Rhoda Ashley Morrow

Acyclovir and transmission of HIV-1 from persons infected with HIV-1 and HSV-2.

BACKGROUND Most persons who are infected with human immunodeficiency virus type 1 (HIV-1) are also infected with herpes simplex virus type 2 (HSV-2), which is frequently reactivated and is associated with increased plasma and genital levels of HIV-1. Therapy to suppress HSV-2 reduces the frequency of reactivation of HSV-2 as well as HIV-1 levels, suggesting that suppression of HSV-2 may reduce the risk of transmission of HIV-1. METHODS We conducted a randomized, placebo-controlled trial of suppressive therapy for HSV-2 (acyclovir at a dose of 400 mg orally twice daily) in couples in which only one of the partners was seropositive for HIV-1 (CD4 count, > or = 250 cells per cubic millimeter) and that partner was also infected with HSV-2 and was not taking antiretroviral therapy at the time of enrollment. The primary end point was transmission of HIV-1 to the partner who was not initially infected with HIV-1; linkage of transmissions was assessed by means of genetic sequencing of viruses. RESULTS A total of 3408 couples were enrolled at 14 sites in Africa. Of the partners who were infected with HIV-1, 68% were women, and the baseline median CD4 count was 462 cells per cubic millimeter. Of 132 HIV-1 seroconversions that occurred after randomization (an incidence of 2.7 per 100 person-years), 84 were linked within couples by viral sequencing: 41 in the acyclovir group and 43 in the placebo group (hazard ratio with acyclovir, 0.92, 95% confidence interval [CI], 0.60 to 1.41; P=0.69). Suppression with acyclovir reduced the mean plasma concentration of HIV-1 by 0.25 log(10) copies per milliliter (95% CI, 0.22 to 0.29; P<0.001) and the occurrence of HSV-2-positive genital ulcers by 73% (risk ratio, 0.27; 95% CI, 0.20 to 0.36; P<0.001). A total of 92% of the partners infected with HIV-1 and 84% of the partners not infected with HIV-1 remained in the study for 24 months. The level of adherence to the dispensed study drug was 96%. No serious adverse events related to acyclovir were observed. CONCLUSIONS Daily acyclovir therapy did not reduce the risk of transmission of HIV-1, despite a reduction in plasma HIV-1 RNA of 0.25 log(10) copies per milliliter and a 73% reduction in the occurrence of genital ulcers due to HSV-2. (ClinicalTrials.gov number, NCT00194519.)

Efficacy Results of a Trial of a Herpes Simplex Vaccine

BACKGROUND Two previous studies of a herpes simplex virus type 2 (HSV-2) subunit vaccine containing glycoprotein D in HSV-discordant couples revealed 73% and 74% efficacy against genital disease in women who were negative for both HSV type 1 (HSV-1) and HSV-2 antibodies. Efficacy was not observed in men or HSV-1 seropositive women. METHODS We conducted a randomized, double-blind efficacy field trial involving 8323 women 18 to 30 years of age who were negative for antibodies to HSV-1 and HSV-2. At months 0, 1, and 6, some subjects received the investigational vaccine, consisting of 20 μg of glycoprotein D from HSV-2 with alum and 3-O-deacylated monophosphoryl lipid A as an adjuvant; control subjects received the hepatitis A vaccine, at a dose of 720 enzyme-linked immunosorbent assay (ELISA) units. The primary end point was occurrence of genital herpes disease due to either HSV-1 or HSV-2 from month 2 (1 month after dose 2) through month 20. RESULTS The HSV vaccine was associated with an increased risk of local reactions as compared with the control vaccine, and it elicited ELISA and neutralizing antibodies to HSV-2. Overall, the vaccine was not efficacious; vaccine efficacy was 20% (95% confidence interval [CI], -29 to 50) against genital herpes disease. However, efficacy against HSV-1 genital disease was 58% (95% CI, 12 to 80). Vaccine efficacy against HSV-1 infection (with or without disease) was 35% (95% CI, 13 to 52), but efficacy against HSV-2 infection was not observed (-8%; 95% CI, -59 to 26). CONCLUSIONS In a study population that was representative of the general population of HSV-1- and HSV-2-seronegative women, the investigational vaccine was effective in preventing HSV-1 genital disease and infection but not in preventing HSV-2 disease or infection. (Funded by the National Institute of Allergy and Infectious Diseases and GlaxoSmithKline; ClinicalTrials.gov number, NCT00057330.).

