Novel Resistance-Associated Mutations of Thymidine Kinase and Dna Polymerase Genes of Herpes Simplex Virus Type 1 and Type 2

Sauerbrei, Andreas; Bohn, Kristin; Heim, Albert; Hofmann, Jörg; Weißbrich, Benedikt; Schnitzler, Paul; Hoffmann, Dieter; Zell, Roland; Jahn, Gerhard; Wutzler, Peter; Hamprecht, Klaus

doi:10.3851/imp1870

Cited by 57 publications

(47 citation statements)

References 35 publications

(65 reference statements)

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“…Interestingly, we were able to recover in one case (sample 1,538) Ͼ56% of the HSV-1 genome, which allowed us to analyze the potential resistance of this virus against acyclovir. Moreover, the UL23 open reading frame, encoding thymidine kinase (TK), showed 100% sequence identity (data not shown) to an HSV-1 strain recently recovered from a bone marrow transplant patient which was clinically resistant against acyclovir treatment (43).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Unbiased Next-Generation Sequencing of RNA (RNA-seq) as a Diagnostic Method in Influenza Virus-Positive Respiratory Samples

et al. 2015

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e Unbiased nontargeted metagenomic RNA sequencing (UMERS) has the advantage to detect known as well as unknown pathogens and, thus, can significantly improve the detection of viral, bacterial, parasitic, and fungal sequences in public health settings. In particular, conventional diagnostic methods successfully identify the putative pathogenic agent in only 30% to 40% of respiratory specimens from patients with acute respiratory illness. Here, we applied UMERS to 24 diagnostic respiratory specimens (bronchoalveolar lavage [BAL] fluid, sputum samples, and a swab) from patients with seasonal influenza infection and 5 BAL fluid samples from patients with pneumonia that tested negative for influenza to validate RNA sequencing as an unbiased diagnostic tool in comparison to conventional diagnostic methods. In addition to our comparison to PCR, we evaluated the potential to retrieve comprehensive influenza virus genomic information and the capability to detect known superinfecting pathogens. Compared to quantitative real-time PCR for influenza viral sequences, UMERS detected influenza viral sequences in 18 of 24 samples. Complete influenza virus genomes could be assembled from 8 samples. Furthermore, in 3 of 24 influenza-positive samples, additional viral pathogens could be detected, and 2 of 24 samples showed a significantly increased abundance of individual bacterial species known to cause superinfections during an influenza virus infection. Thus, analysis of respiratory samples from known or suspected influenza patients by UMERS provides valuable information that is relevant for clinical investigation. Influenza has a severe impact on our health system, not only owing to its potential to cause worldwide pandemics but also due to the high number of seasonal infections. Bacterial and/or viral coinfections and subsequent pneumonia can lead to enhanced illness in elderly and immunosuppressed patients. Approximately 0.5% of all influenza A infections in healthy younger adults and 2.5% of influenza A infections in the elderly and younger children are accompanied by severe bacterial-induced pneumonia (1, 2). These numbers are significantly higher during pandemic episodes (3-5). The most common causes of coinfections observed in both pandemic and seasonal episodes of influenza A infections are Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus influenzae, and Streptococcus pyogenes (6, 7). This relationship between influenza virus and bacterial pathogenicity is underlined by several studies using animal models. For example, mice infected with influenza virus or Streptococcus pneumoniae alone showed mortality rates of 35% and 15%, respectively, whereas mice coinfected with influenza virus and Streptococcus pneumoniae displayed a 100% mortality rate (8). Furthermore, several studies in humans indicate that colonization with Streptococcus pneumoniae increases the risk of severe complications associated with influenza A viral infection (9, 10), thus highlighting the importance of rapid diagnosis of bacterial coinfecti...

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Section: Discussionmentioning

confidence: 99%

Evaluation of Unbiased Next-Generation Sequencing of RNA (RNA-seq) as a Diagnostic Method in Influenza Virus-Positive Respiratory Samples

et al. 2015

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“…Additionally, the known (16) but so far unclear substitution A118V could be characterized as a natural polymorphism (Table 1). Furthermore, the amino acid change A156V, which was reported previously as being associated with ACV resistance (16), was detected in one strain (no. 185-93) with an ACV-sensitive phenotype.…”

Section: Genotypic Resistance Testing (I) Thymidine Kinasementioning

confidence: 99%

“…Only isolates with novel or unclear nonsynonymous mutations in the DNA Pol or ACV resistance-associated genotype were tested against FOS (AstraZeneca, Wilmslow, United Kingdom). All phenotypic analyses were performed using a plaque reduction assay, including the formazan test according to the method described previously (16,17,19,21). In short, Vero 76 cells, only used for testing FOS, or human Caucasian fetal lung fibroblasts of the cell line Wi 38 (European Collection of Cell Cultures, Salisbury, United Kingdom), exclusively used for analyzing the phenotype to ACV, were seeded at a density of 10 5 ml Ϫ1 .…”

Section: Methodsmentioning

confidence: 99%

“…Nonetheless, several mutations have been shown to confer resistance despite their location outside conserved gene regions (16). The identification of these mutations is complicated by the high gene variability of TK and DNA Pol (16)(17)(18)(19) as well as the complexity and intricacy of drug resistance (2,20) due to the increasing spectrum of novel mutations in both genes. Thus, a continuous determination of both genotypes is required.…”

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confidence: 99%

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Sequence Analysis of Herpes Simplex Virus 1 Thymidine Kinase and DNA Polymerase Genes from over 300 Clinical Isolates from 1973 to 2014 Finds Novel Mutations That May Be Relevant for Development of Antiviral Resistance

