Herpesvirus DNA Research Articles

Prevalence of Herpes Simplex and Varicella-Zoster Virus DNA in Corneal Grafts Is Higher than Expected.

(1) To determine the prevalence of herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2), varicella-zoster virus (VZV), and cytomegalovirus (CMV) DNA in donor corneas; (2) To evaluate the clinical outcome of the grafts with viral DNA and to compare donors with and without viral DNA. We analyzed data from all donors and recipients who underwent penetrating keratoplasty (PK) or Descemet membrane endothelial keratoplasty (DMEK) between September 2022 and March 2023. Donor corneoscleral rims and excised recipients' corneal buttons were tested for the presence of HSV-1, HSV-2, VZV, and CMV DNA by polymerase chain reaction (PCR). The results were known 2-3 days after the surgery. We closely followed up on patients whose grafts tested positive for viral DNA. We compared the medical histories of donors with and without viral DNA. We included 85 corneas from 67 donors. Seven (8.2%) donor corneas tested positive for HSV-1 (n = 3) or VZV (n = 4) DNA. We did not detect any HSV-2 or CMV DNA. In the postoperative follow-up of patients with positive PCR, a graft failure was observed in one and infections in two eyes. Re-operation was necessary in three of these cases (42.9%). Patients without herpes DNA in the donor cornea needed reoperation in 7.7% of the cases. Cultural duration, the cause of the donor's death, and the death-to-explantation interval did not differ significantly between donors with and without viral DNA. Additionally, 3 of the 7 (42.9%) donors with positive PCR were in a septic status at the time of death, compared to 21 of the 78 (26.9%) donors with negative PCR (p = 0.52). The prevalence of herpes DNA in the donor corneas was 8.2% and thus higher than previously reported. We did not notice any evidence for a donor-to-host transmission, but a higher rate of postoperative complications in recipients of the grafts with viral DNA. The donors with and without herpetic DNA did not differ significantly regarding systemic diagnoses or cultural conditions, but sepsis was more frequent in the group with viral DNA.

Higher frequency of Human herpesvirus-6 (HHV-6) viral DNA simultaneously with low frequency of Epstein-Barr virus (EBV) viral DNA in a cohort of multiple sclerosis patients from Rio de Janeiro, Brazil.

Multiple sclerosis (MS) is a chronic neuroinflammatory and neurodegenerative disease of the central nervous system (CNS). The etiology of MS is not well understood, but it's likely one of the genetic and environmental factors. Approximately 85% of patients have relapsing-remitting MS (RRMS), while 10-15% have primary progressive MS (PPMS). Epstein-Barr virus (EBV) and Human herpesvirus 6 (HHV-6), members of the human Herpesviridae family, are strong candidates for representing the macroenvironmental factors associated with MS) pathogenesis. Antigenic mimicry of EBV involving B-cells has been implicate in MS risk factors and concomitance of EBV and HHV-6 latent infection has been associated to inflammatory MS cascade. To verify the possible role of EBV and HHV-6 as triggering or aggravating factors in RRMS and PPMS, we compare their frequency in blood samples collected from 166 MS patients. The presence of herpes DNA was searched by real-time PCR (qPCR). The frequency of EBV and HHV-6 in MS patients were 1.8% (3/166) and 8.9% (14/166), respectively. Among the positive patients, 100% (3/3) EBV and 85.8% (12/14) HHV-6 are RRMS and 14.4% (2/14) HHV-6 are PPMS. Detection of EBV was 1.2% (2/166) and HHV-6 was 0.6% (1/166) in blood donors. About clinical phenotype of these patients, incomplete multifocal myelitis, and optic neuritis were the main CNS manifestations. These are the first data about concomitant infection of these viruses in MS patients from Brazil. Up to date, our findings confirm a higher prevalence in female with MS and a high frequency of EBV and HHV-6 in RRMS patients.

Presence of Multiple Herpesvirus Variants in Australian Flying Foxes (Pteropus spp.).