Comparison of Real-Time PCR Assays with Fluorescent-Antibody Assays for Diagnosis of Respiratory Virus Infections in Children

ABSTRACT Conventional fluorescent-antibody (FA) methods were compared to real-time PCR assays for detection of respiratory syncytial virus (RSV), influenza virus type A (FluA), parainfluenza virus types 1, 2, and 3 (PIV1, PIV2, and PIV3), human metapneumovirus (MPV), and adenovirus (AdV) in 1,138 specimens from children with respiratory illnesses collected over a 1-year period. At least one virus was detected in 436 (38.3%) specimens by FA and in 608 (53.4%) specimens by PCR (P < 0.001). Specimen quality was inadequate for FA in 52 (4.6%) specimens; 13 of these (25%) were positive by PCR. In contrast, 18 (1.6%) specimens could not be analyzed by PCR; 1 of these was positive by FA. The number of specimens positive only by PCR among specimens positive by PCR and/or FA was 18 (7.0%) of 257 for RSV, 18 (13.4%) of 134 for FluA, 25 (64.1%) of 39 for PIV1, 8 (88.9%) of 9 for PIV2, 17 (30.1%) of 55 for PIV3, and 101 (76.5%) of 132 for AdV. MPV was detected in 6.6% of all specimens and in 9.5% of the 702 specimens negative by FA. The mean number of virus copies per milliliter in specimens positive by both PCR and FA was significantly higher, at 6.7 × 107, than that in specimens positive only by PCR, at 4.1 × 104 (P < 0.001). The PCR assays were significantly more sensitive than FA assays for detecting respiratory viruses, especially parainfluenza virus and adenovirus. Use of real-time PCR to identify viral respiratory pathogens in children will lead to improved diagnosis of respiratory illness.

Brief Communication: Fatal Human Metapneumovirus Infection in Stem-Cell Transplant Recipients

