Herpes Simplex Virus Cultures Research Articles

A double-blind, randomized, placebo-controlled trial of acyclovir in late pregnancy for the reduction of herpes simplex virus shedding and cesarean delivery

Objective: The purpose of this study was to assess the efficacy of acyclovir in the reduction of herpes simplex virus culture and polymerase chain reaction positivity and cesarean delivery. Study Design: Women with recurrent genital herpes simplex virus were randomized to acyclovir 400 mg three times daily or placebo from 36 weeks of gestation until delivery. A subset of daily specimens for herpes simplex virus culture and DNA polymerase chain reaction was self-collected. Analyses used χ2, Fisher exact, and Mann-Whitney U tests. Results: Lesions occurred at delivery among 11 of 78 women (14%) who received placebo and 4 of 84 women (5%) who received acyclovir (P =.08). Herpes simplex virus culture and polymerase chain reaction positivity near delivery occurred in 7% and 34% women in the placebo group and 0 and 2% in the acyclovir group (P =.03 and <.01, respectively). Cesarean delivery for herpes simplex virus occurred in 8 of the women (10%) in the placebo group and in 3 of the women (4%) in the acyclovir group (P =.17). Despite reductions in herpes simplex virus detection, 6% of the women who received acyclovir had herpes simplex virus detected by polymerase chain reaction on >20% of days. Neonatal outcomes were similar between groups. Conclusion: Acyclovir significantly reduced, but did not eliminate, herpes simplex virus lesions and detection in late pregnancy. (Am J Obstet Gynecol 2003;188:836-43.)

Effect of serologic status and cesarean delivery on transmission rates of herpes simplex virus from mother to infant.

Neonatal herpes most commonly results from fetal exposure to infected maternal genital secretions at the time of delivery. The risk of transmission from mother to infant as it relates to maternal herpes simplex virus (HSV) serologic status and exposure to HSV in the maternal genital tract at the time of labor has not been quantified. Furthermore, no data exist on whether cesarean delivery, the standard of care for women with genital herpes lesions at the time of delivery, reduces HSV transmission. To determine the effects of viral shedding, maternal HSV serologic status, and delivery route on the risk of transmission of HSV from mother to infant. Prospective cohort of pregnant women enrolled between January 1982 and December 1999. A university medical center, a US Army medical center, and 5 community hospitals in Washington State. A total of 58 362 pregnant women, of whom 40 023 had HSV cultures obtained from the cervix and external genitalia and 31 663 had serum samples tested for HSV. Rates of neonatal HSV infection. Among the 202 women from whom HSV was isolated at the time of labor, 10 (5%) had neonates with HSV infection (odds ratio [OR], 346; 95% confidence interval [CI], 125-956 for neonatal herpes when HSV was isolated vs not isolated). Cesarean delivery significantly reduced the HSV transmission rate among women from whom HSV was isolated (1 [1.2%] of 85 cesarean vs 9 [7.7%] of 117 vaginal; OR, 0.14; 95% CI, 0.02-1.08; P =.047). Other risk factors for neonatal HSV included first-episode infection (OR, 33.1; 95% CI, 6.5-168), HSV isolation from the cervix (OR, 32.6; 95% CI, 4.1-260), HSV-1 vs HSV-2 isolation at the time of labor (OR, 16.5; 95% CI, 4.1-65), invasive monitoring (OR, 6.8; 95% CI, 1.4-32), delivery before 38 weeks (OR, 4.4; 95% CI, 1.2-16), and maternal age less than 21 years (OR, 4.1; 95% CI, 1.1-15). Neonatal HSV infection rates per 100 000 live births were 54 (95% CI, 19.8-118) among HSV-seronegative women, 26 (95% CI, 9.3-56) among women who were HSV-1-seropositive only, and 22 (95% CI, 4.4-64) among all HSV-2-seropositive women. Neonatal HSV infection rates can be reduced by preventing maternal acquisition of genital HSV-1 and HSV-2 infection near term. It can also be reduced by cesarean delivery and limiting the use of invasive monitors among women shedding HSV at the time of labor.