Herpesviruses have been detected in bat species from several countries, with a limited number of studies examining herpesviruses in Pteropus spp. (flying foxes) and no investigation of herpesviruses in Australian flying foxes. We examined the presence and prevalence of herpesviruses in the four mainland Australian flying fox species. A nested PCR targeting highly conserved amino acid motifs in the DNA polymerase (DPOL) gene of herpesviruses was used to analyze 564 samples collected from 514 individual Pteropus scapulatus, Pteropus poliocephalus, Pteropus alecto, and Pteropus conspicillatus. The prevalence of herpesvirus DNA in blood, urine, oral, and fecal swabs from the four species was 17% in P. scapulatus, 11% in P. poliocephalus, 10% in P. alecto, and 9% in P. conspicillatus (31% in P. conspicillatus spleen tissue). Five putative novel herpesviruses were detected. Following PCR amplicon sequence analysis, four of the herpesviruses grouped phylogenetically with the gammaherpesviruses, with nucleotide identities between 79% and 90% to gammaherpesviruses from Asian megabats. A betaherpesvirus was detected in P. scapulatus with 99% nucleotide identity to the partial DPOL gene sequence of an Indonesian fruit bat betaherpesvirus. This study lays the foundation for future epidemiology research of herpesviruses in Australian Pteropus spp. and adds to the discussion of hypotheses surrounding the evolutionary epidemiology of bat-borne viruses on a global scale.

Surveillance, Isolation, and Genetic Characterization of Bat Herpesviruses in Zambia.

Bats are of significant interest as reservoirs for various zoonotic viruses with high diversity. During the past two decades, many herpesviruses have been identified in various bats worldwide by genetic approaches, whereas there have been few reports on the isolation of infectious herpesviruses. Herein, we report the prevalence of herpesvirus infection of bats captured in Zambia and genetic characterization of novel gammaherpesviruses isolated from striped leaf-nosed bats (Macronycteris vittatus). By our PCR screening, herpesvirus DNA polymerase (DPOL) genes were detected in 29.2% (7/24) of Egyptian fruit bats (Rousettus aegyptiacus), 78.1% (82/105) of Macronycteris vittatus, and one Sundevall's roundleaf bat (Hipposideros caffer) in Zambia. Phylogenetic analyses of the detected partial DPOL genes revealed that the Zambian bat herpesviruses were divided into seven betaherpesvirus groups and five gammaherpesvirus groups. Two infectious strains of a novel gammaherpesvirus, tentatively named Macronycteris gammaherpesvirus 1 (MaGHV1), were successfully isolated from Macronycteris vittatus bats, and their complete genomes were sequenced. The genome of MaGHV1 encoded 79 open reading frames, and phylogenic analyses of the DNA polymerase and glycoprotein B demonstrated that MaGHV1 formed an independent lineage sharing a common origin with other bat-derived gammaherpesviruses. Our findings provide new information regarding the genetic diversity of herpesviruses maintained in African bats.

Porcine Mx proteins inhibit pseudorabies virus replication through interfering with early gene synthesis

Pseudorabies virus (PRV) is an enveloped, linear double-stranded DNA herpesvirus that resulted in huge financial losses to the swine industry. In addition to vaccination, the development of antiviral molecules is also a beneficial supplement to the control of Pseudorabies (PR). Although our previous studies have shown that porcine Mx protein (poMx1/2) significantly inhibited the proliferation of RNA virus, it was unknown whether poMx1/2 could inhibit porcine DNA virus, such as PRV. In this study, it was investigated the inhibitory effect of porcine Mx1/2 protein on PRV multiplication. The results showed that both poMx1 and poMx2 had anti-PRV activities, which required GTPase ability and stable oligomerization. Interestingly, the two GTPase deficient mutants (G52Q and T148A) of poMx2 also had the antiviral ability against PRV, which was consistent with previous reports, indicating that these mutants recognized and blocked the viral targets. Mechanistically, the antiviral restriction of poMx1/2 came from their inhibition of the early gene synthesis of PRV. Our results for the first time shed light on the antiviral activities of two poMx proteins against DNA virus. The data from this study provide further insights to develop new strategies for preventing and controlling the diseases caused by PRV.

Detection of Equine Papillomaviruses and Gamma-Herpesviruses in Equine Squamous Cell Carcinoma.