Context Human metapneumovirus (hMPV) is a newly discovered respiratory virus. Contribution This study from a cancer referral center retrospectively examined bronchoalveolar lavage samples collected from 1995 to 1999 from hematopoietic cell transplant recipients with new or changing pulmonary infiltrates. The authors cultured hMPV in 5 of 163 symptomatic patients who were thought to have had the idiopathic pneumonia syndrome. Clinical findings included fever, cough, nasal congestion, respiratory failure, pulmonary hemorrhage, and culture-negative sepsis. Four of the patients died. Cautions All patients had moderate to severe symptoms. Implications Human metapneumovirus may cause respiratory failure in immunocompromised adults. The Editors Human metapneumovirus (hMPV) is a newly discovered respiratory virus associated with 10% to 25% of respiratory tract infections in young children (1, 2). This virus has been detected in respiratory disease at all ages (3, 4), although lower respiratory tract disease occurs more frequently in very young and very old persons and in those with underlying conditions. Clinical findings seen with hMPV infection are similar to those of respiratory syncytial virus (RSV) infection (5). As with RSV, serious disease associated with hMPV infection has been reported in immunocompromised patients. Deaths have been reported in a child (5), adults with hematologic malignant conditions (6), and adults after bone marrow or organ transplantation (7, 8). Pneumonia remains one of the most frequent serious complications of hematopoietic stem-cell transplantation (HCT) (9). Despite increased testing for pathogens in the transplantation setting, nearly 10% of myeloablative transplant recipients still develop the idiopathic pneumonia syndrome (10). We retrospectively evaluated archived bronchoalveolar lavage (BAL) fluid from patients undergoing HCT by using a sensitive hMPV-specific, real-time quantitative reverse transcriptase polymerase chain reaction (RT-PCR) assay to determine whether hMPV may be a cause of pulmonary morbidity and mortality in this setting. Methods Patients undergoing HCT who underwent BAL between 1995 to 1999 for evaluation of respiratory disease were eligible for inclusion if they had previously consented to diagnostic studies on their tissue and if residual specimens were available and were in good condition. We retrospectively reviewed clinical and laboratory records, biopsy specimens, and autopsy specimens of all patients with detected hMPV. We also evaluated stored BAL samples in 19 HCT recipients who were enrolled in a separate prospective study evaluating BAL after transplantation. Specimens Our institution has used a diagnostic algorithm for defining the cause of pneumonia for patients with new or changing pulmonary infiltrates since 1994 (11). We processed BAL samples in a centralized laboratory for pathogens, as previously described (11). Biopsy and autopsy specimens were submitted for routine bacterial, fungal, and acid-fast bacilli culture tests; underwent cytologic examination; and were tested for mycobacteria, Legionella species, Pneumocystis jiroveci, and fungal species. Additional studies performed in real time included culture and antibody staining of BAL fluid for cytomegalovirus (CMV), RSV, parainfluenza, and influenza viruses. For our study, we assayed BAL from patients with hMPV by using PCR for RSV (12), hMPV (13), parainfluenza types 1 and 3, influenza A and B, adenovirus, human herpesvirus-6 (14), and Aspergillus species (15). In addition, we performed broad-range fungal 18S rDNA PCRs to detect other fungal pathogens (16) and a -globin PCR (17) to determine whether amplifiable DNA was present in BAL fluid. The RT-PCR assay used probes that were designed to amplify 69 and 74 base-pair fragments of the fusion protein genes from the 2 major hMPV groups (13). This assay was sensitive to 10 copies per reaction and was specific for hMPV, as previously described (18). We also examined lung tissue from autopsy specimens of patients with known RSV, CMV, and hMPV infection near the time of death after tissue digestion by using the same RT-PCR methods. Paraffin-embedded tissues were examined for the presence of hMPV antigen by immunohistochemistry in the laboratory of Dr. Thijs Kuiken, Erasmus University, Rotterdam, the Netherlands (19). We included lung sections of an experimentally infected cynomolgus monkey with hMPV as positive controls. Role of the Funding Sources The National Institutes of Health, the Fred Hutchinson Cancer Research Center, and the Adult Leukemia Research Center provided funding for this study. The funding sources had no role in the design, analysis, interpretation, or writing of this manuscript or in the decision to submit the manuscript for publication. Results Patients To determine the presence of hMPV, we evaluated BAL specimens obtained throughout the calendar year from 163 symptomatic and 19 asymptomatic HCT recipients. The mean age of