Persistence of varicella-zoster virus viraemia in patients with herpes zoster

BackgroundHerpes zoster is caused by the reactivation of varicella-zoster virus from sensory neurons. The commonest complication following zoster is chronic pain termed post herpetic neuralgia.ObjectivesTo investigate the dynamics of VZV viraemia and viral load following the resolution of zoster and its relationship to PHN development.Study designBlood samples were collected at baseline, 1 month, 3 months and 6 month from a prospective study of 63 patients with active zoster. Quantification of VZV DNA in whole blood was performed using a real-time PCR assay.ResultsDuring acute zoster, all patients had detectable VZV DNA in their blood. VZV DNA remained detectable in the blood of 91% of patients at 6 months although levels declined significantly (p < 0.0001). A history of prodromal symptoms (p = 0.005) and severity of pain at baseline (p = 0.038) as well as taking antivirals (p = 0.046) and being immunocompromised (p = 0.043) were associated, with longer time to recovery from PHN. Viral DNA loads were consistently higher in patients with risk factors for PHN and higher viral DNA loads over time were associated with longer time to recovery (p = 0.058 overall and 0.038 in immunocompetent).ConclusionsBased on these observations we hypothesise that VZV replication persists following acute shingles and that higher viral DNA loads contribute to the risk factors for PHN.     

Detection of varicella-zoster virus DNA in throat swabs of patients with herpes zoster and on air purifier filters

Zoster patients are considered to be less contagious than those with varicella because their infectious lesions are localized. However, it is not known when the spread of varicella-zoster virus (VZV) from zoster patients begins, how long it continues, and how far the virus spreads from the zoster patients. Twelve cases of hospitalized zoster patients were studied. The polymerase chain reaction (PCR) was used to detect VZV DNA in samples taken from the surface of their eruptions, throats, and the air purifier filters in their rooms. In all patients, VZV DNA was detected in the samples from eruptions. VZV DNA was detected in 8 of 12 patients from the throats. VZV DNA was detected for 9 of 12 patients from the filter samples. This study shows the possibility of a wide distribution of VZV DNA to the environment from infected patients. VZV may be excreted from cutaneous eruptions or from the throats of patients.     

Rapid detection of herpes simplex virus and varicella-zoster virus infections by real-time PCR

The herpes simplex viruses types 1 and 2 (HSV-1 and HSV-2) and varicella-zoster virus (VZV) can cause life-threatening infections of the central nervous system and lead to severe infections in immunocompromised subjects and newborns. In these cases, rapid diagnosis is crucial. We developed three different real-time PCR assays based on TaqMan chemistry for the LightCycler instrument to detect HSV-1, HSV-2, and VZV. When the TaqMan assays were compared to our in-house nested PCR assays, the test systems had equal sensitivities of 相似文献   

Detection of varicella-zoster virus DNA by polymerase chain reaction in the cerebrospinal fluid of patients suffering from neurological complications associated with chicken pox or herpes zoster.

The polymerase chain reaction (PCR) was used to detect varicella-zoster virus (VZV) DNA in the cerebrospinal fluid of patients with VZV infection associated with neurological symptoms. Positive results were obtained in three of five children with post-chicken pox cerebellitis and in seven of seven herpes zoster patients with neurological symptoms. The PCR thus provides a useful tool for the early diagnosis of VZV-associated neurological disease.     

Persistence of varicella-zoster virus DNA in blood mononuclear cells of patients with varicella or zoster

Varicella-zoster virus (VZV) DNA was detectable by in-situ hybridization in blood mononuclear cells (MNCs) of patients with varicella or zoster for 2–56 days after the onset of a rash. VZV DNA was present in many MNCs from one acute varicella patient 2 days after the onset of the rash and was rarely found in MNCs during acute zoster, convalescent zoster, and convalescent varicella. The morphology of MNCs containing VZV was heterogenous, although most viral-DNA-containing MNCs were large monocytoid cells. Serial examination of blood MNCs from one adult with varicella revealed VZV DNA up until 8 weeks, but not 16 weeks, after the appearance of the rash; parallel studies in four zoster patients showed VZV DNA up until 3 weeks, but not later than 7 weeks after the appearance of the rash. These results indicate that MNCs become infected with VZV during the primary encounter with VZV (varicella) and during reactivation (zoster) and that infection continues for weeks after the onset of the skin rash. Furthermore, the detection of VZV DNA in blood MNCs of uncomplicated zoster patients coincides with the period during which these patients experience pain.     

