Amalia Magaret

University of Washington Seattle, Seattle, Washington, United States

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Publications (78)540.27 Total impact

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    ABSTRACT: To evaluate the utility of quantitative herpes simplex virus (HSV) polymerase chain reaction (PCR) levels for prognosis and management of neonatal HSV disease. Clinical and virologic data were abstracted by medical record review from neonatal HSV cases treated at Seattle Children's Hospital between 1993 and 2012. HSV PCR results from plasma (n = 47), cerebrospinal fluid (n = 56), or both (n = 40) at the time of diagnosis were available from 63 infants; 26 with skin-eye-mouth (SEM), 18 with central nervous system (CNS), and 19 with disseminated (DIS) disease. Plasma HSV PCR was positive in 78% of the infants with SEM, 64% with CNS and 100% with DIS disease. Mean plasma viral level was 2.8 log10 copies/mL in SEM, 2.2 log10 copies/mL in CNS, and 7.2 log10 copies/mL in DIS infants. The HSV levels were higher among infants who died compared with surviving infants, 8.1 log10 copies/mL (range 7.7-8.6) vs 3.8 log10 copies/mL (range 0.0-8.6), P = .001, however, level of HSV DNA in the cerebrospinal fluid or in plasma did not correlate with neurologic outcome. Dynamics of HSV clearance from plasma during high-dose acyclovir treatment showed single-phase exponential decay with a median viral half-life of 1.26 days (range: 0.8-1.51). Plasma HSV levels correlate with clinical presentation of neonatal HSV disease and mortality, but not neurologic outcome. Copyright © 2015 Elsevier Inc. All rights reserved.
    The Journal of pediatrics. 12/2014;
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    ABSTRACT: Background: Genital HSV-2 infection causes recurrent genital lesions and frequent viral shedding; current therapies do not completely control viral replication and require daily antiviral therapy. The therapeutic vaccine GEN003 contains 2 HSV-2 proteins, gD and ICP4, and Matrix M2 adjuvant (MM) to promote both B and T cell immune responses. Methods: We conducted a randomized, placebo controlled clinical trial in which 143 participants with genital HSV-2 infection were randomized to receive 3 vaccinations with 10, 30 or 100 µg of GEN003 with or without MM, or placebo. Participants collected twice daily swabs of genital secretions to assess genital shedding, and recorded genital lesions for 28 days prior to, immediately after; and 6 and 12 months after completion of vaccinations. Safety, immunogenicity and efficacy were monitored. Results: Immediately after vaccination, viral shedding in the 30 µg and 100 µg with MM groups decreased from baseline (rate ratios [RR] 0.47 and 0.68, respectively; p<0.001 for both). Shedding was unchanged from baseline for the placebo (RR= 1.01), unadjuvanted GEN003 (RR= 1.40), and 10 µg with MM groups (RR=1.01). Shedding remained suppressed for the 30 µg with MM dose group at 6 months (RR=0.57; p<0.001). Lesion rates were reduced after vaccination for the 30 and 100 µg with MM dose groups (RR 0.42 and 0.35, respectively; p<0.001 for both) and at 6 months for the 30 µg with MM dose (RR=0.26; p<0.001). In a preliminary data review, at 12 months, lesion rates for the 30 µg with MM dose remained reduced (9.6% [n=29] at baseline versus 2.4% [n=17] at 12 months). Three participants in the 10 µg with MM group discontinued dosing due to adverse events. Injection reactions were previously reported (Wald, ICAAC 2013). Other than reactogenicity, the number (%) of participants who reported 1 or more other Grade 3 or 4 AEs considered related to vaccination by treatment group were: 10 µg with MM, 3 (10%); 30 µg with MM, 2 (7%); 100 µg with MM, 1 (4%); GEN003 without adjuvant 1 (4%); Placebo, 0 (0%). Conclusion: The antiviral effect of GEN003 persisted for at least 6 months with an effect on genital lesions up to 12 months. Immunotherapy with GEN003 may be an effective strategy, with or without traditional antiviral agents, to treat chronic genital HSV-2 infections.