Squamous cell carcinoma (SCC) seriously compromises the health and welfare of affected horses. Although robust evidence points to equine papillomavirus type 2 (EcPV2) causing genital lesions, the etiopathogenesis of equine SCC is still poorly understood. We screened a series of SCCs from the head-and-neck (HN), (peri-)ocular and genital region, and site-matched controls for the presence of EcPV2-5 and herpesvirus DNA using type-specific EcPV PCR, and consensus nested herpesvirus PCR followed by sequencing. EcPV2 DNA was detected in 45.5% of HN lesions, 8.3% of (peri-)ocular SCCs, and 100% of genital tumors, whilst control samples from tumor-free horses except one tested EcPV-negative. Two HNSCCs harbored EcPV5, and an ocular lesion EcPV4 DNA. Herpesvirus DNA was detected in 63.6%, 66.6%, 47.2%, and 14.2% of horses with HN, ocular, penile, and vulvar SCCs, respectively, and mainly identified as equine herpesvirus 2 (EHV2), 5 (EHV5) or asinine herpesvirus 5 (AsHV5) DNA. In the tumor-free control group, 9.6% of oral secretions, 46.6% of ocular swabs, 47% of penile samples, and 14.2% of vaginal swabs scored positive for these herpesvirus types. This work further highlights the role of EcPV2 as an oncovirus and is the first to provide information on the prevalence of (gamma-)herpesviruses in equine SCCs.

Герпесвирусные инфекции в формировании эпизодов ОРВИ у детей

Aim: to evaluate the detection rate of the herpesvirus activity in children with acute respiratory viral infection (ARVI) signs for determination of the optimal treatment tactics. Patients and Methods: the study included 1120 children aged 3 to 18 years (mean age 7.48±3.91 years) who were observed on an outpatient basis (n=650) or hospitalized (n=379) due to the lack of improvement on the 3rd day after the first ARVI symptoms onset (n=1029) or due to the detected complication (n=91). The herpesvirus DNA was detected in throat swabs by PCR. Enzyme immunoassay was used to evaluate the levels of IgG, IgM, and IgA antibodies. Results: the presence of IgG antibodies to at least one of the herpes viruses was detected in 1097 (97.9%) of patients: in the children group over 6 years old in 723 (100%) patients. The presence of herpesvirus DNA in smears from the posterior pharyngeal wall and tonsils was detected in 234 (20.9%) children: 178 (15.9%) — isolated, 56 (5.0%) — in various combinations. IgM antibodies to Epstein-Barr virus and cytomegalovirus were detected in all cases in the herpesvirus DNA presence in the blood. The herpes simplex virus DNA has not been detected in any case. However, antibodies were detected in 63 (5.6%) children: IgA in 57 (5.1%) and IgM in 6 (0.5%) children. Conclusions: in children with ARVI symptoms, signs of an active infection caused by one or another type of herpes were detected in 26.5% of cases, and in 5% — their combination. The high prevalence of the herpesvirus activity comparable with the detection rate of seasonal respiratory viral infections, along with the adverse effect of herpesvirus infection on the immune system with the severe somatic pathology probability, necessitates the use of a drug with a wide spectrum of antiviral action for the ARVI signs in children — inosine pranobex. In the presence of complications caused by herpesvirus, inosine pranobex does not cause negative trend of antibody activity indicators, inosine pranobex, Groprinosin® KEYWORDS: HHV-6, human herpesvirus type 6, herpes simplex virus, Epstein-Barr virus, herpes, herpesvirus infection, herpes infection, ARVI, cytomegalovirus, inosine pranobex, Groprinosin. FOR CITATION: Ruzhentsova T.A., Popova R.V., Garbuzov A.A., Shushakova E.K., Khalyastov I.N., Meshkova N.A. Herpesvirus infections in ARVI episodes in children. Russian Journal of Woman and Child Health. 2023;6(4):399–404 (in Russ.). DOI: 10.32364/2618-8430- 2023-6-4-12.

