauerbrei A, Robo A, et al. Distribution of varicella-zoster virus (VZV) wild-type genotypes in northern and southern Europe: evidence for high conservation of circulating genotypes. Virology 2009;383(2):216-25.
  26. Jiang L, Gan L, Chen J, Wang M. Genetic analysis of clinical VZV isolates collected in China reveals a more homologous profile. Biomed Res Int 2013;2013:681234.