    IDWeek 2014 Meeting of the Infectious Diseases Society of America; 10/2014
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    ABSTRACT: Background: HSV-1 infection has a wide severity spectrum in the immunocompetent host, from asymptomatic seropositivity to frequent orolabial lesions. To gain insight into virus and host genotype contributions to disease phenotype, we evaluated HSV-1 genotypes and immunity in mono- (MZ) and dizygotic (DZ) twins. Methods: HSV-1 seropositive twin pairs collected daily oral swabs for quantitative HSV-1 PCR and kept symptom diaries for 60 days. Associations of shedding rates were assessed by estimating correlations. We categorized the viral strain as identical or different in each pair with DNA available for genotyping from both. The identity and breadth of HSV-1 antigens recognized by circulating CD4 T-cells were determined using a complete HSV-1 ORF (open reading frame) set. CD4 T-cell responses were scored as present or absent to each ORF. We used a bootstrap method to estimate the distribution of agreements in ORF responses between individuals. Results: We enrolled 29 MZ and 22 DZ twin pairs. The overall shedding rate was 10.3% of days (median 9.3%; range 0-47%). There was a positive correlation between shedding rates within twin pairs (r=0.33, p=0.015) but not among unrelated individuals (r=-0.086; p=0.5). Genotyping showed that 15/14 twin pairs had the same/different HSV-1 strain, respectively. The correlation for shedding rates in all twin pairs was higher in those with the same virus (r=0.55, p=0.033) versus different (r=-0.169, p=0.56). 8 MZ pairs were analyzed for CD4 T-cell responses. The median number of ORFs recognized per person was 19 (range 6-35). The bootstrapped mean percent agreement in ORF response between unrelated pairs was 71% (5th/95th percentile, 67%/75% respectively). The percent agreement between the original 8 pairs of MZ twins was 77% (p=0.003 for difference from bootstrapped dataset). Conclusion: A relationship between HSV-1 shedding and host genotype is supported by our observation of a higher correlation in HSV-1 shedding between twin pairs than between unrelated individuals and similar CD4 T-cell responses between MZ twins. These data and the higher correlation in shedding rates among twins with the same versus different virus suggests that both viral and host genetics contribute to HSV-1 severity.
    IDWeek 2014 Meeting of the Infectious Diseases Society of America; 10/2014
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    ABSTRACT: Background: Vaccines are most successful for pathogens in which wild-type infection leads to protection against homologous re-infection. For HSV-2, we have little information concerning natural protection from re-infection or the variability between strains that could modulate protection from challenge. Understanding differences in HSV-2 strains worldwide, and the potential for reinfection, is essential in development of an HSV-2 vaccine. Methods: DNA from genital HSV-2 lesions containing ≥7 log10 copies HSV DNA/swab were sequenced using the Illumina next generation sequencing (NGS) platform. Human and highly repetitive sequences were subtracted, and remaining reads were aligned to the reference HSV-2 genome. Prevalent SNPs were defined as present in >10% and <90% of the sequenced strains. Informative SNPs were identified using FastTagger. Bayesian computations were used to estimate probabilities of identifying super-infection. Results: Fifty-six samples from genital herpes lesions were sequenced. Of 49 sequences from 39 persons with adequate coverage, the HSV genome had a median of 73.9 fold coverage (range 9.8-771 fold). Sixteen persons (41%) were from Africa, 12 (31%) from Peru, and 11 (28%) from the US; one-third were HIV infected. Of 2854 loci with heterogeneity, prevalent SNPs were found at 456 (16%) loci. Using genotyping at the 6 most informative SNPs to distinguish strains, we estimate a 90% probability of correctly identifying super-infection or lack thereof. Of 10 persons with paired specimens collected a median of 2.1 years apart, 2 (20%) had super-infection; both were HIV-infected. In each case, the strains differed at 186 loci. Six of 10 remaining pairs differed at 0 loci, and 2 differed at only 1 locus, indicating minimal within host evolution and low sequencing error. Conclusion: High quality HSV-2 sequence data can be obtained directly from genital swabs, removing the need for culture which may introduce mutations. The initial phase of our study has identified super-infection in 20% of persons investigated. Further research will determine whether host immune status, gender, and sexual exposure history are correlates of super-infection.