Quantitation of varicella-zoster virus DNA in patients with Ramsay Hunt syndrome and zoster sine herpete

Varicella-zoster virus (VZV) reactivation causes facial nerve palsy in Ramsay Hunt syndrome (RHS) and zoster sine herpete (ZSH) with and without zoster rash, respectively. In the present study, we analyzed the VZV DNA copy number in saliva samples from 25 patients with RHS and 31 patients with ZSH using a TaqMan PCR assay to determine differences in the viral load between the two diseases. VZV copy number in saliva peaked near the day of the appearance of zoster in patients with RHS. Consequently, VZV DNA was less frequently detected in patients with RHS who exhibited facial palsy several days after the appearance of zoster. These findings suggest that the VZV load in saliva samples reflects the kinetics of viral reactivation in patients with RHS. In addition, VZV DNA was equally detected in saliva from patients with RHS and ZSH, and there was no significant difference in the highest viral copy number between patients with RHS and those with ZSH. The VZV load does not appear to reflect a major difference between RHS and ZSH.     

Detection of varicella-zoster virus DNA in peripheral mononuclear cells from patients with Ramsay Hunt syndrome or zoster sine herpete

On the basis of alterations in varicella-zoster virus (VZV) antibody titers, it appears that Bell's palsy in some patients could be associated with VZV reactivation, that is, zoster sine herpete. To obtain stronger evidence of this association, polymerase chain reaction (PCR) was used to detect VZV DNA in auricular lesions or peripheral blood mononuclear cells (PBMCs) from Bell's palsy or Ramsay Hunt syndrome patients. VZV DNA was detected in the auricular lesions of Ramsay Hunt syndrome, in PBMCs from 2 Ramsay Hunt syndrome patients, and in 4 of 17 samples from 16 Bell's palsy patients. Three of these four positive patients were thought to have zoster sine herpete because of hearing difficulty, vertigo, and pain. VZV IgM antibodies were positive in 1 of the 2 patients with Ramsay Hunt syndrome, and in 2 of the 17 samples from the Bell's palsy patients. VZV IgG antibody titers during the acute phase were significantly higher in the patients positive for the PCR or VZV IgM antibody than in those negative for them. These findings provide evidence that Bell's palsy in some patients could be associated with VZV reactivation. J. Med. Virol. 56:359–363, 1998 . © 1998 Wiley-Liss, Inc.     

Reactivity of varicella-zoster virus subunit antigens in enzyme-linked immunosorbent assay to sera from varicella,zoster, and herpes simplex virus infections

Serological responses to varicella-zoster virus (VZV) subunit antigens, such as capsid, envelope, and soluble (S) antigens, in patients with VZV and herpes simplex virus (HSV) infections were studied by comparing with responses to virion (V) antigens using an enzyme-linked immunosorbent assay (ELISA). S antigen, prepared by concentrating supernatant of VZV or HSV type 1 (HSV-1)-infected cell culture fluid, reacted strongly to sera from patients with secondary infection but reacted poorly to those from patients with a primary infection of VZV or HSV. Antibody titers to VZV-S antigen persisted for a long period in patients with VZV infections. Patients infected with VZV showed antibody increase to HSV-1, when tested by complement fixation or complement-enhanced neutralization test, in cases with a history of prior HSV infection. However, such a cross-reaction was only observed to a minor extent in ELISA test using S antigen. S antigen reaction was stronger in secondary infections in tests with various subunit antigens. Almost no cross-reactivity was observed in an immunoblotting test with S antigen. Differentiation between infections with either varicella or zoster or HSV can be made by comparison of antibody responses to V and S antigens.     

Specificity of skin test with varicella-zoster virus antigen in varicella-zoster and herpes simplex virus infections.

Specificity of the skin test with varicella-zoster virus (VZV) antigen was examined in guinea pigs infected with herpes simplex virus (HSV) type 1 or VZV and in children with a history of HSV infection who developed varicella. Infected guinea pigs responded positively only to homologous virus. No cross-reaction between HSV and VZV was detected in the skin test, as well as in the neutralization test in infected guinea pigs, suggesting that the VZV skin test is specific for immunity to VZV infection. Twelve children were infected with HSV during an HSV epidemic and subsequently developed varicella in institutional settings. During the 2.5-month period between the HSV and VZV infections, the immune status of the children to VZV was negative both in the skin test and in the antibody test, although antibody to HSV was detected by an immune adherence hemagglutination test. After VZV infection, all responded positively both in the skin test and in the antibody test (immune adherence hemagglutination test) to VZV. These results suggest that the VZV skin test is specific for immunity to VZV infection, not cross-reactive to HSV infection in humans. This specificity will be of value in screening susceptibility or immunity to VZV, irrespective of prior HSV infection.     