patients undergoing BAL was 42 years; only 10 patients (7%) were younger than 20 years of age. Most patients (99 of 163 patients [60%]) with BAL specimens had received allogenic bone marrow transplants from unrelated donors (57 of 163 patients [35%]), matchedrelated donors (24 of 163 patients [15%]), or mismatchedrelated donors (15 of 163 patients [9%]). Another 64 patients received peripheral blood stem-cell transplants (64 of 163 patients [39%]) from matched allogenic donors (16%), autologous donors (16%), or unrelated donors (4%). One patient received an unrelated cord blood stem-cell transplant. We detected hMPV in the tested BAL specimens of 5 of 163 patients (3.0%) and in 8 of 211 specimens (3.8%). Patients were between 24 and 54 years of age, and they had received different preparative regimens and sources of stem cells (Table). We detected hMPV in samples obtained between January and April. Clinical signs and symptoms around disease onset included a low-grade fever in 4 patients, cough in 3 patients, nasal congestion in 3 patients, and sore throat in 4 patients (although transplantation-related mucositis may have confounded some symptoms). Three patients had wheezing at diagnosis. We did not note sinusitis, otalgia, otitis media, conjunctivitis, or skin rashes. All patients eventually developed radiographic evidence of diffuse pulmonary disease, and we noted rapid progression of pulmonary infiltrates in 4 of 5 patients (Figure, top). Table. Characteristics of Hematopoietic Stem-Cell Transplant Recipients with Human Metapneumovirus Infection Figure. Computed tomography scan of chest in a patient with fatal human metapneumovirus infection after transplantation ( top ) and lung histologic study in a patient with human metapneumovirus pneumonia ( bottom ). Top. Bottom. inset arrow arrow All 5 cases of hMPV lower respiratory tract disease occurred early (<40 days) after transplantation. Lymphopenia was common, although 3 patients had neutrophil engraftment at the time of hMPV detection. Diffuse alveolar hemorrhage was present in 3 patients, and prominent alveolar macrophages were present in another patient. The quantity of hMPV ranged from 2104 RNA copies/mL to 3108 RNA copies/mL of BAL fluid (Table). Detection of Potential Co-Pathogens All BAL sample test results for patients with hMPV were negative for CMV, RSV, influenza viruses, adenovirus, and fungi by real-time PCR. We did not identify any accompanying pathologic or microbiological finding in 3 patients. Herpes simplex virus was isolated at autopsy in case 3, and parainfluenza was detected by culture and PCR in case 4. Case 2 had CMV isolated from lung tissue at autopsy only. We also tested BAL samples from patients with hMPV for human herpesvirus-6. Four of eight sample test results were negative, and the remaining 3 samples had low copies of human herpesvirus-6 DNA (53, 71, and 93 human herpesvirus-6 DNA copies/mL), which are levels compatible with detection of virus in latently infected cells (14). Treatment and Outcome We treated 3 of 5 patients who received a diagnosis of diffuse alveolar hemorrhage (cases 1, 4, and 5) with high-dose corticosteroids without discernable benefit. The hMPV viral load in case 4 decreased despite the use of corticosteroids (Table). Case 2 received aerosolized ribavirin for parainfluenza pneumonia, but additional respiratory samples were unavailable for testing. Progression of respiratory symptoms was rapid, with a median of 4 days (range, 1 day to 32 days) between initiation of oxygen therapy and death. Four patients died of respiratory complications at a median of 16 days (range, 1 day to 35 days) after the BAL was found to be positive for hMPV. Three patients (cases 1, 2, and 3) also had a culture-negative sepsis syndrome that required pressor agents. Clinical diagnoses at the time of death included idiopathic pneumonia in 3 patients; sepsis, shock, or both in 3 patients; and parainfluenza pneumonia in 1 patient. The mortality rate within 3 months of the first BAL in patients with hMPV was significantly higher than that in patients in whom hMPV was not detected (Wilcoxon rank-sum test; P= 0.014). We performed a limited postmortem examination of the lungs for case 2. We noted several bilateral pleural adhesions with diffuse alveolar damage and moderate hemorrhage. The cytologic changes suggested viral infection (Figure, bottom). Postmortem lung culture grew CMV that had not been previously detected. One of 2 archived frozen tissue samples, obtained at autopsy 8 days after bronchoscopy, was found to be positive for hMPV by RT-PCR but was negative by immunohistochemistry. One HCT recipient survived hMPV infection early in her transplantation course. This young patient had neutrophil engraftment when respiratory symptoms were noted. Nonetheless, she experienced a rapidly deteriorating respiratory course and, although she was never intubated, required supplemental oxygen for 2 months. Her hoarseness, dyspnea, and cou