Эпштейна — Барра вирус — ведущая причина гипертрофии глоточной миндалины у детей

The leading cause of tonsil hypertrophy in children is viruses, often in combination with bacterial flora. The purpose — to evaluate the role of the main viral-bacterial pathogens in pharyngeal tonsil hypertrophy (PTH) and adenoiditis in children. Material and methods. We observed 463 children with PTH and chronic adenoiditis. The patients were divided into groups depending on the PTH degree: 108 children with grade 1–2 adenoids, 355 children with PTH grade 3. EBV DNA, CMV and HHV-6 were determined in a smear from removed tissue of the pharyngeal tonsil or in a swab from the nasopharynx using the PCR method, and a bacteriological examination of a smear from the throat and nose was performed. The sIgA proteinase activity of the isolated S. aureus strains was determined. The content of sIgA in the patients’ saliva was determined. Statistical processing of data was carried out using Statistica 8.0 software. Results. DNA of herpes viruses in swabs from the nasopharynx was detected in 70.8% (328/463) of the examined: in 36.1% (39/108) of patients with grade 1–2 PTH and 81.4% (289/355) of grade 3, p < 0.001. Epstein-Barr virus was isolated most often – in 53% (174/328). The results of a bacteriological study of 151 patients with PTH demonstrated the predominance of S. aureus (71.5%, 108/151) and S. pyogenes (55.6%, 84/151), p = 0.005 in nasopharyngeal swabs. 58.3% (63/108) of S. aureus strains had sIgA proteinase activity. In 45.3% (49/108) of cases, the isolation of such staphylococci was combined with the detection of EBV DNA. The level of sIgA in the saliva of patients with grade 3 tonsil hypertrophy (Me 43; IQR 34.4–76.6) was statistically significantly lower than the level of sIgA in the saliva of patients with grade 1–2 hypertrophy of the tonsils (Me 63.8; IQR 44.2–98,6), p = 0.023 and healthy children. Conclusions. Infection of the nasopharynx with EBV can be considered a leading risk factor for the development of tonsil hypertrophy and adenoiditis in children. This must be taken into account when monitoring such patients.

Перспективы использования иммуностимулирующих средств с неспецифической противовирусной активностью в комплексном лечении детей с хроническим аденоидитом

The article presents the results of studies on using inosine pranobex in children with pharyngeal tonsils hypertrophy of the 3rd degree after surgical intervention to prevent the development of pharyngeal tonsillitis relapses and compensatory hypertrophy of palatine tonsils in the postoperative period. 355 children aged 3 to 7 years old with a diagnosis of pharyngeal tonsil hypertrophy of the 3rd degree were examined. Laboratory methods of examination included qualitative studies to detect DNA of lymphotropic herpesviruses (EBV, CMV, HHV-6) in flushes from the nasopharyngeal mucosa. As a result of the conducted research, it was revealed that the inclusion of inosine pranobex in the complex treatment of children with chronic adenoiditis with verified herpesvirus infection is clinically effective and helps to reduce the risk of recurrence of pharyngeal tonsil hypertrophy and compensatory palatine tonsil hypertrophy.

Герпесвирусные инфекции в формировании острой и хронической патологии: обоснование необходимости своевременной терапии

Objective. To determine the frequency and possible clinical significance of herpesvirus DNA detection in various organs. Materials and methods. The study included 168 samples of the cerebral cortex, myocardium, vascular walls, liver, kidneys, and lungs from 28 deceased people. The causes of death were cardiopulmonary insufficiency due to decompensation of atherosclerosis (15 (54%) patients, study group) or intoxication due to septic conditions (13 (46%) patients, comparison group). DNA viruses were detected using the polymerase chain reaction. Results. The most frequently detected virus was human herpes virus type 6 (HHV-6) (82%), less frequently – Epstein-Barr virus (58%), cytomegalovirus and herpes simplex virus type 1 (17% each). In the study group, HHV-6 DNA was more often found in the myocardium (87%), cerebral cortex (47%) and liver (40%). Conclusion. In 87% of cases of death from decompensation of atherosclerotic changes, HHV-6 DNA was found in altered areas of the myocardium. In the brain, liver, kidneys and lungs, HHV-6 was detected less frequently. The association of the presence of HHV-6 DNA with the severity of changes in the heart, brain, liver and kidneys and the presence of other herpesviruses with symptoms of acute respiratory viral infection require the prescription of an antiviral agent with a broad spectrum of action. Inosine pranobex is indicated to prevent the progression of somatic pathology by stopping the activity of herpesvirus infection, which often accompanies or is the main factor in the symptoms of acute respiratory viral infection. Key words: HHV-6, atherosclerosis, human herpes virus type 6, herpes, herpesvirus infection, ARVI, PCR, inosine pranobex

