Herpes Simplex Virus Cultures Research Articles

Recurrent genital ulceration: a case of dermatitis herpetiformis.

Case history A 38 year old married Caucasian presented to our department with a two week history of an itchy vesicular penile eruption. At the onset there had been itchiness and erythema followed by oedema and blistering of the prepuce, developing over a 24-hour period. The blisters ruptured leaving superficial ulcers (fig 1). A clinical diagnosis ofherpes was made and swabs from the lesions were sent for herpes simplex virus (HSV) culture. The HSV culture was negative and his initial symptoms improved without any specific treatment, within two weeks. His genital signs and symptoms recurred three times during the next twelve months, before he developed similar lesions over his right shoulder, elbows and sacrum. At this stage the possibility of vesiculo-bullous skin lesions including dermatitis herpetiformis was considered and further investigations were undertaken.

Clinical and Laboratory Study of First Episode Genital Herpes in Singapore

A prospective study of the epidemiology, clinical aspects and laboratory features of first episode genital herpes was carried out in Middle Road Hospital, Singapore between June 1986 and December 1987. A total of 68 patients were studied. There were more women than men in the study population with a female to male ratio of 7.5 to 1. The peak incidence of first episode genital herpes was between 20 and 29 years of age. Specimens from 63 patients yielded positive culture for herpes simplex virus (HSV). HSV-1 was isolated from 21 patients (30.9%), HSV-2 from 41 patients (60.3%) and untyped HSV from one patient (1.5%). True primary genital herpes was diagnosed in 43 patients (68.3%) using a test for complement fixing antibodies to HSV. HSV-1 was found to be the causative agent in 18 (41.9%) of the true primary infections, HSV-2 in 24 (55.8%) and untyped HSV in one (2.3%). In comparison, HSV-1 was isolated in only 2 (10%) of the 20 patients with non-primary first episode genital herpes; HSV-2 was found in 13 patients (65%) and the culture was negative in 5 (25%).

Detection of herpes simplex virus by using A549 cells in centrifugation culture with a rapid membrane enzyme immunoassay

The RAMP herpes simplex virus (HSV) culture confirmation test was compared with immunofluorescence (IF) staining with a specific HSV monoclonal antibody reagent for the detection of HSV in centrifugation culture. The RAMP test detected 47 of 57 IF-positive specimens (sensitivity, 88.6%) and agreed with 217 of 220 IF-negative specimens (specificity, 98.6%). The RAMP test can be performed in less than 15 min and gives an immediate visual result. However, the sensitivity and the false-positive and false-negative results need further investigation.

Identification of herpes simplex viruses by automated profile analysis of viral proteins

In this report we describe a computerized virus identification system based upon the fact that viral proteins, separated by one-dimensional (ID) SDS-PAGE have unique banding patterns (protein profiles). Fifty-eight clinical strains of herpes simplex virus (HSV) were correctly identified by comparing their protein profiles with reference profiles of HSV, other viruses and uninfected cell cultures stored in a computer data base.

Detection of herpes simplex virus by 8 h in shell vial cultures with primary rabbit kidney cells

Shell vial (SV) cultures for herpes simplex virus using primary rabbit kidney cells stained at 8 h after inoculation were compared with 20-h SV cultures as well as conventional tissue culture. Of the 326 clinical specimens examined, conventional culture detected 67, and of these, 61 (91%) and 42 (63%) were detected by 20-h SV cultures and 8-h SV cultures, respectively.

Specific serum IgA, IgG and IgM antibody determination by a modified indirect ELISA-technique in primary and recurrent herpes simplex virus infection

Twenty-three patients with a herpetic infection as diagnosed by a positive culture of herpes simplex virus (HSV) were studied with respect to serological responses of IgA, IgG and IgM antibodies in paired serum samples by an indirect (sandwich) enzyme linked immunosorbent assay (ELISA). Eight of the patients had a primary infection and 15 a recurrent one. In the ELISA test a detergent treated cell lysate of HSV type 1 was used as antigen. In the IgM assay all sera were pretreated with antihuman IgG with the purpose to precipitate IgG of the samples. The conjugate was a F(ab) 2 — fragment of antihuman-IgM. In primary infections all patients had significant titre rises of IgG and presence of high IgM titres in the convalescent serum. IgA antibodies were found in all of them, while titre rises were detected in 5 8 . In recurrent infections titre rises of IgG and IgA antibodies were found in 4 and 5, respectively. Six had detectable IgM in one or both of the paired samples. The IgG titres were higher in recurrent infections than in primary, in contrast to IgM of which much higher titres were found in primary infections. It is concluded that in primary infections a conclusive serological diagnosis was established in all patients, whereas in recurrent infections this was achieved in two of three patients. The indirect ELISA method used for IgM detection was sensitive, reliable and convenient. Interfering rheumatoid factor was effectively eliminated by treatment with antihuman IgG.