Human Immunodeficiency Virus Acquisition Associated with Genital Ulcer Disease and Herpes Simplex Virus Type 2 Infection: A Nested Case-Control Study in Rakai, Uganda

To assess the timing of symptomatic genital ulcer disease (GUD) relative to human immunodeficiency virus (HIV) seroconversion, we studied 248 case subjects who underwent HIV seroconversion and 496 HIV-negative control subjects, at 3 interview visits conducted at 10-month intervals: visit 1, before HIV acquisition; visit 2, after seroconversion; and visit 3, 10 months after detection of seroconversion. Odds ratios (ORs) and 95% confidence intervals (CIs), for HIV acquisition, were estimated by logistic regression. HIV load was measured by RNA-polymerase chain reaction, and herpes simplex virus type 2 (HSV-2) serologic testing used HerpeSelect EIA with Western blot confirmation. The OR of HSV-2 seropositivity associated with HIV acquisition was 1.7 (95% CI, 1.2-2.4). Prevalence of GUD was increased among case subjects, at visits 2 (OR, 3.2; 95% CI, 1.9-5.3) and 3 (OR, 2.1; 95% CI, 1.1-3.9). HIV load was increased in HSV-2-seropositive case subjects, compared with that in HSV-2-seronegative subjects, at 5 (P=.04) and 15 (P=.02) months after seroconversion. HIV acquisition is associated with HSV-2 seropositivity, and GUD is increased after seroconversion. HIV load is increased in HSV-2-positive subjects who seroconverted, suggesting a role for treatment of HSV-2 infection in HSV-2-seropositive, dually infected individuals.

Clinical Disease in Children Associated With Newly Described Coronavirus Subtypes

OBJECTIVES. Coronaviruses cause upper respiratory illness and occasionally lower tract disease in susceptible populations. In this study we examined the prevalence of 4 human coronaviruses, including subtypes OC43, 229E, and the recently described NL63 and HKU1 in a pediatric population presenting to a childrens hospital. PATIENTS AND METHODS. Specimens collected over a 1-year period from pediatric patients presenting with acute respiratory illness were analyzed for the presence of 4 coronavirus subtypes using consensus and subtype-specific real-time reverse-transcription polymerase chain reaction assays. The demographic and clinical characteristics associated with coronavirus infection were examined retrospectively. RESULTS. Coronaviruses were detected in 66 of 1043 children. Eight, 11, 19, and 28 specimens were positive for subtypes 229E, NL63, OC43, and HKU1, respectively. Coronaviruses were detected throughout the study period; all 4 of the subtypes were present simultaneously in December. The acute clinical features were similar across subtypes. Of 32 children infected with a coronavirus as the sole respiratory pathogen, 13 had lower respiratory tract disease. Children whose only detectable respiratory virus was a coronavirus were more likely to have underlying chronic disease than were children coinfected with another respiratory virus. CONCLUSIONS. Although 4 subtypes of coronavirus were detected, the recently discovered coronavirus subtypes NL63 and HKU1 accounted for the majority of coronaviruses detected in our cohort of mostly hospitalized children with respiratory symptoms. New subtypes likely represent a substantial portion of previously unexplained respiratory illnesses.