    IDWeek 2014 Meeting of the Infectious Diseases Society of America; 10/2014
  • JAMA The Journal of the American Medical Association 08/2014; 312(7):746-8. · 29.98 Impact Factor
  • The Journal of infectious diseases. 06/2014;
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    ABSTRACT: Human Cytomegalovirus (CMV) has historically been the major infectious cause of morbidity and mortality among patients receiving hematopoietic cell or organ transplant. Standard care in a transplant setting involves frequent monitoring of CMV viral load over weeks to months to determine when anti-viral treatment may be required. Quantitative PCR (qPCR) is the standard molecular diagnostic method for monitoring. Recently, digital PCR (dPCR) has shown promise in viral diagnostics, although current dPCR systems have lower throughput than qPCR systems. Here we compare qPCR and droplet digital PCR (ddPCR) for CMV detection in patient plasma samples. Droplet digital PCR exhibits increased precision over qPCR at viral loads ≥ 4 log 10 with equivalent sensitivity. However, retrospective analysis of longitudinal samples from transplant patients with CMV viral loads near therapeutic thresholds did not provide evidence that the improved precision of ddPCR would be of clinical benefit. Given the throughput advantages of current qPCR systems, a widespread switch to dPCR for CMV monitoring would appear premature.
    Journal of clinical microbiology. 05/2014;
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    ABSTRACT: Resiquimod, a toll-like receptor 7 and 8 agonist, stimulates production of cytokines that promote an antigen-specific T helper type 1 acquired immune response. Animal and phase II human trials showed post-treatment efficacy in reducing recurrent herpes lesion days and/or time to first recurrence. Three phase III randomized, double-blind, vehicle-controlled trials of topical resiquimod to reduce anogenital herpes recurrences were conducted in healthy adults with ≥4 recurrences within the prior year. Participants applied resiquimod 0.01% or vehicle gel 2 times per week for 3 weeks to each recurrence for 12 months. Trials 1 and 2 had 2:1 resiquimod:vehicle randomization. Trial 3 had 1:1:1 randomization for resiquimod plus valacyclovir 500 mg orally twice daily for 5 days (RESI/VAL), resiquimod plus oral placebo (RESI/PLA), and vehicle plus oral placebo (VEH/PLA). Median time to first recurrence was similar for resiquimod and vehicle [Trial 1: 60 and 56 days, p=0.7; Trial 2: 54 and 48 days, p=0.47; Trial 3: 51 (RESI/VAL), 55 (RESI/PLA), and 44 (VEH/PLA) days, p=NS]. Median time to healing of initial treated recurrence was longer for resiquimod [Trial 1: 18 versus 10 days, p<0.001; Trial 2: 19 versus 13 days, p=0.16; Trial 3: 14 (RESI/VAL), 16 (RESI/PLA), and 8 (VEH/PLA) days, p<0.001]. In Trials 1 and 2, moderate to severe erythema and erosion/ulceration at the application site were more common in resiquimod recipients. In conclusion, no post-treatment efficacy of resiquimod 0.01% gel was observed. Increased application site reactions and initial recurrence healing time are consistent with resiquimod-induced cytokine effects.