Comparison of the DNAs of varicella-zoster viruses isolated from clinical cases of varicella and herpes zoster

The DNA of varicella-zoster virus (VZV) has been characterized by sucrose gradient sedimentation and by isopycnic banding in CsCl. Comparisons of the DNAs from different clinical isolates have been made by mixing radiolabeled DNAs prior to centrifugation. VZV-DNA sedimented just behind T4 DNA on neutral sucrose gradients. Thus VZV-DNA has a molecular weight of about 100 x 10⁶. There were no distinguishable differences in the sedimentation behavior in neutral sucrose gradients of the DNAs from several clinical VZV isolates. The buoyant density in CsCl of the DNA of VZV isolated from varicella was reproducibly slightly lighter than the buoyant density of the DNA of VZV isolated from herpes zoster.     

Different genotype pattern of varicella-zoster virus obtained from patients with varicella and zoster in Germany

The general use of the varicella vaccine requires the surveillance of varicella-zoster virus (VZV) strains in patients infected with VZV. This paper reports the data achieved from a prospective study of genotyping VZV in Germany, analyzing the restriction fragment length polymorphism (RFLP) of the open reading frames (ORF) 38, 54, and 62 as well as the polymorphism of the R5 repeat region. The study included 177 patients with varicella. Seventy-eight patients with zoster served as controls. Results revealed that 78% of VZV strains in patients with varicella had the genetic profile of the dominant wild-genotype occurring in Europe and 22% had the markers of African or Asian strains. Varicella patients with the profile of African or Asian strains were significantly younger than patients with varicella caused by the dominant genotype. By contrast, all zoster patients exhibited strains representing the majority of wild-type strains in Europe. In conclusion, VZV strains from patients with varicella have a significantly higher genetic variability than viral strains from zoster patients. Since variants with the markers of African or Asian strains could only be found in young children with chickenpox, the results suggest a changing scene of VZV genotypes in Germany. As reasons, the spread of viruses, which may be imported originally by persons immigrating from warmer climates, or the recombination between wild-and vaccine-type viruses have to be considered.     

Rapid DNA diagnosis of herpes simplex virus serotypes.

The presence of nucleotide sequences specific for each of herpes simplex virus (HSV) serotypes was demonstrated. These sequences were applied for dot DNA-DNA hybridization and for PCR for rapid DNA diagnosis of HSV infections. These sequences were found by molecular cloning of HSV-DNA fragments after digestion of DNA by KpnI enzyme. The type 1-specific sequence was found around the 5' end of BamHI B-fragment in the L region of type 1 DNA (corresponds to alpha gene 27, promoter-regulatory region) and the type 2-specific sequence was around the junction region of the L and S of type 2 DNA (corresponds to a' sequence). Both simple dot blot hybridization and PCR of HSV DNA's, employing these type-specific nucleotide sequences, were proven to be much more useful than immunofluorescence in terms of type-specific diagnosis of HSV infections.     

Simian varicella virus DNA shares homology with human varicella-zoster virus DNA

Delta herpesvirus (DHV) and Medical Lake Macaque (MLM) virus are cell-associated simian herpesviruses that cause varicella-like disease in nonhuman primates, and are antigenically related to human varicella-zoster virus (VZV). The results of studies designed to determine if homology exists between the DNA of DHV and MLM and the DNA of VZV are reported here. Southern blot hybridizations conducted at Tm-20 degrees did not detect DNA homology between the VZV and simian varicella virus genomes. However, under conditions of lower stringency (Tm-36 degrees and Tm-43 degrees), VZV DNA probes hybridized to specific HindIII fragments within DNA isolated from simian varicella virus-infected cells. Under similar hybridization conditions, DNA homology was not detected between VZV DNA and herpes simplex virus DNA. Further studies using cloned VZV DNA HindIII fragments as probes suggested that the homology between VZV DNA and DHV DNA is distributed across the viral genomes. These results demonstrate that the genomes of VZV and simian varicella virus share regions of conserved nucleotide sequences, and indicate a close evolutionary relationship between VZV and simian varicella viruses. In addition, the studies show that the DHV and MLM strains of simian varicella virus are more closely related to each other than to human VZV.     

Sensitive and rapid detection of herpes simplex virus and varicella-zoster virus DNA by loop-mediated isothermal amplification

Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method in which reagents react rapidly and efficiently, with a high specificity, under isothermal conditions. We used a LAMP assay for the detection of herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), and varicella-zoster virus (VZV). The virus specificities of primers were confirmed by using 50 HSV-1, 50 HSV-2, and 8 VZV strains. The assay was performed for 45 min at 65 degrees C. The LAMP assay had a 10-fold higher sensitivity than a PCR assay. An analysis of nucleotide sequence variations in the target and primer regions used for the LAMP assay indicated that 3 of 50 HSV-1 strains had single nucleotide polymorphisms. No HSV-2 or VZV strains had nucleotide polymorphisms. Regardless of the sequence variation, there were no differences in sensitivity with the HSV-1-specific LAMP assay. To evaluate the application of the LAMP assay for clinical diagnosis, we tested clinical samples from 40 genital herpes patients and 20 ocular herpes patients. With the LAMP assay, 41 samples with DNA extraction and 26 direct samples without DNA extraction were identified as positive for HSV-1 or HSV-2, although 37 samples with DNA extraction and just one without DNA extraction were positive by a PCR assay. Thus, the LAMP assay was less influenced than the PCR assay by the presence of inhibitory substances in clinical samples. These observations indicate that the LAMP assay is very useful for the diagnosis of HSV-1, HSV-2, and VZV infections.     