Prevalence of Herpesvirus DNA in Corneal Transplant Recipients

Purpose: Graft failure after penetrating keratoplasty (PK) is a serious complication, especially in eyes with herpetic keratitis (HK). This study evaluated the prevalence and graft survival of herpes simplex virus type 1 (HSV-1) and varicella zoster virus (VZV) DNA in recipient corneas during PK. Methods: The retrospective study was performed at the Department of Ophthalmology at University Hospital in Mainz, Germany. We analyzed data from every patient who underwent PK between January 2020 and June 2021. According to our clinical routine, we performed HSV-1 and VZV polymerase chain reaction (PCR) on all excised corneal buttons regardless of the primary clinical diagnosis. Results: We included 112 eyes of 112 consecutive patients who underwent PK. At the time of PK, 91 (81.25%) patients had no history of HK and 21 (18.75%) patients did. The recipient corneas of 91 patients without a history of HK tested positive for HSV-1 DNA in 12 (13.2%) eyes, for VZV DNA in 3 (3.3%) eyes, and for HSV-1 and VZV DNA simultaneously in 2 (2.2%) eyes. The recipient corneas of 21 patients with a preoperative history of HK tested positive for HSV-1 DNA in 13 (61.9%) eyes and VZV DNA in 1 (4.8%) eye. All patients with positive herpes DNA and no history of HK prior to PK received antiherpetic treatment and had a 100% graft survival rate after 1 year. Conclusions: We found herpesvirus DNA in 18.7% of recipient corneas without clinical suspicion or history of herpes keratitis. This suggests the need of routine HSV-1 and VZV PCR testing in all explanted corneas regardless of clinical suspicion, to detect, treat and prevent possible recurrence of herpes infection in corneal grafts and support graft survival.

Artificial intelligence in the immunodiagnostics of chronic periodontitis

Artificial intelligence is used to diagnose various diseases of the oral cavity. In the field of clinical laboratory diagnostics, machine learning algorithms are used in the interpretation of complex biochemical data. The purpose of this study was to search for significant infectious-immunological clinical and laboratory data based on a machine learning algorithm for chronic periodontitis. To do this, 124 patients aged 40 to 70 years diagnosed with chronic periodontitis were examined by real-time PCR to detect the periodontal pocket DNA of human herpes viruses and bacterial periodontopathogenic microflora Fusobacterium nucleatum, Treponema denticola, Porphyromonas endodontalis etc., and Porphyromonas gingivalis. Matrix RNAs of proinflammatory cytokines and other markers of chronic inflammatory process were also studied: IL-1, IL-10, IL-18, TNFa, TLR4, GATA3, CD68. TNFa, IFNg, IL-1, IL-4, IL-6, IL-10, IL-18; VEGF were determined in a dentoalveolar fluid. Immune cells of the oral cavity were evaluated by analyzing level of CD3+, CD4+, CD8+, CD3+HLA-DR+, CD64+16+14, CD4+25+127+low, CD3+CD16+CD56+, CD3CD16+CD56+, CD14+, CD14+HLA-DR+, CD19+HLA-DR+, CD19+CD5+B27, CD19+CD5B27, CD19+CD5B27+ cells. Random forest machine learning was used to evaluate the data. A relationship between pathogenic microflora and modality of immune response was revealed. The proinflammatory component reflected in the expression of IL-1, TNFa, and IFNg mRNA, prevailed in the immune response against aggressive periodontal pathogens: T. denticola, F. nucleatum, etc. The random forest machine learning algorithm selected correlation ratios r 0.5 (both positive and negative) from a set of data for further analysis by the operator. The random forest machine learning model showed the following significant combinations of data by 10% with a teacher: VEGF, CD3+, CD14+HLA-DR, CD19+CD5CD27+, as well as TLR4, IL-1b, IL-10, TNFa, and IL-18 mRNA. The development of the applied random forest machine learning model with a teacher has already shown a 25% difference: P. endodontalis, GATA3, CD3+, CD14+, CD19+CD5CD27+, as well as TLR4, TNFa, IL-1b, IL-10, and IL-18 mRNA. The search for significant infectious-immunological clinical and laboratory data based on a machine learning algorithm for chronic periodontitis has shown the importance of proinflammatory cytokines, monocytes, T-lymphocytes and memory B-cells in the development of osteodestructive inflammatory process of mRNA to reveal non-evident causality factors.