The prevalence of herpes simplex virus shedding and infection in the oral cavity of seropositive patients undergoing head and neck radiation therapy.

Herpes viruses are characterized by their ability to establish and maintain a latent infection that can reactivate. Only 2 preliminary studies have examined herpes simplex virus (HSV) reactivation in patients receiving head and neck radiotherapy. The role of radiation therapy in the reactivation of a latent virus has not been established. The purpose of the present study was to evaluate the incidence of HSV reactivation in patients receiving radiation treatment for head and neck malignancies. Twenty patients, 19 of whom were HSV seropositive, undergoing head and neck radiation therapy were assessed weekly before and during radiation therapy, and HSV cultures were completed during cancer treatment. Only 3.6% of the cultures were positive for HSV during radiation therapy. HSV was cultured in 4 men receiving a mean of 6,000 cGy to the head and neck area. Recovery from HSV was seen in patients nearing completion of radiation therapy. The results of this study suggest that HSV reactivation is not common during radiation therapy. Therefore, this study does not support prophylaxis of HSV in patients undergoing head and neck irradiation.

Longitudinal study of genital infection by herpes simplex virus type 1 in western Scotland over 15 years

Although herpes simplex virus type 2 (HSV-2) is regarded as causing most cases of genital herpes, preliminary reports suggest that the type 1 virus (HSV-1) is increasingly the cause of...

Detection and subtyping of Herpes simplex virus in clinical samples by LightCycler PCR, enzyme immunoassay and cell culture

BackgroundPrompt laboratory diagnosis of Herpes simplex virus (HSV) infection facilitates patient management and possible initiation of antiviral therapy. In our laboratory, which receives various specimen types for detection of HSV, we use enzyme immunoassay (EIA) for rapid detection and culture of this virus. The culture of HSV has traditionally been accepted as the diagnostic 'gold standard'. In this study, we compared the use of real time PCR (LightCycler) for amplification, detection and subtyping of specific DNA with our in-house developed rapid and culture tests for HSV.ResultsThe LightCycler PCR (LC-PCR) detected and subtyped HSV in 99% (66/67) of HSV positive specimens, compared to 81% (54/67) by rapid antigen EIA or 57% (36/63) by culture. A specimen was considered positive when two or more tests yielded HSV identifications or was culture positive. Discordant results were confirmed with an in-house developed PCR-ELISA or DNA sequence analysis. The typing results obtained with the LC-PCR and by culture amplified test were completely concordant.ConclusionsThis study showed that the LC-PCR provided a highly sensitive test for simultaneous detection and subtyping of HSV in a single reaction tube. In addition to increased sensitivity, the LightCycler PCR provided reduced turn-around-times (2 hours) when compared to enzyme immunoassay (4 hours) or culture (4 days).

Standardization of molecular amplification testing in the clinical microbiology laboratory

Where are we in the process of standardizing NAA testing in the clinical microbiology laboratory? In general, we are in the early stage with only minimal progress seen in HCV and HIV-1 testing. However, more technological advances in both the pre-analytical and analytical arenas may expedite this complex process. Specimen processing is a priority for some companies at the present time, with particular focus on automation and commercially available test reagents. Specimen processing is no doubt a roadblock in the path to standardization. Recently, we have seen the introduction of real-time PCR technology (TaqMan and LightCycler). This technology combines the amplification and detection steps, shortens the turnaround time, adds the option of quantitation (Figure 4), and essentially removes the contamination threat. In fact, the LightCycler even has been used to replace culture for Herpes simplex virus in one prominent clinical laboratory (10). This type of advancement in molecular testing further consolidates the entire process and brings the idea of NAA standardization even closer to reality.

First episodes of genital herpes in a Swedish STD population: a study of epidemiology and transmission by the use of herpes simplex virus (HSV) typing and specific serology

Objectives: To determine the proportion of herpes simplex virus type 1 (HSV-1) and HSV type 2 (HSV-2) in first episodes of genital herpes. To evaluate the use of HSV specific...