Disseminated herpes simplex virus and varicella zoster virus coinfection in a patient taking thalidomide for relapsed multiple myeloma

Disseminated herpes simplex virus (HSV) and varicella zoster virus (VZV) have been reported individually in immunosuppressed adults. We present a case of coinfection with disseminated HSV and VZV infection in a patient taking thalidomide for relapsed multiple myeloma. This is the first report of opportunistic infection associated with thalidomide.     

Polymorphism of herpes simplex virus type 1 and 2 DNA from laboratory strains and clinical material from patient with genital herpes

Polymerase chain reaction (PCR), that can amplify a fragment of the DNA-polymerase gene of 4 herpes viruses, i.e. herpes simplex viruses, type 1 (HSV-1), herpes simplex viruses, type 2 (HSV-2), Epstein-Barr virus and cytomegalovirus, was made use of to study the genetic polymorphism of HSV-1 and HSV-2 strains. The obtained amplicons were analyzed by the method of restriction-size fragments' polymorphism (RSFP) with restrictases Rsal, Taql and Hinfl. Four HSV-1 strains had an identical restriction profile. Strain G (HSV-2) also displayed the expected restriction profiles, however, contradictory results were obtained for strain BH (HSV-2): the restriction profiles with restrictases Hinfl and Rsal corresponded to HSV-2, and the restriction profile with Taql corresponded to HSV-1. The sequencing of appropriate fragments of strains G and BH revealed a dot-type mutation localized in Taql restriction site. The thus worked out PCR was used jointly with RSFP in the genotyping of 75 urogenital samples obtained from women with genital herpes who were treated at Moscow patient-care facilities. HSV-1 and HSV-2 were detected in 18 (24%) and 57 (76%) of samples, respectively. No changes were registered in the restriction profile for HSV-2 among the investigated samples and all of them had the restriction profile similar to that of strain G. The conclusion is that genital herpes associated with HSV-2 is genetically stable within its Moscow population.     

Serum immunoglobulin A antibody to varicella-zoster virus in subjects with primary varicella and herpes zoster infections and in immune subjects.

Immunoglobulin A (IgA) antibodies to varicella-zoster virus (VZV) were measured in sera from subjects with acute varicella and herpes zoster, VZV-immune subjects remote from infection, and recipients of a live attenuated varicella vaccine, using a solid-phase radioimmunoassay. Primary infection with VZV was associated with early production of IgA antibodies. Among 36 subjects with varicella tested 1 to 5 days after onset, 22 had detectable IgA, and all of the negative sera were obtained before day 3 of the varicella exanthem. VZV IgA was detected in one of three sera obtained more than 60 days after onset of the illness. Four of five sera obtained from subjects within 1 week of the onset of herpes zoster had measurable levels of IgA. Between 1 and 4 weeks after onset of zoster, all 10 subjects tested had detectable IgA to VZV. VZV IgA was detected as late as 63 days after the onset of herpes zoster. Of 10 vaccine recipients, 5 developed VZV IgA which was detected as early as 4 weeks and persisted for as long as 16 weeks after vaccination. VZV IgA was not detected in sera from 42 children who had no detectable IgG antibody to VZV. VZV IgA was found on only 3 of 23 sera from adults who had varicella more than 20 years before.     

Detection of varicella-zoster virus and herpes simplex virus by the polymerase chain reaction with degenerate primers

Varicella-zoster virus (VZV) and herpes simplex virus (HSV) are human pathogens of significance involved in multiple diseases with either typical or atypical clinical features. In neonates and immunocompromised patients these alphaherpesviruses may cause life-threatening diseases such as encephalitis. Detection of VZV by virus culture is difficult. Polymerase chain reaction (PCR) is quicker and more sensitive and applicable in most clinical microbiological laboratories. Using degenerate primers, glycoprotein B (gB) DNA was amplified from all alphaherpesvirus field strains present in clinical samples. The amplification of gB allowed virus typing of VZV, HSV-1 and HSV-2 using restriction enzyme digestion of the PCR products. Degenerate primers can replace conventional primers in diagnostic PCR without loss of sensitivity and specificity.