Frequency of Asymptomatic Genital Herpes in Pregnant Women at Term

A group of 579 asymptomatic women from Jackson Memorial Hospital, Miami, Florida, and 207 from Kaiser Foundation Hospital in Honolulu, Hawaii, admitted in labor at 37 to 40 weeks of gestation had vaginal cultures for herpes simplex virus (HSV). No cultures were positive and no neonates developed HSV infection. Seven patients gave a history of previous HSV infection in the group from Florida and three from the group in Hawaii. Herpes appears to be a low incidence risk factor in both the populations studied.

Frequency of Asymptomatic Genital Herpes in Pregnant Women at Term

A group of 579 asymptomatic women from Jackson Memorial Hospital, Miami, Florida, and 207 from Kaiser Foundation Hospital in Honolulu, Hawaii, admitted in labor at 37 to 40 weeks of gestation had vaginal cultures for herpes simplex virus (HSV). No cultures were positive and no neonates developed HSV infection. Seven patients gave a history of previous HSV infection in the group from Florida and three from the group in Hawaii. Herpes appears to be a low incidence risk factor in both the populations studied.

Genital Ulceration with Regional Adenopathy

The specific agent causing genital ulceration is difficult to identify without laboratory confirmatory testing including darkfield microscopy, syphilis and lymphogranuloma venereum serology, and culture for Haemophilus ducreyi and herpes simplex virus. Empiric antimicrobial therapy risks masking clinical or serologic diagnosis of syphilis.

Visualization of replicating herpes simplex virus in cervical dorsal root ganglia of mice following explant of individual ganglion onto susceptible indicator cells.

Herpes simplex virus (HSV) establishes a latent state in the sensory ganglia of the peripheral nervous system of its natural or experimental host following primary infection. At various times thereafter, the virus can be reactivated from the latent state whereby it migrates back to the periphery and sometimes initiates a clinical syndrome referred to as recurrent disease. We inoculated mice in the right ear pinna and, following recovery from primary infection, killed the mice at various intervals following either the presence or absence of peripheral stimulations. Explanted cervical dorsal root ganglia yielded HSV in culture and was positive for HSV-like virus particles when viewed with the electron microscope. Hematoxylin and eosin staining showed neuron degeneration, and corresponding HSV-specific immunoperoxidase stains were also positive. The data indicate that ganglionic cells are capable of supporting replicating HSV and that, in vitro, numerous ganglionic cells can be infected simultaneously.

Detection of Herpes Simplex Virus With Primary Rabbit Kidney and an HSV Antigen ELISA

A total of 411 specimens submitted for herpes simplex virus (HSV) was used to evaluate primary rabbit kidney cell cultures and an HSV antigen (culture-amplified) enzyme-linked immunosorbent assay (ELISA). Primary rabbit kidney cell cultures were compared against routinely used cell cultures of primary rhesus monkey kidney (PRMK), primary human embryonic kidney (PHEK), diploid human lung fibroblast (WI-38). and whole human embryo (Flow-5000). The HSV antigen ELISA was evaluated against primary rabbit kidney cultures, which compared favorably with routinely used cell cultures. It detected 91.7% of HSV positives by day 4 and the rest within seven days. Additionally, primary rabbit kidney cell cultures detected two more HSV positives not previously detected by the other cell cultures. The HSV antigen ELISA presented a 90.7% sensitivity and 97.9% specificity in detecting HSV-positive cultures. Combined use of primary rabbit kidney cell cultures and the HSV antigen ELISA has eliminated our use of expensive cell cultures and blind passages for HSV cultures; has decreased reporting time of negative reports from 14 to 4 days, and has been cost-effective in reducing the time and effort needed for HSV identification.