Herpes Simplex Virus DNA in Amniotic Fluid without Neonatal Infection

Twenty-one pregnant women were studied to determine the distribution of herpes simplex virus (HSV): 10 had symptomatic genital herpes, including 1 with primary cervical HSV infection, and 11 had asymptomatic genital herpes. Samples from vesicles, the cervix, and amniotic fluid (AF) were analyzed with 2 separate polymerase chain reaction (PCR) tests and with viral culture. For newborns, clinical examinations and pharyngeal HSV cultures were performed, and cord blood IgM antibodies to HSV were measured. HSV DNA was present in the AF of 3 women with symptomatic HSV infection, but all cultures were negative. HSV was detected more often with PCR than with culture, in both vesicle and cervical samples. For the asymptomatic group, all AF samples were negative, whereas 4 cervical samples were positive by PCR (none were positive by culture). All isolates were HSV type 2. All infants were healthy, and none had cord blood IgM antibodies to HSV, including those with PCR-positive AF.

Herpes simplex virus type 2 shedding in human immunodeficiency virus-negative men who have sex with men: frequency, patterns, and risk factors.

We investigated the frequency, site, and risk factors for herpes simplex virus (HSV) shedding in 30 human immunodeficiency virus (HIV)-negative HSV type 2 (HSV-2)-seropositive men who have sex with men. Subjects collected daily HSV culture samples from genital, perianal, and oral areas for 100 days and maintained diaries of signs and symptoms. Sixteen men (53.3%) shed HSV-2, and 9 (56.3%) of 16 men who were also HSV type 1 (HSV-1)-seropositive shed HSV-1. Overall, HSV-2 was isolated on 3.1% of the days; 68% of the isolations were on days that lesions did not occur. HSV-2 shedding was predominantly perianal (83.3%). HSV-1 was isolated on 2.1% of the days; 23 of 24 HSV-1 isolates were from oral areas. Rates of perianal or genital shedding were 6.6% on the days that participants reported prodromal symptoms and 1.9% on the days that participants did not report prodromal symptoms (P<.001). Men seropositive for both HSV-1 and HSV-2 were significantly more likely to shed HSV-2 (odds ratio, 4.1; 95% confidence interval, 1.4-11.9) than were HSV-2-seropositive men. HSV-2-seropositive men who have sex with men have frequent subclinical HSV-2 shedding, usually from the perianal area, and more frequent prodromal HSV-2 shedding.

Do antepartum herpes simplex virus cultures predict intrapartum shedding for pregnant women with recurrent disease?

OBJECTIVE: To examine antenatal screening as a predictor of intrapartum shedding of herpes simplex virus (HSV) and to determine its usefulness in guiding the appropriate route of delivery for patients with recurrent HSV in pregnancy. METHODS: A population of 198 pregnant women with a history of recurrent genital HSV were cultured in the last weeks of their pregnancy by specially-trained personnel and intrapartum by their delivering attendants. RESULTS: Of cultures from a total of 906 antenatal visits, 17% were culture positive, with an asymptomatic shedding rate of 3.4%. Asymptomatic shedding occurred in 12.6% of women. Over the 8-week antepartum period, viral culture-positivity rates for each visit ranged from 11% to 19.5%. This provided an expected delivery culture-positivity rate of 15.3%. However, actual intrapartum viral culture positivity occurred in only three of 191 women (1.5%; P < 0.001). Because previous studies have suggested antepartum culture positivity fails to predict intrapartum viral shedding, evaluations, including cultures, as well as predictive values for subsequent culture positivities, were determined under the supervision of an infectious disease specialist. Under these conditions, positive predictive values were 59% when the interval between visits was 2 days, but only 19% when days between visits were >2 (P < 0.0001). No cases of neonatal herpes were seen in this population, although cesarean deliveries were performed in 31% of the patient population, with genital herpes as the indication for 56% of those. CONCLUSIONS: Antepartum serial screening by viral culture is not predictive of an infant's risk of intrapartum viral exposure when conducted at weekly intervals. However, more frequent assessments of patients can be predictive of an infant's exposure risk to HSV; for patients with frequent recurrent disease near term or primary infection in pregnancy, frequent late antepartum screening may be appropriate.