    Antimicrobial Agents and Chemotherapy 04/2014; · 4.57 Impact Factor
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    ABSTRACT: Background. Depot medroxyprogesterone acetate (DMPA) has been linked to HIV-1 acquisition.Methods. Vaginal microbiota of women using DMPA for up to two years were cultured; mucosal immune cell populations measured by immunohistological staining.Results. Over 12 months, the proportion with H2O2-positive lactobacilli decreased (n=32; 53 vs. 27%; p=0.03). Median vaginal CD3(+) cells also decreased (n=15; 355 vs. 237cells/mm(2); p=0.03), as did CD3(+)CCR5(+)(195 vs. 128cells/mm(2); p=0.04), HLA-DR(+) (130 vs. 96cells/mm(2); p=0.27), and HLA-DR(+)CCR5(+) cells (18 vs. 10cells/mm(2); p=0.33).Conclusion. DMPA contraception does not increase vaginal mucosal CCR5(+) HIV-1 target cells, but does decrease CD3(+) T lymphocytes and vaginal H2O2-producing lactobacilli.
    The Journal of Infectious Diseases 03/2014; · 5.85 Impact Factor
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    ABSTRACT: Human herpesvirus 6 (HHV-6) latently infects a majority of adults. In about 1% of the population HHV-6 exists in a chromosomally integrated form (ciHHV-6) that resides in every somatic and germ cell and can be transmitted through the germ line. Patients with ciHHV-6 have been misdiagnosed and unnecessarily treated for active HHV-6 infection, sometimes with important side effects, based on results from quantitative molecular HHV-6 tests.METHODS: A droplet digital PCR (ddPCR) assay was developed to identify ciHHV-6 in cellular patient samples by precisely determining the ratio of HHV-6 to cellular DNA. We validated the assay on confirmed ciHHV-6 patient samples and a cell line derived from a ciHHV-6 patient, and we analyzed hematopoietic stem cell transplant patients suspected of having ciHHV-6. We additionally evaluated whether the assay could be applied to stored plasma samples from a study of clinical correlates of HHV-6.RESULTS: The ddPCR assay accurately identified ciHHV-6 in cellular samples (buffy coat, peripheral blood mononuclear cells), giving a ratio very close to 1 HHV-6/cell [mean (SD), 1.02 (0.03)] in fluorescence in situ hybridization-confirmed samples). In stored plasma samples, the assay performance was set by design to have 100% sensitivity, which resulted in 82% specificity for ciHHV-6.CONCLUSIONS: The possibility of ciHHV-6 is often overlooked in patients with detectable HHV-6 viral loads by quantitative PCR. Our ddPCR test provides rapid and accurate laboratory identification of ciHHV-6 from easily obtained cellular samples. In addition, the assay provides excellent sensitivity and specificity using stored plasma samples, facilitating retrospective analysis of the clinical significance of ciHHV-6.
    Clinical Chemistry 03/2014; · 7.15 Impact Factor
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    ABSTRACT: Genital HSV reactivation is thought to be anatomically and temporally localized, coincident with limited ganglionic infection. Short, subclinical shedding episodes are the most common form of HSV-2 reactivation, with host clearance mechanisms leading to rapid containment. The anatomic distribution of shedding episodes has not been characterized.To precisely define patterns of anatomic reactivation, we divided the genital tract into a 22-region grid and obtained daily swabs for 20 days from each region in 28 immunocompetent, HSV-2 seropositive persons. HSV was detected via PCR and sites of asymptomatic HSV shedding were biopsied within 24 hours. CD4+ and CD8+ T cells were quantified by immunofluorescence, and HSV specific CD4+ T cells were identified by intracellular cytokine cytometry.HSV was detected in 868 (7%) of 11,603 genital swabs at a median of 12 sites per person (range 0-22). Bilateral HSV detection occurred on 83 (67%) days with shedding, and the median quantity of virus detected/day was associated with the number of sites positive (p<0.001). In biopsies of asymptomatic shedding sites, we found increased numbers of CD8+ T cells compared to control tissue (27 vs. 13 cells/mm(2), p=0.03) and identified HSV specific CD4+ T cells.HSV reactivations emanate from widely separated anatomic regions of the genital tract and are associated with a localized cellular infiltrate that was demonstrated to be HSV-specific in 3 cases. These data provide evidence that asymptomatic HSV-2 shedding contributes to chronic inflammation throughout the genital tract. This detailed study of the anatomic patterns of genital HSV-2 shedding demonstrates that HSV-2 reactivation can be detected at multiple, bilateral sites in the genital tract, suggesting that HSV establishes latency throughout the sacral ganglia. In addition, genital biopsies from sites of asymptomatic HSV shedding have increased numbers of CD8+ T cells compared to control tissue, and HSV-specific CD4+ T cells are found at sites of asymptomatic shedding. These findings suggest that that widespread asymptomatic genital HSV-2 shedding is associated with a targeted host immune response and contributes to chronic inflammation throughout the genital tract.