Indirect Immunofluorescent Antibody Diagnosis of Herpes Simplex with Upper Tarsal and Corneal Scrapings

Indirect immunofluorescent techniques were used to diagnose active herpes simplex virus ocular infections in 84 patient observations (41 with ocular lesions suspicious clinically for herpes simplex and 43 with lesions suspicious clinically for other ocular inflammatory conditions). We found indirect immunofluorescent antibody techniques to have a high sensitivity (97%) and specificity (73%) when compared to herpes simplex virus cultures. Similarly, we found the sensitivity (98%) and specificity (77%) of indirect immunofluorescent antibody techniques to be high when compared to the clinical diagnosis of herpes simplex viral infection. Significantly, there were no false negative tests by indirect immunofluorescent techniques. Both corneal and upper tarsal scrapings by indirect immunofluorescence were used and the upper tarsal scrapings were an excellent source of cells exhibiting herpes simplex virus antigens. All cases in which corneal scrapings were positive by indirect immunofluorescence for herpes simplex were also positive by upper tarsal scrapings, although the converse was not true.

A rabbit model of focal herpes simplex encephalitis.

Results from studies designed to create a model of focal encephalitis caused by herpes simplex virus (HSV) are reported. Anesthetized rabbits underwent exposure and inoculation of the olfactory bulb with three different doses of a wild-type HSV. Lethal infection resulted in 69% of the animals, without evidence for a dose-response relationship. Necropsy specimens obtained on or before day 10 after inoculation routinely yielded HSV in culture. In 76% of the animals with positive cultures for virus, these cultures originated exclusively or primarily from the pyriform (or temporal) cortex and frontal lobes. Virus could not be cultured from animals killed more than two weeks after inoculation. Histological examination of brains obtained three or more days after inoculation demonstrated evidence of viral infection, with more severe involvement of temporal cortex than of the surrounding brain in 80%. Immunohistochemical demonstration of viral antigens persisted for up to three weeks after inoculation.

Herpes simplex virus testing of an obstetric population with an antigen enzyme-linked immunosorbent assay

Commercial herpes simplex virus antigen enzyme-linked immunosorbent assay kits were compared to conventional culture for herpes simplex virus with 3237 genital specimens from obstetric patients. These rapid enzyme-linked immunosorbent assay tests had a sensitivity of 34.3% and specificity of 98.1% with primarily cervical specimens from asymptomatic patients. Specimens from vulvar swabs had higher positive rates than those from cervical swabs from the same patient, whether symptomatic or asymptomatic with both culture and enzyme-linked immunosorbent assay. Fifty-six women had enzyme-linked immunosorbent assay false positive tests; use of enzyme-linked immunosorbent assay testing alone could have resulted in 1.7% unnecessary cesarean deliveries. Six cases of neonatal herpes simplex virus infection were identified; two of the mothers had negative cervical cultures the week of delivery preceded by positive vulvar cultures.

Association of concurrent herpes simplex virus and cytomegalovirus with detrimental effects after renal transplantation.

Herpes simplex virus (HSV) and cytomegalovirus (CMV) infections occur frequently after renal transplantation. To determine the effect of these infections on long-term prognosis, we reviewed the charts of 558 patients who underwent primary renal transplantation between Jan 1, 1974 and Dec 31, 1978, at the University of Minnesota Hospitals, Minneapolis, for cultural evidence of these infections. The presence of HSV alone appeared to have a minimal effect on either patient or allograft survival. Positive CMV cultures were associated with decreased patient and allograft survival. Patients with HSV and CMV infections had further reduced patient and allograft survival. Lethal CMV syndrome and associated severe bacterial and fungal infections were common and frequently were the immediate cause of death in patients who had concomitant positive HSV and CMV cultures. Concurrent HSV and CMV infections in renal transplant patients may be a grave prognostic indicator, being associated with enhanced mortality and renal allograft loss.

Rapid detection and identification of herpes simplex virus in cell culture by a direct immunoperoxidase staining procedure.

Cell monolayers were inoculated with 169 fresh and 76 previously frozen clinical specimens and examined for the presence of herpes simplex virus (HSV) by noting the appearance of characteristic cytopathic effect (CPE) and by using direct immunoperoxidase (IP) stain for viral antigen. HSV was detected by IP staining in 40 of 169 (23.7%) monolayers and by CPE in 39 of 169 (23.1%) monolayers inoculated with fresh specimens. All 40 isolates were detected and confirmed by IP staining within 24 h. Although 39 of 40 isolates were detected by CPE, only 9 of 39 (23%) were positive within 24 h. CPE was observed at 2.7 days on the average, but 4 days were required before 90% of the cultures were positive and more than 5 days were required before all HSV isolates were recognized. Similar results were observed for frozen specimens. HSV was detected earlier with IP staining, which demonstrated more extensive infection of cell monolayers inoculated with titrated fresh culture isolates and clinical specimens than did CPE. IP staining reduces the amount of time required for detection and identification of HSV in culture, is readily adaptable for use in the clinical laboratory, and permanent stained preparations can be made.