Performance of six cell lines in the suspension-infection test used for the detection of herpes simplex virus

Six cell lines were assessed in the suspension-infection (SI) test for suitability for the rapid culture of herpes simplex virus (HSV). For the SI test, the specimen was combined in growth medium with trypsinized, suspended culture cells before allowing the cells to settle into a monolayer growth pattern. The cells tested in the SI assay were MV1-Lu, vero, C1008, BSC-1, RD, and MRC-9 cells. Fifty clinical specimens composed of 7 HSV-1–positive samples, 9 HSV-2–positive samples, 12 positive for HSV (not typed), and 22 HSV-negative samples were tested. For the detection of HSV in clinical specimens, MV1Lu cells were most sensitive and demonstrated large, darkly stained foci of virus infection.

Herpes simplex virus shedding among human immunodeficiency virus-negative men who have sex with men: site and frequency of shedding.

The frequency and anatomic location of subclinical and symptomatic herpes simplex virus (HSV) shedding were evaluated among human immunodeficiency virus (HIV)-negative HSV-2-seropositive men who have sex with men (MSM). Eight men attended a research clinic daily for 30 days for a detailed genital examination and anoscopy with colposcopy to detect herpes lesions. HSV cultures were obtained daily from four sites (perianal, urethral, penile shaft, and oral) at home and the research clinic. Signs and symptoms of genital herpes were recorded by the participants and clinician. Three (37.5%) of the men shed HSV. Overall, the 8 men shed HSV on 5.5% of days cultures were obtained and shed subclinically on 2.7% of days. All HSV shedding was perianal or rectal; only 1 symptomatic recurrence, concurrent with an external perianal lesion, was detected by anoscopy. Subclinical HSV shedding was frequent among HIV-negative MSM, and anoscopy with colposcopy did not increase the detection rate of rectal HSV lesions or shedding.

Occult herpes family viruses may increase mortality in critically ill surgical patients

Background: Effects of cytomegalovirus (CMV) or herpes simplex virus (HSV) infection on surgical intensive care unit (SICU) patients’ hospital outcome are unknown. METHODS: Between July 1, 1994, and September 1, 1995, general SICU patients with persistent sepsis and no bacterial or fungal source identifiable had viral cultures obtained. Patients with positive broncho-alveolar lavage, blood, skin, or sputum cultures for CMV or HSV were studied. RESULTS: Twenty eligible patients had positive viral cultures during the study period, and 85% of these patients developed subsequent bacterial and/or fungal infections. Mortality was significantly higher following viral infection than in chronic SICU patients (65% vs 35%, P <0.006). Patients with thrombocytopenia complicating their viral infection had significantly higher mortality than those without thrombocytopenia (92% vs 25%, P <0.004). CONCLUSIONS: At least 14% of critically ill surgical patients have occult infection or reactivation of herpes family viruses. These viruses have known immunosuppressive effects, which may predispose chronic SICU patients to subsequent bacterial and fungal infection, and subsequent organ system failure and death.

Rapid detection, culture-amplification and typing of herpes simplex viruses by enzyme immunoassay in clinical samples

Background: The laboratory diagnosis of herpes simplex infection may require rapid (direct) tests, as well as cell cultures, for detection of the virus in clinical samples. The quantity of virus present in clinical samples is variable and this may depend on the period from onset of rash. In addition, not all patients may show obvious symptoms with this infection. The successful culture of herpes simplex virus requires prompt transportation after collection of the specimen as the virus is easily inactived. Hence, rapid and culture tests would enable detection of non-viable and viable viruses. Study Design: We describe the rapid detection of HSV by EIA directly in various clinical samples using commercially available polyclonal sera. In addition, specimens were inoculated in microwell cell cultures and 4 days post inoculation the culture fluids were tested for HSV and subtyped by a similar EIA (culture amplified EIA). Results: The direct EIA showed an endpoint detection of 100 TCID 50/ml, sensitivity of 92% (all specimen types) and specificity of 100%. The direct EIA sensitivity was 97% in non-genital specimens and 88% in genital specimens. The culture amplified EIA showed a sensitivity of 95% compared to all confirmed HSV positive samples. Conclusions: The results of the HSV rapid tests were available within 24 h from receipt of specimens. Specimens which were culture negative/direct EIA positive were confirmed by blocking antisera. Culture positive specimens which were direct EIA negative were confirmed by subtyping of the virus.