    Journal of Virology 02/2014; · 5.08 Impact Factor
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    ABSTRACT: Pritelivir, an inhibitor of the viral helicase-primase complex, exhibits antiviral activity in vitro and in animal models of herpes simplex virus (HSV) infection. We tested the efficacy and safety of pritelivir in otherwise healthy persons with genital HSV-2 infection. We randomly assigned 156 HSV-2-positive persons with a history of genital herpes to receive one of four doses of oral pritelivir (5, 25, or 75 mg daily, or 400 mg weekly) or placebo for 28 days. Participants obtained daily swabs from the genital area for HSV-2 testing, which was performed with a polymerase-chain-reaction assay. Participants also maintained a diary of genital signs and symptoms. The primary end point was the rate of genital HSV shedding. HSV shedding among placebo recipients was detected on 16.6% of days; shedding among pritelivir recipients was detected on 18.2% of days among those receiving 5 mg daily, 9.3% of days among those receiving 25 mg daily, 2.1% of days among those receiving 75 mg daily, and 5.3% of days among those receiving 400 mg weekly. The relative risk of viral shedding with pritelivir, as compared with placebo, was 1.11 (95% confidence interval [CI], 0.65 to 1.87) with the 5-mg daily dose, 0.57 (95% CI, 0.31 to 1.03) with the 25-mg daily dose, 0.13 (95% CI, 0.04 to 0.38) with the 75-mg daily dose, and 0.32 (95% CI, 0.17 to 0.59) with the 400-mg weekly dose. The percentage of days with genital lesions was also significantly reduced, from 9.0% in the placebo group to 1.2% in both the group receiving 75 mg of pritelivir daily (relative risk, 0.13; 95% CI, 0.02 to 0.70) and the group receiving 400 mg weekly (relative risk, 0.13; 95% CI, 0.03 to 0.52). The rate of adverse events was similar in all groups. Pritelivir reduced the rates of genital HSV shedding and days with lesions in a dose-dependent manner in otherwise healthy men and women with genital herpes. (Funded by AiCuris; ClinicalTrials.gov number, NCT01047540.).
    New England Journal of Medicine 01/2014; 370(3):201-10. · 54.42 Impact Factor
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    ABSTRACT: Following acute infection, herpes simplex virus (HSV) establishes latency in sensory neurons, from which it can reactivate and cause recurrent disease. Available antiviral therapies do not affect latent viral genomes; therefore, they do not prevent reactivation following therapy cessation. One possible curative approach involves the introduction of DNA double strand breaks in latent HSV genomes by rare-cutting endonucleases, leading to mutagenesis of essential viral genes. We tested this approach in an in vitro HSV latency model using the engineered homing endonuclease (HE) HSV1m5, which recognizes a sequence in the HSV-1 gene UL19, encoding the virion protein VP5. Coexpression of the 3'-exonuclease Trex2 with HEs increased HE-mediated mutagenesis frequencies up to sixfold. Following HSV1m5/Trex2 delivery with adeno-associated viral (AAV) vectors, the target site was mutated in latent HSV genomes with no detectable cell toxicity. Importantly, HSV production by latently infected cells after reactivation was decreased after HSV1m5/Trex2 exposure. Exposure to histone deacetylase inhibitors prior to HSV1m5/Trex2 treatment increased mutagenesis frequencies of latent HSV genomes another two- to fivefold, suggesting that chromatin modification may be a useful adjunct to gene-targeting approaches. These results support the continuing development of HEs and other nucleases (ZFNs, TALENs, CRISPRs) for cure of chronic viral infections.Molecular Therapy-Nucleic Acids (2014) 3, e1; doi:10.1038/mtna.2013.75; published online 4 February 2014.