Analysis of herpesvirus infection and genome single nucleotide polymorphism risk factors in multiple sclerosis, Volga federal district, Russia.

Multiple sclerosis (MS) is a heterogeneous disease where herpesvirus infection and genetic predisposition are identified as the most consistent risk factors. Serum and blood samples were collected from 151 MS and 70 controls and used to analyze circulating antibodies for, and DNA of, Epstein Barr virus (EBV), human cytomegalovirus (HCMV), human herpes virus 6 (HHV6), and varicella zoster virus (VZV). The frequency of selected single nucleotide polymorphisms (SNPs) in MS and controls were studied. Herpesvirus DNA in blood samples were analyzed using qPCR. Anti-herpesvirus antibodies were detected by ELISA. SNPs were analyzed by the allele-specific PCR. For statistical analysis, Fisher exact test, odds ratio and Kruskall-Wallis test were used; p<0.05 values were considered as significant. We have found an association between circulating anti-HHV6 antibodies and MS diagnosis. We also confirmed higher frequency of A and C alleles in rs2300747 and rs12044852 of CD58 gene and G allele in rs929230 of CD6 gene in MS as compared to controls. Fatigue symptom was linked to AC and AA genotype in rs12044852 of CD58 gene. An interesting observation was finding higher frequency of GG genotype in rs12722489 of IL2RA and T allele in rs1535045 of CD40 genes in patient having anti-HHV6 antibodies. A link was found between having anti-VZV antibodies in MS and CC genotype in rs1883832 of CD40 gene.

Screening of West Nile Virus, Herpesvirus, and Parvovirus in Rattus spp. in Klang Valley, Malaysia

Free-roaming and scavenging lifestyles of Rattus spp. in densely populated urban areas expose them to multiple pathogens that facilitate the transmission of infection to the human population more rapidly, raising public health concerns. There is limited information on the status of rat susceptibility to virus infection, particularly West Nile virus (WNV), herpesvirus, and parvovirus, to prepare for emerging zoonosis. A total of 23 (n = 23) blood samples collected from Rattus spp. in the wet market areas of Klang Valley, Malaysia, were subjected to molecular assay using a one-step reverse transcription-polymerase chain reaction (RT-PCR) to detect the highly conserved region of the WNV capsid and pre-membrane protein via nested polymerase chain reaction (PCR) assay targeting highly conserved amino acid motifs within the herpesviral DNA-directed DNA polymerase gene (DPOL) and polymerase chain reaction (PCR) assay targeting the parvovirus non-structural (NS) protein. As a result, 4 out of 23 (17.39%) rats were positive for herpesvirus DNA, but none were positive for WNV RNA and parvovirus DNA. The positive PCR amplicons of herpesvirus DNA were subjected to partial DNA sequencing analysis, 100% identical to Acomys herpesvirus SVMS 226,222 from Betaherpesvirinae, which is highly suggestive of rat cytomegalovirus (RCMV). This study has successfully demonstrated the presence of RCMV from Rattus spp. in the Klang Valley. The RCMV potentially crosses species barriers and establishes infection, raising public health concerns. The non-viraemic state of WNV or parvovirus infection, low sample size, and limited niche distribution emphasise the need for the expansion of this study in the future.