Genital Ulcers as a Presenting Feature of Primary Epstein-Barr Virus Infection † 8

Genital ulcers (GU) have been rarely described in temporal association with acute Epstein-Barr Virus (EBV) infection, but a cause-and-effect relationship has not been established. A previously healthy 14-year-old girl with no history of sexual activity developed fever, sore throat, and low back pain; a day later, five painful ulcers appeared over her external genitalia. A hemogram was normal, herpes simplex virus (HSV) culture of the lesions was negative, and bacterial GU cultures grew normal flora. Her fever worsened(40.2°C) and the genital ulcers continued to enlarge. Examination on day 5 of GU showed deep, tender, hemorrhagic, coalesced ulcers involving both labia majora and minora, 2 mm aphthous ulcers on her buccal mucosa, no lymphadenopathy, and the hymen was estrogenized and without interruption. HSV cultures, ANA, VDRL, and HIV antibodies were negative. She was empirically treated with Augmentin®(2d) and then metronidazole (7d), as well as oral methylprednisolone (5d). On day 9 of GU, she was afebrile and the lesions were smaller and not hemorrhagic. A tentative diagnosis of Behcet syndrome was made. On day 15 of GU, she developed sore throat and dysphagia, but no fever. Examination revealed exudative tonsillitis and cervical lymphadenopathy; the ulcers were granulating and less tender. Her Monospot, EBV VCA IgG and IgM, and anti-EA (1:40) were positive; EBNA was negative. Whole blood, serum, and buffy coat obtained on day 16 of GU and the genital culture specimen obtained on day 5 (10 days before primary EBV infection was diagnosed) were assayed for EBV DNA using the polymerase chain reaction (PCR). The PCR assay was used to amplify a 177 bp region in the LMP-2A gene of EBV; the sensitivity of this assay was 8 EBV copies per 105 cells. All specimens were positive for EBV DNA. Conclusion: EBV DNA was found in genital ulcer specimens obtained 10 days before symptoms of EBV-related infectious mononucleosis were apparent. EBV should be considered as an etiologic agent of acute, non-venereal genital ulcers.

Oral disease in terminally ill cancer patients with xerostomia.

Xerostomia is common among patients with advanced cancer and is likely to contribute to oral disease. This study determined the prevalence of oral signs and symptoms among a group of 70 terminally ill cancer patients [25 male, 45 female; age range 42-88 (mean 66) years] complaining of oral dryness, and examined the associated oral microflora. Imprint cultures for yeasts, coliforms and staphylococci were collected from the tongue and, in denture wearers, from the plate and denture fitting surface. A swab was collected for culture of herpes simplex virus. 68 patients (97%) complained of oral dryness during the day and 59 patients (84%) complained of oral dryness at night. Oral soreness was reported by 22 patients (31%). 46 patients (66%) had difficulty talking and 36 (51%) reported difficulty eating. Of the 56 denture wearers, 40% complained of denture problems. On examination, 63 (90%) of the patients had clinically dry mouths. Oral mucosal abnormalities were detected in 45 patients (65%), most commonly erythema (20%), coated tongue (20%), atrophic glossitis (17%), angular cheilitis (11%) and pseudomembraneous candidosis (9%). 47 (67%) of the patients carried yeasts, 18 (26%) were carriers of Staphylococcus aureus and 13 (19%) carried coliforms. Herpes simplex virus was isolated from 5 patients, of whom 2 had herpetic stomatitis. Oral complications and abnormalities of the oral microflora can be detected among significant numbers of terminally ill cancer patients with xerostomia.