    Molecular therapy. Nucleic acids. 01/2014; 3:e146.
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    ABSTRACT: Background Human herpesvirus 8 (HHV-8) replication increases the risk of Kaposi sarcoma (KS). Highly-active antiretroviral therapy (HAART) reduces the incidence of KS, and regimens that contain protease inhibitors (PIs) may be particularly effective. Objective To determine whether PI-based HAART regimens may more effectively inhibit HHV-8 shedding compared to regimens without PIs. Study design: Prospective, observational study of 142 HIV-1 and HHV-8 co-infected men conducted in Seattle, Washington. Quantitative HHV-8 PCR testing was performed on daily swabs of the oropharynx, the primary site of HHV-8 replication. Associations between antiretroviral regimen and detection of HHV-8 DNA in swabs were evaluated using generalized estimating equations. Results HHV-8 DNA was detected in 3,016 (26%) of 11,608 specimens collected. PI-based HAART was associated with a statistically significantly lower frequency of detection (RR 0.2; 95% CI 0.1 to 0.5) compared to ART-naïve persons, whereas HAART without a PI was not (RR 0.7; 95% CI 0.4 to 1.3). Compared to ART-naïve persons, there was also a trend toward lower quantities of HHV-8 detected during treatment with HAART regimens that contained a PI. These associations between PIs and measures of HHV-8 shedding could not be attributed to use of nelfinavir, which inhibits HHV-8 replication in vitro, and were independent of CD4 count and HIV plasma viral load (VL). Conclusions HAART regimens that contain PIs appear to decrease HHV-8 shedding compared to NNRTIs. Further study of PI-based HAART is warranted to determine the optimal regimens for prevention and treatment of KS.
    Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 01/2014; · 3.12 Impact Factor
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    ABSTRACT: Background. Despite high herpes simplex virus type 2 (HSV-2) incidence and prevalence among women in Africa, we are unaware of published neonatal herpes reports. To assess neonatal HSV transmission potential in South Africa, we investigated the frequency of the strongest risk factors: HSV acquisition in late pregnancy and HSV shedding during labor. Methods. Women admitted in early labor to a hospital in Soweto underwent HSV serologic testing and genital swab collection for HSV PCR. HSV-2 seronegative women were assessed for seroconversion 4-6 weeks after delivery. Results. Of 390 women enrolled, 229 (58.7%) were HSV-2 seropositive. Genital HSV-2 was detected in 17.2% of HSV-2 seropositive women, including 26 of 115 HIV-positive and 13 of 110 HIV-negative women (22.6% versus 11.8%; RR, 1.91; 95% CI, 1.04-3.53; P = 0.038), but in none of 161 HSV-2 seronegative women. Among the 91 HSV-2 seronegative women followed after delivery, none seroconverted. Conclusions. HSV-2 reactivation is common among South African women during labor, especially those with HIV coinfection. To determine the epidemiology of neonatal herpes in South Africa and to investigate whether the lack of reported cases is due to alterations in immune control or HSV-2 virulence, studies evaluating acutely ill neonates for HSV and studies of maternal HSV-2 shedding patterns are needed.
    Infectious Diseases in Obstetrics and Gynecology 01/2014; 2014:258291.