The relationship between condom use and herpes simplex virus acquisition

Context We need other means to reduce the risk for transmitting genital herpes (herpes simplex virus type 2 [HSV-2]). Are condoms effective? Content In a trial of an ineffective HSV-2 vaccine, 1843 participants were divided into 3 groups according to the frequency of condom use (for 0% to 25%, 25% to 75%, and >75% to 100% of sexual acts). Frequent condom users had fewer HSV-2 infections. Compared with participants in the next lowest category, participants in a category had a 26% lower risk for HSV-2 infection. Limitation In this observational cohort study, many unmeasured factors could also contribute to altered rates of HSV-2 acquisition. Conclusion Condom use is associated with a lower rate of acquisition of HSV-2. The Editors Genital herpes is a common sexually transmitted infection that can be transmitted during episodes of recurrent lesions and during subclinical shedding (1). In the absence of an effective vaccine, condoms have been routinely recommended for prevention of transmission, and a recent study showed that daily antiviral therapy also decreases the risk for transmission of herpes simplex virus type 2 (HSV-2) in discordant couples (2, 3). In a previous study of monogamous HSV-2discordant couples who were enrolled in an ineffective candidate HSV-2 vaccine trial, we showed that condoms protect women from HSV-2 infection (4). However, very few cases of genital HSV-2 occurred among men who were sexual partners of women infected with HSV-2, precluding definitive conclusions about the effectiveness of condoms for prevention of transmission to men. We present data from a concurrent trial of the candidate vaccine among HSV-2seronegative persons attending sexually transmitted disease clinics (5). A total of 1862 participants were enrolled in this study; 85 cases of genital herpes were documented in men, and 33 cases were documented in women. We analyzed the effect of condom use on HSV acquisition in this prospectively followed cohort of men and women. Methods Study Sample Participants included in this analysis took part in a randomized, double-blind, placebo-controlled efficacy trial of a candidate subunit HSV-2 vaccine that was subsequently shown to be ineffective (5). The trial involved 22 centers located at sexually transmitted disease clinics and enrolled 1862 participants. Initial serologic testing was done at screening; participants who were seronegative for HIV and HSV-2 and reported 4 or more sexual partners in the past year or 1 or more sexually transmitted diseases in the past year were eligible to enroll. The effectiveness of condom use among the 528 discordant couples enrolled in a parallel vaccine study was reported previously (4, 5). Participants were enrolled and followed for 18 months, during which they were evaluated at 11 study visits. At enrollment, we collected demographic information and information about sexual history. At each study visit, we took blood samples and recorded the following information about sexual history, which described behavior since the last visit: frequency of sexual activities, defined as vaginal or anal intercourse; frequency of condom use; number of partners; number of new partners; and number of partners with a known history of genital herpes. The information regarding number of partners was gender-specific. In addition, participants were counseled routinely about safer sexual behavior and were offered condoms at each study visit. Genitourinary signs and symptoms were evaluated as needed at additional interim visits. Laboratory Methods The Western blot assay done at the University of Washington, Seattle, Washington, established HSV serologic status at study entry and was used to document seroconversion (6). Type-specific cultures using standard techniques were done at local study sites. Statistical Analysis Acquisition of HSV-2 was defined by seroconversion on the Western blot assay or by a positive culture for HSV-2. Time to HSV-2 acquisition was defined as the number of days from screening to the first positive culture for HSV-2 or as the midpoint between the last negative result of the HSV-2 antibody test and the first positive result of the HSV-2 antibody test. In this analysis of condom use and HSV acquisition, we included the time from screening to enrollment in the study, whereas in the vaccine trial participants were followed beginning at enrollment. Thus, our report includes 109 participants who were not included in the efficacy analysis of the original vaccine trial. Twenty of these participants seroconverted to HSV-2 during the screening period before enrollment, and 89 were lost to follow-up after enrollment. Participants who did not acquire HSV-2 were censored at the last blood draw taken during the study or at enrollment if they did not report any sexual activity thereafter. Participants who reported no sexual activity for the entire time from screening to study termination were excluded from the analysis