Characteristics and management of pregnancy in women with genital herpes simplex virus infection

The natural history of genital herpes simplex virus (HSV) infections was investigated in 83 pregnancies in 78 women, and the information was used to determine the need for cesarean section in these women. We studied 163 recurrent episodes with HSV cultures from the cervix and from vulvar lesions every 1 to 3 days. Cervical HSV cultures were obtained weekly from asymptomatic women beginning at 32 weeks' gestation, and 14/462 (3.03%) of these cultures were positive. Cervical cultures obtained during culture-positive vulvar recurrences demonstrated concomitant cervical HSV shedding in 25/165 (15.2%) cultures. Mean duration of 26 genital HSV recurrences was 4.6 ± 2.8 days with a range of 1 to 13 days. The mean interval between culture-positive HSV recurrences was 59.2 ± 42.1 days, but many (14/76 = 18%) intervals were less than 21 days. Viral cultures were already positive in 92.3% of cases after 4 days' incubation, so they could be used effectively to determine route of delivery. Following a cautious set of criteria for vaginal delivery in these women, 69.1% were delivered of their infants vaginally and no neonatal morbidity caused by HSV was encountered. Information about the natural history of genital HSV infections obtained from frequent third-trimester viral cultures can be used to manage pregnancy and will reduce the need for cesarean section while avoiding neonatal HSV morbidity.

The incidence and outcome of asymptomatic herpes simplex genitalis in an obstetric population

In a prospective study of an obstetric population, 28 of 488 (4%) routine asymptomatic patients had positive cultures for herpes simplex virus from their genital tract. Cesarean section was carried out if the virus was present in the genital tract at the time of delivery. In one instance, a culture which had reverted to negative became positive again. No cases of neonatal herpes were reported in the time of the study. As yet, the infectivity of these subclinical cases is unclear, and the incidence of neonatal herpes is not as high as would be expected from the study. Further studies need to be carried out to evaluate the exact risk of the asymptomatic herpesvirus shedder to the infant at birth.

Herpes Simplex Antigen in Immune Complexes of Patients With Erythema Multiforme

Serum samples from patients with erythema multiforme (EM) were examined for the presence of herpes simplex virus (HSV) antigen in immune complexes using the Raji cell radioimmunoassay. Raji-cell-bound immune complexes from eight of 12 patients with EM after HSV infection and four of four patients with EM of uncertain cause had increased anti-HSV binding, while patients with EM after drug exposure and patients with recurrent HSV infections without EM had binding in the same range as controls. Viral cultures for HSV of immune complexes eluted from Raji cells were negative. Sucrose density gradient ultracentrifugation studies of serum samples of patients with EM after HSV infection showed HSV antigen in large molecular weight fractions. The HLA typing of lymphocytes from 16 patients with EM after HSV infection was not different from that of controls. Immune complexes composed of antibody and HSV antigen are present in serum samples of patients with EM after HSV infection and some cases of EM of uncertain cause and may mediate the pathogenesis of these disorders.

Recurrent genital herpes simplex virus infection in pregnancy: Infant outcome and frequency of asymptomatic recurrences

Eighty pregnant patients with a history of recurrent genital herpes simplex virus (HSV) infection were followed up with frequent genital examinations and cultures for HSV during their pregnancies. Recurrences of genital HSV during pregnancy were documented in 67 patients. Ninety-six percent of these had external genital lesions noted at some time during their pregnancy. One hundred eighty-six (93%) of the 199 recurrences of the disease were associated with external genital lesions. Of 13 episodes of asymptomatic viral shedding documented in this patient population, six were from the vulva alone, five were from the cervix alone, and two were from both the vulva and the cervix. Of 11 episodes of cervical HSV shedding, seven were asymptomatic and four were associated with external symptomatic lesions. Despite frequent recurrences of genital HSV infection during pregnancy, all study patients were delivered at term, and although they had a high rate of delivery by cesarean section (32.5%), the outcome of their infants was good.