Schmidt

Bohn-Wippert

Zell

et al. 2015

Antimicrob Agents Chemother

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A total of 302 clinical herpes simplex virus 1 (HSV-1) strains, collected over 4 decades from 1973 to 2014, were characterized retrospectively for drug resistance. All HSV-1 isolates were analyzed genotypically for nonsynonymous mutations in the thymidine kinase (TK) and DNA polymerase (Pol) genes. The resistance phenotype against acyclovir (ACV) and/or foscarnet (FOS) was examined in the case of novel, unclear, or resistance-related mutations. Twenty-six novel natural polymorphisms could be detected in the TK gene and 69 in the DNA Pol gene. Furthermore, three novel resistance-associated mutations (two in the TK gene and one in the DNA Pol gene) were analyzed, and eight known but hitherto unclear amino acid substitutions (two encoded in TK and six in the DNA Pol gene) could be clarified. Between 1973 and 2014, the distribution of amino acid changes related to the natural gene polymorphisms of TK and DNA Pol remained largely stable. Resistance to ACV was confirmed phenotypically for 16 isolates, and resistance to ACV plus FOS was confirmed for 1 isolate. Acyclovir-resistant strains were observed from the year 1995 onwards, predominantly in immunosuppressed patients, especially those with stem cell transplantation, and the number of ACV-resistant strains increased during the last 2 decades. The data confirm the strong genetic variability among HIV-1 isolates, which is more pronounced in the DNA Pol gene than in the TK gene, and will facilitate considerably the rapid genotypic diagnosis of HSV-1 resistance. Herpes simplex virus 1 (HSV-1) is a ubiquitous human pathogen throughout the world. After primary infection occurring mostly during infancy, the virus remains latently for life in sensory local ganglia. Thereafter, HSV-1 may be reactivated and can cause usually self-limiting orolabial lesions in immunocompetent individuals. In contrast, immunocompromised patients often develop chronic diseases associated with painful and widespread exanthema and/or enanthema. Since its marketing in the 1980s, acyclovir (ACV), a guanosine analog, has been the drug of choice for the treatment of HSV infections; its active form is ACV triphosphate (1, 2). Foscarnet (FOS), a pyrophosphate analog, directly inhibits the viral DNA polymerase (Pol) and can be used successfully as an alternative drug in case of ACV resistance.Acyclovir-resistant HSV-1 isolates have a remarkable clinical prevalence, especially in immunocompromised patients. Summarizing published papers of recent years, the prevalence of ACVresistant HSV strains in immunosuppressed patients ranges between 2.5% and 10.9% (3, 4). For immunocompetent patients, studies between 1985 and 1993 showed that ACV resistance in clinical HSV isolates is Ͻ1.0%, very low (5-8). An overall increase in ACV-resistant HSV strains could not be established in a subsequent study (9).The HSV thymidine kinase (TK) protein is responsible for the conversion of ACV to its active form, and 95% of the mutations encoding antiviral resistance are found in the TK gene (10). Only a few cases of resistanc...

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“…ACV resistance frequencies of even more than 25% have been reported for patients in the allogeneic stem cell or bone marrow transplantation setting (8,9). Because resistance to ACV is mostly acquired through mutations in the HSV thymidine kinase (TK) gene, cross-resistance toward other viral TK-dependent nucleoside analogs, such as penciclovir and famciclovir, is not uncommon (10,11). Although in nucleoside analog-resistant cases TK-independent drugs such as the viral DNA polymerase inhibitors foscarnet and cidofovir can be administered, these compounds frequently mediate severe toxic side effects, particularly in patients with high comorbidities.…”

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confidence: 99%

Overcoming drug-resistant herpes simplex virus (HSV) infection by a humanized antibody

Krawczyk

Arndt

Grosse‐Hovest

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

105

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Despite the availability of antiviral chemotherapy, herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) infections remain a severe global health problem. Of particular concern is the growing incidence of drug resistance in immunocompromised patients, which stresses the urgency to develop new effective treatment alternatives. We have developed a humanized monoclonal antibody (mAb hu2c) that completely abrogates viral cell-to-cell spread, a key mechanism by which HSV-1/2 escapes humoral immune surveillance. Moreover, mAb hu2c neutralized HSV fully independent of complement and/or immune effector cell recruitment in a highly efficient manner. Prophylactic and therapeutic administration of mAb hu2c completely prevented infection-related mortality of severely immunodeficient mice being challenged with a lethal dose of HSV-1. The high neutralization capacity of mAb hu2c was fully maintained toward clinical HSV isolates being multiresistant to standard antiviral drugs, and infection was fully resolved in 7/8 nonobese diabetic/SCID mice being infected with a multidrug resistant HSV-1 patient isolate. Immunohistochemical studies revealed no significant cross-reactivity of the antibody toward human tissues. These features warrant further clinical development of mAb hu2c as an immunotherapeutic compound for the management of severe and particularly drugresistant HSV infections.acyclovir resistance | immunocompromised mice | stem cell transplantation

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Novel Resistance-Associated Mutations of Thymidine Kinase and Dna Polymerase Genes of Herpes Simplex Virus Type 1 and Type 2

Cited by 57 publications

References 35 publications

Evaluation of Unbiased Next-Generation Sequencing of RNA (RNA-seq) as a Diagnostic Method in Influenza Virus-Positive Respiratory Samples

Evaluation of Unbiased Next-Generation Sequencing of RNA (RNA-seq) as a Diagnostic Method in Influenza Virus-Positive Respiratory Samples

Sequence Analysis of Herpes Simplex Virus 1 Thymidine Kinase and DNA Polymerase Genes from over 300 Clinical Isolates from 1973 to 2014 Finds Novel Mutations That May Be Relevant for Development of Antiviral Resistance

Overcoming drug-resistant herpes simplex virus (HSV) infection by a humanized antibody

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