because they were not at risk for HSV-2 infection. Participants with follow-up longer than 65 days beyond the 18 months specified in the protocol (3%) were censored at day 605. Participants who were seronegative for HSV type 1 (HSV-1) and HSV-2 at screening were included in the analysis of HSV-1. Time to HSV-1 acquisition was defined as the number of days from screening to the first positive culture for HSV-1 or the midpoint between the last negative result of the HSV-1 antibody test and the first positive result of the HSV-1 antibody test. Participants who did not acquire HSV-1 were censored at the last blood draw or at enrollment if they did not report any sexual activity thereafter. Participants who reported no sexual activity for the duration of the study were excluded from the analysis of HSV-1. KaplanMeier curves, log-rank tests, and univariate and multivariate Cox regression models were used to determine baseline risk factors associated with HSV-2 acquisition. To relate sexual behavior to HSV-2 acquisition during the study, we constructed time-dependent covariate Cox regression models. The analysis time was divided into four 150-day intervals, and information about sexual history collected at interim visits was used to calculate covariate summaries for each period. Because continuous variables did not satisfy the assumption of a linear effect in the log hazard, they were categorized. Our choice for the cut-points was motivated by maintaining equal numbers of participants in each category (for example, age was split at the median value, 27 years), by consistency with observed risk patterns, or by interpretation considerations. Frequency of sexual activity was expressed as the average number of sexual acts per week in the time period, calculated by averaging the reported estimates over the visits for each interval. This average was then categorized as greater than 2 versus 2 or fewer to correspond to observed risk patterns. Use of condoms during the study period was described categorically in each interval (used for 0% to 25%, for 25% to 75%, or for >75% of sexual acts). This grouped linear parameterization was chosen to remain consistent with published analyses (4) while allowing a doseresponse relationship, assuming constant change in the risk with increasing category of condom use. The use of condoms was not evaluated during intervals for which the participants did not report any sexual activity. The number of partners reported was summarized for each period and was modeled in a binary fashion. Partner cut-points were chosen for interpretation reasons to describe ways in which this patient group may differ from monogamous couples who were studied in a previously published report addressing condom use and infection with HSV (4). Total number of partners was modeled as more than 1 versus 1 or fewer, and both new partners and partners with a history of genital herpes were modeled as any versus none. These analyses did not adjust for receipt of placebo versus receipt of vaccine, because this factor was not statistically significant in acquisition of HSV and did not influence the covariates of interest for this study. An interaction term between condom use and gender was used to check the hypothesis of a difference in the effect of condoms by gender and to provide gender-specific estimates of condom use. Two-sided P values for model covariates were calculated by using the likelihood ratio test. The same methods were used to explore baseline risk factors and time-varying risk factors for time to infection with HSV-1. Poisson regression was used to provide P values for comparisons involving incidence rates. Tests for changes in sexual behavior with time used generalized estimating equations. Statistical analyses were done by using Stata statistical software (version 8.1, Stata Corp., College Station, Texas). Role of the Funding Source The funding for the analyses for this study was provided by federal grants; design, data analysis, and interpretation were done at the University of Washington. The initial clinical trial was funded by Chiron Corporation. This study was supported in part by National Institutes of Health Herpes Program Project Grant AI-30731 and Centers for Disease Control and Prevention Research Initiative UR6/CCU017828-02. Results Of the 1862 participants who enrolled for the vaccine trial, 19 did not report any sexual activity during the entire study and thus were excluded from this analysis. The remaining 1843 participants included 1365 men and 478 women. The median age of the participants was 27 years. Sixty-two percent were white, 32% were African American, and 6% were people of other races; 1184 participants (64%) were seropositive for HSV-1 at study entry. Most men and women qualified for the study by reporting 4 or more partners in the past year (66% of men, 70% of women); some reported 1 or more sexually transmitted diseases in the past year (12% of men, 19% of women); and the remainder met both criteria (22% of men, 1