Herpesvirus and adenovirus surveillance in threatened wild West Indian (Trichechus manatus) and Amazonian manatees (Trichechus inunguis), Brazil

The family Trichechidae (order Sirenia) comprises three speciesAfrican (Trichechus senegalenses), West Indian (T. manatus), WIM)], and the Amazonian manatees (T. inunguis, AMM). Whereas WIM inhabits both riverine and coastal systems in the western Atlantic, AMM is the only exclusively freshwater sirenian, endemic to the Amazon River Basin. The study of infectious agents is essential to species conservation, especially considering that both species are classified as Vulnerable by the IUCN Red List and as Endangered by the Brazilian Red List. The current knowledge about viral agents in sirenians is scarce. Herpesviruses and adenovirus are DNA viruses able to infect and cause disease in a wide range of hosts. Herein, we used panPCR protocols to survey herpesvirus and adenovirus in blood samples of wild WIM (n = 23) and AMM (n = 26) under human care in Brazil. Herpesvirus DNA was detected in one juvenile female WIM (1/23; 4.3%; 95% CI -4.7 – 13.3) from Ceará state and in four AMM (two juvenile females, a juvenile male, and an adult female; 4/26; 15.4%; 95% CI 0.5 – 30.3) from Amazonas state. The two different gammaherpesvirus DNA polymerase sequence types identified (one per species, a sequence type in a WIM and another one in three AMM) were highly similar (99% nucleotide identity) to Trichechid herpesvirus 1, reported in West Indian manatees of Florida (USA), and 100% identical when translated into amino acids. A herpesviral glycoprotein B sequence was identified in two AMM. None of the samples was positive to adenovirus. To the authors’ knowledge, this is the first herpesvirus detection in manatees from South America, expanding the herpesvirus geographical range, and the first in WIM and AMM worldwide. Our findings suggest (i) that West Indian and Amazonian manatees are possibly the natural hosts of the detected herpesvirus, and (ii) coevolution of that gammaherpesvirus with Trichechus. Future studies are necessary to characterize the obtained virus and elucidate potential pathological effects (if any) in these species.

Retraction: BRCA1 Regulates IFI16 Mediated Nuclear Innate Sensing of Herpes Viral DNA and Subsequent Induction of the Innate Inflammasome and Interferon-β Responses.

Retraction: BRCA1 Regulates IFI16 Mediated Nuclear Innate Sensing of Herpes Viral DNA and Subsequent Induction of the Innate Inflammasome and Interferon-β Responses.

Nucleotide tetramers TCGA and CTAG: viral DNA and the genetic code (hypothesis)

Introduction. The published and our own data show that CTAG and, to a lesser extent, TCGA tetra-nucleotides have significantly lower concentrations in frequency profiles (FPs) of herpesvirus DNAs compared to other complete, bilaterally symmetrical tetra-nucleotides.The aim of the study is to present a comparative analysis of CTAG and TCGA tetra-nucleotide FPs in viral DNAs.Materials and methods. We have analyzed FPs and other characteristics of the two above tetramers in DNAs of at least one species of viruses of each genus (or each subfamily, if the classification into genera was not available), complying with the size limit requirements (minimum 100,000 base pairs) — a total of more than 200 species of viruses. The analysis was performed using the GenBank database.Results. Two groups of characteristics of TCGA and CTAG tetramers have been described. One of them covers the results of the FP analysis for these tetranucleotides in viral DNAs and shows that DNAs with GC:AT 2 are characterized by nCGn FP symmetries while these symmetries are frequently distorted in nTAn FP due to CTAG underrepresentation. The other group of tetramer characteristics demonstrates differences in their FPs in complete viral DNAs and in their genomes (a coding part, which can reach 80% in some studied viruses, thus making the analysis of their DNAs more significant than the analysis of DNAs of cellular live forms) and suggests that these tetramers may have participated in the origin of the universal genetic code.Discussion. Assumedly, the genetic code started evolving amid C+G prevailing in "pre-code" DNA polymers; then the initial code forms evolved further to their final structure where TCGA and CTAG tetramers hold a central position, encapsulating the previous stages of this evolution. The nCGn FP symmetries typical of the "complete" DNA of Herpes simplex viruses disappear in the sequence of the second codon letters of the genome of these viruses, implying that their functions differ from functions of other letters and emphasizing the reasonableness of presenting the genetic code as a calligram where the second line is not symmetrical.

