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009.4 Estimating hsv-2 superinfection using a novel custom genotyping platform
  1. C Johnston1,2,
  2. A Magaret2,3,4,8,
  3. C Sather4,
  4. K Diem2,
  5. M Huang2,
  6. S Selke2,
  7. JR Lingappa1,5,6,
  8. C Celum1,5,7,
  9. DM Koelle1,2,4,5,
  10. A Wald1,2,4,7
  1. 1Department of Medicine, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
  2. 2Laboratory Medicine, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
  3. 3Biostatistics, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
  4. 4University of Washington, Vaccine and Infectious Disease Division, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
  5. 5Global Health, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
  6. 6Pediatrics, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
  7. 7Epidemiology, Fred Hutchinson Cancer Research Center, Seattle, WA, USA
  8. 8Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

Abstract

Introduction Quantitative estimation of the protective effect of HSV-2 infection against reinfection with other HSV-2 strains is an important parameter for HSV-2 vaccine development. We determined the prevalence of and risk factors for HSV-2 superinfection using a novel genotyping tool.

Methods We first identified 96 high quality HSV SNPs that could determine whether HSV-2 strains were matched with >90% probability via next generation sequencing of 39 genital HSV-2 lesion swabs. These SNPs were then used to create a customised high throughput genotyping assay (GoldenGate, Illumina®). Two genital specimens collected from the same participant, each containing ≥5 log10 copies HSV DNA/ml, were genotyped. HIV-infected and HIV-uninfected persons participating in studies in the USA, Africa, and Peru were included. Sample pairs were excluded if <90% SNP calls were valid. Participants were considered to be infected with more than one strain of HSV-2 if their samples differed by ≥3 SNPs between the paired samples.

Results Paired genital swab specimens from 123 persons were analysed; 113 (92%) had the same strain detected at the two time points; 93 (76%) had identical SNP patterns, 18 (15%) had disagreements at one SNP, and 2 (2%) had disagreements at 2 SNPs. Ten persons (8%) were infected with more than one strain, with paired samples disagreeing at a median of 23 SNPs (range 5–33), for a minimum estimated superinfection prevalence of 8%. Of the 10 persons with HSV-2 superinfection, 7 (70%) were women and 7 (70%) were HIV infected; 6 were from Africa, one was from the USA, and 3 were from Peru.

Conclusion We developed a custom genotyping assay that provides a high throughput method for genotyping HSV-2. HSV-2 superinfection was detected in 8% of paired samples, suggesting that naturally occurring immunity to HSV-2 may not be highly efficient to protect against reinfection, especially among HIV-infected persons.

Disclosure of interest statement This study was funded by the US National Institutes of Health. No pharmaceutical grants were received for the conduct of this study.

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