Herpes simplex virus-associated erythema multiforme in prepubertal children.

To examine clinical associations, evolution of the condition, and response to treatment of erythema multiforme (EM) in prepubertal children. A retrospective case series evaluation of children younger than 13 years with EM. Ambulatory care university hospital. Referral patients from pediatricians serving a population of 3.2 million. Results of treatment of each EM episode with topical acyclovir or oral acyclovir at a dose of 25 mg/kg per day and 6-month prophylaxis with oral acyclovir at a dose of 20 mg/kg per day were evaluated. Age at EM onset, preceding illness, and number and duration of episodes during a 3-year period were recorded. Twelve children (7 boys and 5 girls) in whom herpes simplex virus (HSV)-associated EM developed were evaluated. Preceding lesions were herpes labialis in 8 children and herpes facialis in 2 children. Two children had no obvious HSV lesion. The mean age at onset of disease was 8.1 years, and the mean time from the preceding HSV to the onset of skin lesions was 3.9 days (range, 0-11 days). Episodes of EM lasted a mean of 10.6 days. In 9 children, the EM was recurrent, with a mean of 2.6 episodes per year. All 12 children, including those with negative viral cultures for HSV or no HSV history had HSV detected in their target lesions by polymerase chain reaction amplification of DNA obtained from skin biopsy specimens. Six of 12 children were treated with oral acyclovir at a dose of 25 mg/kg per day for 1 or more individual episodes, without reduction in the episode. Three children underwent 6-month prophylaxis with oral acyclovir at a dose of 20 mg/kg per day and remained disease free during treatment. After discontinuation of the prophylactic treatment with acyclovir, 1 child relapsed at 4 months. The other 2 children had no further episodes during a 3-year period. The HSV-associated EM is a recurrent disease that can be precipitated by sun exposure and does not progress to Stevens-Johnson syndrome. Childhood HSV-associated EM may be unresponsive to treatment with oral steroids or oral or topical acyclovir. Frequent recurrences of EM may be abrogated by prophylactic treatment with acyclovir.

Herpes simplex encephalitis treated with acyclovir: diagnosis and long term outcome

OBJECTIVESThe frequency and characteristics of the long term sequelae of herpes simplex encephalitis were assessed after treatment with acyclovir.METHODSPatients were included if they were treated with acyclovir and the diagnosis...

Herpes simplex virus seropositivity and reactivation at delivery among pregnant women infected with human immunodeficiency virus-1

Objective: Our purpose was to determine whether pregnant women infected with human immunodeficiency virus-1 have an increased risk of herpes simplex virus-2 seropositivity and herpes simplex virus reactivation at delivery. Study design: Sixty women infected with human immunodeficiency virus and 8408 other patients who were delivered at the University of Washington between 1989 and 1995 had herpes simplex virus serologic determinations at delivery. Genital herpes simplex virus cultures were obtained for 48 (80%) of the human immunodeficiency virus–infected women and 5567 (66%) of the controls. Logistic regression was used to adjust for possible confounding factors. Results: Forty-five (75%) of human immunodeficiency virus–infected women and 2709 (32%) controls were seropositive for herpes simplex virus-2 ( p < 0.0001). Eight percent of human immunodeficiency virus–infected women and 2% of controls had herpes simplex virus reactivation in labor ( p < 0.05). Conclusions: Infection with herpes simplex virus-2 is common among pregnant women infected with human immunodeficiency virus. Herpes simplex virus reactivation complicates labor in this group more often than in other obstetric patients. The role of herpes simplex virus in perinatal human immunodeficiency virus transmission warrants further study. (Am J Obstet Gynecol 1997;177:450-4.)

Valaciclovir versus aciclovir in patient initiated treatment of recurrent genital herpes: a randomised, double blind clinical trial. International Valaciclovir HSV Study Group.

OBJECTIVE: To compare the efficacy and safety of twice daily valaciclovir with five times daily aciclovir in the treatment of an episode of recurrent genital herpes simplex virus (HSV) infection...