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    ABSTRACT: Human cytomegalovirus (CMV) infection has been implicated in immunosenescence. To examine the influence of CMV on ability of healthy adults to respond to a novel influenza antigen, the rate of seroconversion and the magnitude of titers to pandemic 2009 H1N1 vaccine was assessed. The clinical trial was stratified by age; 52 persons aged 18-64 and 55 aged 65 and older were enrolled. Among the younger group, 33% had CMV antibody compared with 62% among the older group. No differences by CMV seropositivity in the proportion of participants achieving a seroprotective titer 21 days following the second immunization were noted. However, the geometric mean titer in hemagglutination inhibition assay was significantly higher among CMV seronegative younger participants compared with CMV seropositive younger participants (385 vs. 142, P = 0.013). In contrast, among the older group, CMV serostatus was not associated with differential antibody titers (53 vs. 63, P = 0.75). These data suggest that CMV may shape immune response to neoantigens among younger persons; these groups should be included in future studies of immunosenescence and CMV. J. Med. Virol. 85:1557-1560, 2013. © 2013 Wiley Periodicals, Inc.
    Journal of Medical Virology 09/2013; 85(9):1557-60. · 2.37 Impact Factor
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    ABSTRACT: Additional drugs are needed for the treatment of cytomegalovirus (CMV) infection. Artesunate is an antimalarial drug that has activity against CMV in vitro and in a rodent model. Only a small number of case reports are available describing the clinical effects of artesunate on CMV infection, and these yielded inconsistent results. To evaluate the effect of artesunate on CMV infection, using blood samples collected from children who participated in malaria treatment trials. Quantitative CMV DNA PCR was performed on dried blood spots collected from 494 Ugandan children, who were randomized either to artesunate plus amodiaquine or sulfadoxine-pyrimethamine plus amodiaquine for acute malaria infection. Poisson regression was used to compare treatment regimens with respect to the change in the frequency and quantity of CMV detected that occurred before and after treatment. CMV was detected in 11.4% of children immediately prior to treatment and 10.7% 3 days later (p=0.70). The average quantity of CMV was 0.30 log10 copies per million cells higher on day 3 than at treatment initiation (95% CI 0.01-0.58, p=0.041). There was no measurable difference in either the frequency or quantity of CMV detected in blood between children randomized to the two treatment arms. A standard 3-day artesunate-containing antimalarial regimen had no detectable effect on CMV viremia in children with malaria. Longer treatment courses and/or higher doses of artesunate than those routinely used for malaria may be required for effective treatment of CMV infection.
    Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 07/2013; · 3.12 Impact Factor
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    ABSTRACT: BACKGROUND: Respiratory syncytial virus (RSV) is the most important cause of serious respiratory infections in young children. No prior studies using molecular techniques to examine RSV transmission in the community childcare setting have been performed. OBJECTIVES: We seek to characterize the molecular epidemiology of RSV transmission in childcare to evaluate the impact of RSV disease in a community-based population. METHODS: We sequenced RSV-positive nasopharyngeal samples from a prospective longitudinal study of respiratory illnesses among children enrolled in childcare during three winter seasons. Phylogenetic analysis was performed to identify unique viral strains. RESULTS: RSV was detected in 59 (11%) illnesses. Compared to RSV-negative illnesses, RSV-positive illnesses were associated with longer symptom duration and increased frequency of health care visits. Another respiratory virus was detected in 42 (71%) RSV-positive illnesses. RSV viral load did not differ between RSV-positive illnesses with and without another respiratory virus identified (P=0.38). In two childcare rooms, 50% of the children had RSV detected within six days of the first case. Five (38%) of 13 illness episodes from one childcare room were sequenced and shown to be the same viral strain, suggesting rapid child-to-child transmission within the room over a 16 day period. CONCLUSIONS: RSV is rapidly transmitted within childcare. Childcare facilities may serve as ideal sites for evaluation of new prevention strategies given the high burden of RSV disease in this population and the rapidity of RSV spread between children.
    Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 05/2013; · 3.12 Impact Factor
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    ABSTRACT: An assay to accurately quantitate CMV load in finger-stick collected dried blood spots (DBS) could potentially be useful for field studies or for analyzing patient self-collected specimens. We therefore assessed CMV DNA load in paired venipuncture-collected plasma samples and finger-stick DBS, using a previously validated quantitative polymerase chain reaction (PCR) assay. Assay variability, sensitivity, and changes in viral load during antiviral therapy in finger-stick DBS were compared to the reference plasma quantitative PCR assay, using 106 prospectively collected pairs of finger-stick DBS and plasma samples from 35 solid-organ transplant (SOT) patients. The DBS assay showed good agreement with the reference plasma viral load assay on the log10 scale (Pearson correlation coefficient 0.92, p < 0.001). The 95% limit of detection of the DBS assay was estimated at 2700 plasma copies/mL (675 plasma IU/mL). In 94% (76/81) of paired DBS and plasma samples above the limit of detection, the difference in CMV load was < 1 log10. CMV viral load changes during antiviral treatment were comparable in plasma and DBS. We conclude that finger-stick DBS provides a convenient sample type for quantitation of CMV load that correlates well with plasma levels. Future studies to optimize and evaluate this methodology for patient self-collected samples are warranted.
    Journal of clinical microbiology 05/2013; · 4.16 Impact Factor
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    ABSTRACT: BACKGROUND:: Standard doses of HSV suppressive therapy reduce plasma HIV-1 RNA levels (0.25-0.53 log10 copies/mL) among HIV-1/HSV-2 co-infected persons. Postulated mechanisms for this effect include direct inhibition of HIV-1 by acyclovir or indirect reduction by decreasing HSV-associated inflammation. We hypothesized that high-dose valacyclovir would further reduce plasma HIV-1 RNA, and that the effect would be mediated by greater suppression of HSV shedding. METHODS:: 34 participants with HIV-1 and HSV-2 not on antiretroviral therapy were enrolled into a randomized, open-label cross-over trial of valacyclovir 1000 mg twice daily or acyclovir 400 mg twice daily for 12 weeks, followed by a two week washout, and then the alternate treatment arm for 12 weeks. HSV DNA was measured from daily self-collected genital swabs for the initial 4 weeks of each arm and HIV-1 RNA was quantified from weekly plasma samples. RESULTS:: 28 participants provided plasma samples and genital swabs on both acyclovir and valacyclovir. The genital HSV-2 shedding rate was the same on valacyclovir and acyclovir (7.8% vs. 8.2% of days; RR, 0.95; 95% CI, 0.66-1.37; p=0.78). Plasma HIV-1 RNA was 0.27 log10 copies/mL lower on valacyclovir compared with acyclovir (95% CI, -0.41 to -0.14 log10 copies/mL; p<0.001); this was unchanged after adjustment for genital HSV-2 shedding. CONCLUSIONS:: High-dose valacyclovir reduces plasma HIV-1 RNA levels more than standard-dose acyclovir in HIV-1/HSV-2 seropositive persons not receiving antiretroviral therapy. The incremental reduction in plasma HIV-1 RNA achieved is not mediated by greater genital HSV-2 suppression.
    JAIDS Journal of Acquired Immune Deficiency Syndromes 03/2013; · 4.65 Impact Factor

Publication Stats

2k Citations
540.27 Total Impact Points

Institutions

  • 2007–2014
    • University of Washington Seattle
      • • Department of Laboratory Medicine
      • • Department of Medicine
      Seattle, Washington, United States
    • Fred Hutchinson Cancer Research Center
      • Division of Vaccine and Infectious Disease
      Seattle, Washington, United States
  • 2013
    • Seattle Children's Hospital
      Seattle, Washington, United States
  • 2012
    • Virginia Mason Medical Center
      Seattle, Washington, United States
  • 2009–2010
    • Seattle Children’s Research Institute
      Seattle, Washington, United States
  • 2007–2009
    • Dartmouth–Hitchcock Medical Center
      Lebanon, New Hampshire, United States