Evaluation of quantitative and type-specific real-time RT-PCR assays for detection of respiratory syncytial virus in respiratory specimens from children.

Abstract Background: Respiratory syncytial virus (RSV) is a major cause of lower respiratory tract morbidity in young children and immunosuppressed patients. Objectives: To rapidly and accurately quantify and subtype RSV in respiratory samples, we developed and evaluated two real-time RT-PCR assays. Study design: A quantitative assay was designed using primers for a consensus region of the matrix protein gene and a subtype-specific assay for RSV-A and RSV-B detection was designed using primers for the polymerase gene. Quantitative RSV RT-PCR results of pediatric nasal wash samples submitted to the University of Washington Virology Laboratory from December 2002, through May 2003, were compared to those of an indirect fluorescent antibody RSV antigen detection assay (FA). Results: Specificity of the RT-PCR assay was high, with no amplification of eleven common respiratory viruses and eight herpes viruses. Among 751 samples, RSV was detected in 267 (35.6%) by FA and in 286 (38.1%) by RT-PCR. Median RSV copy number in nasal wash samples that were positive by both FA and RT-PCR was 2.5×107 copies/mL versus a median of 3.0×104 copies/mL for samples positive by RT-PCR only (P<0.001). The detection and quantity of RSV in respiratory specimens was associated with younger age, but not with gender or hospitalization. Among positive samples from this Seattle cohort, 52% were subtype A and 48% were subtype B. Both subtypes were detected with similar viral loads among all patient groups (stratified by age, gender, and hospitalization), and throughout the specimen collection period. Conclusions: These real-time RT-PCR assays provide a rapid, specific, and highly sensitive alternative for detecting, quantifying, and subtyping RSV in clinical specimens.

Knowledge of Partners’ Genital Herpes Protects against Herpes Simplex Virus Type 2 Acquisition

BACKGROUND Prospective studies of herpes simplex virus type 2 (HSV-2) infection in discordant couples have shown a low rate of transmission. However, unlike partners with genital herpes in prospectively monitored couples, most persons who transmit genital herpes are not aware of having the infection. METHODS Because HSV has a short incubation period and most persons who acquire genital herpes can identify the transmitting partner, a time-to-event design was used to assess risks of HSV acquisition among patients with newly acquired genital herpes. RESULTS Among 199 persons with laboratory-documented newly acquired genital herpes, the median duration of the sexual relationship with the transmitting partner was 3.5 months, and the median number of sex acts before transmission was 40. The median time to HSV-2 acquisition was greater among participants whose partners disclosed that they had genital herpes, compared with participants whose partners did not disclose their status (270 vs. 60 days; P = .03). In multivariate models, having a partner who disclosed that he or she had genital herpes remained a strong protective factor against genital HSV-2 acquisition (hazard ratio, 0.48 [95% confidence interval, 0.25-0.91]). CONCLUSION These findings suggest that testing persons with HSV type-specific serologic assays and encouraging disclosure may result in a decreased risk of HSV-2 transmission to sex partners.

Detection and quantification of human metapneumovirus in pediatric specimens by real-time RT-PCR

Abstract Background: Human metapneumovirus (hMPV), a recently identified virus, causes respiratory illness in children. Objectives: A real-time reverse transcription-polymerase chain reaction (RT-PCR) assay was developed and used to detect and quantify hMPV in respiratory specimens. Study design: The quantitative RT-PCR assay amplified an approximately 70 base pair fragment from the hMPV fusion protein gene. The assay was validated and used to test respiratory specimens obtained from children seen at a hospital in Seattle, Washington, from December 2002 through May 2003. Results: The assay detected 1000hMPV copies/mL of specimen, did not detect 19 other respiratory viruses, and was able to detect and accurately quantify isolates from the four known hMPV genetic lineages in a proficiency panel of 20 previously tested samples. hMPV was detected in 52 (7.2%) of 719 pediatric respiratory specimens. The mean log10copies/mL of hMPV in the 52 positive specimens was 7.67 (range=4.59–10.60). Children aged 7–12 months had a significantly higher hMPV prevalence (12.4%) than did children younger than 7 months (4.7%) (P <0.005). Children in this age group also had significantly higher levels of hMPV in their respiratory specimens (mean log8.43 copies/mL) than did the younger children (mean log6.93copies/mL) (P =0.0025). Conclusions: The rapid real-time RT-PCR assay described here is a sensitive test for clarifying the epidemiology of and diseases associated with hMPV.