Multiplex real-time PCR for the detection and differentiation of equid gammaherpesvirus 2 and 5

Equid gammaherpesvirus 2 (EHV-2) and 5 (EHV-5) are widely distributed in the equines. Although their pathogenic potential is not yet fully understood, they appear to play a role in disease patterns like equine multinodular pulmonary fibrosis. In this study, a multiplex real-time PCR (rtPCR) was designed to detect DNA of the glycoprotein H (EHV-2) and E11 gene (EHV-5). Analytical specificity was determined by testing DNA of other herpesviruses by SYBR Green rtPCR and melting curve analysis, as well as Sanger sequencing of positive field samples. Analytical sensitivity was assessed by standard curve generation of serial plasmid dilutions containing the respective target gene. Melting curves and BLAST analysis of the sequences indicated specific detection of the viruses. The lower limit of detection of the singleplex rtPCR was 40 and 29 DNA copies per reaction for EHV-2 and EHV-5, respectively. Comparison of the Ct values of a selection of positive field samples showed only minimal differences between the singleplex and the multiplex assay. The here described multiplex rtPCR protocol allows sensitive and specific detection of EHV-2 and EHV-5. It represents a convenient and rapid tool for future studies to investigate the clinical relevance of EHV-2 and EHV-5 in more detail.

Specific inhibition of the interaction between pseudorabies virus DNA polymerase subunits UL30 and UL42 by a synthetic peptide

Pseudorabies virus (PRV) is a ubiquitous and economically important swine alphaherpesvirus that causes devastating swine diseases worldwide. PRV-encoded DNA-dependent DNA polymerase, comprised of the catalytic subunit UL30 and the accessory subunit UL42, is essential for viral replication. PRV UL30 and UL42 act as a heterodimer with UL30 harboring inherent DNA polymerase activity and UL42 conferring processivity on the DNA polymerase holoenzyme. The formation of PRV UL30/UL42 heterodimer holoenzyme through protein-protein interactions is indispensable for viral replication. In work described here, we defined the key domains that mediate PRV UL30/UL42 interaction, and found that the 41 carboxy-terminal amino acids region of PRV UL30 is critical for its interaction with UL42. Intriguingly, a synthetic peptide corresponding to these 41 carboxy-terminal amino acid residues efficiently disrupted PRV UL30/UL42 interaction through competitively binding to UL42. These findings suggest that the peptides from the PRV DNA polymerase UL30/UL42 subunit interface may represent potential targets for designing a novel intervention strategy against PRV infection. This work further strengthens the concept that the herpesvirus DNA polymerase catalytic subunits utilize their extreme carboxy-terminal domains as a conserved mechanism to associate with their cognate accessory subunits, providing us the opportunity of designing novel antiviral agents against herpesvirus infection through disruption of the herpesvirus DNA polymerase subunit interactions.

Detection of herpesviral hematopoietic necrosis virus (Cyprinid herpesvirus 2, CyHV-2) from goldfish, Carassius auratus (L.) in Korea

In April and June 2014, mortalities of goldfish occurred at Korea. The principal signs included pale gills, severely enlarged and softened spleen and kidney and red liver. Moreover, the histopathological characteristics observed mainly in hematopoietic tissues of kidney, gill lamellae and intestine. The predominant histopathological changes were severe necrosis of hematopoietic tissue. In addition, nucleus exhibiting peripherally displaced chromatin were particularly abundant in the kidney of affected fish. The histological examination led to hypothesize the implication of a virus in the mortality. The hematopoietic necrosis herpes-virus (Cyprinid herpesvirus 2, CyHV-2) DNA polymerase gene successfully detected in DNA extracted from kidney and spleen of affected fish using PCR assay and showed 100% identity with already deposited CyHV-2 DNA polymerase gene in NCBI. Artificial infection trials using affected tissues filtrates gave cumulative mortalities of 30% for virus injected goldfish. In the present study, CyHV-2 was detected and identified as the causative pathogen of the epizootic in goldfish in Korea.