The infection of Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the main causes of economic losses in sericulture. Thus, it is essential to establish rapid and effective method for BmNPV detection. In the present study, we have developed a recombinase-aided amplification (RAA) to amplify the BmNPV genomic DNA at 37 °C within 30 min, and achieved a rapid detection method by coupling with a lateral flow dipstick (LFD). The RAA-LFD method had a satisfactory detection limit of 6 copies/μL of recombinant plasmid pMD19-T-IE1, and BmNPV infection of silkworm can be detected 12 h post-infection. This method was highly specific for BmNPV, and without cross-reactivity to other silkworm pathogens. In contrast to conventional polymerase chain reaction (PCR), the RAA-LFD assay showed higher sensitivity, cost-saving, and especially is apt to on-site detection of BmNPV infection in the sericulture production.
- MeSH
- bourec * virologie MeSH
- DNA virů genetika MeSH
- limita detekce MeSH
- nukleopolyhedroviry * genetika izolace a purifikace MeSH
- rekombinasy * metabolismus genetika MeSH
- senzitivita a specificita MeSH
- techniky amplifikace nukleových kyselin * metody MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
Cíl: Přehledová práce poskytuje základní praktické informace o využití biosenzorů v rychlé diagnostice virových patogenů. Výsledky: Viry díky evolučním změnám v genomu přeskakují do lidské populace, u níž vyvolávají závažné epidemie. Rychlé diagnostické nástroje POCT založené na biosenzorech umožní jejich rozpoznání pro potřeby klinické diagnostiky mimo specializované laboratoře. Kombinace těchto zařízení s technikami 3D tisku, mikrofluidních systémů, nanotechnologie a elektrochemické detekce výrazně zvyšuje využitelnost biosenzorů v laboratorní medicíně. Intenzivní nanomedicínský výzkum probíhá u celé řady virů, např. HIV, ebola, chřipka a viry hepatitid. V souvislosti s celosvětovou pandemií covid-19 se v současné době vývoj nanobiosenzorů ubírá především směrem k detekci SARS-CoV-2. Závěr: Dostupná literární data naznačují, že rychlé senzory a biosenzory mají značný klinický potenciál pro využití v POCT.
Aim: The review provides basic practical information about the use of biosensors in the rapid diagnosis of viral pathogens. Results: Thanks to evolutionary changes in the genome, viruses jump into the human population, where they cause serious epidemics. Rapid POCT diagnostic tools based on biosensors will enable their use for clinical diagnosis needs outside of specialized laboratories. The combination of these devices with the techniques of 3D printing, microfluidic systems, nanotechnology and electrochemical detection significantly increases the usability of biosensors. Intensive research is carried out on a wide range of viruses, e.g. HIV, Ebola, influenza, hepatitis viruses. In connection with the global covid-19 pandemic, the development of nanobiosensors is currently focused primarily on the detection of SARS-CoV-2. Conclusion: Available literature data suggest that fast sensors and biosensors have considerable clinical potential for the use in POCT.
- MeSH
- biosenzitivní techniky * MeSH
- DNA virů analýza MeSH
- elektrochemie MeSH
- lidé MeSH
- nanomedicína MeSH
- nanotechnologie MeSH
- point of care testing MeSH
- viry * izolace a purifikace MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- přehledy MeSH
DNA virus infections are often lifelong and can cause serious diseases in their hosts. Their recognition by the sensors of the innate immune system represents the front line of host defence. Understanding the molecular mechanisms of innate immunity responses is an important prerequisite for the design of effective antivirotics. This review focuses on the present state of knowledge surrounding the mechanisms of viral DNA genome sensing and the main induced pathways of innate immunity responses. The studies that have been performed to date indicate that herpesviruses, adenoviruses, and polyomaviruses are sensed by various DNA sensors. In non-immune cells, STING pathways have been shown to be activated by cGAS, IFI16, DDX41, or DNA-PK. The activation of TLR9 has mainly been described in pDCs and in other immune cells. Importantly, studies on herpesviruses have unveiled novel participants (BRCA1, H2B, or DNA-PK) in the IFI16 sensing pathway. Polyomavirus studies have revealed that, in addition to viral DNA, micronuclei are released into the cytosol due to genotoxic stress. Papillomaviruses, HBV, and HIV have been shown to evade DNA sensing by sophisticated intracellular trafficking, unique cell tropism, and viral or cellular protein actions that prevent or block DNA sensing. Further research is required to fully understand the interplay between viruses and DNA sensors.
- MeSH
- DNA virů metabolismus MeSH
- Herpesviridae * genetika metabolismus MeSH
- infekce DNA virem * MeSH
- lidé MeSH
- Polyomavirus * genetika MeSH
- přirozená imunita MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
Anelloviruses are small negative-sense single-stranded DNA viruses with genomes ranging in size from 1.6 to 3.9 kb. The family Anelloviridae comprised 14 genera before the present changes. However, in the last five years, a large number of diverse anelloviruses have been identified in various organisms. Here, we undertake a global analysis of mammalian anelloviruses whose full genome sequences have been determined and have an intact open reading frame 1 (ORF1). We established new criteria for the classification of anelloviruses, and, based on our analyses, we establish new genera and species to accommodate the unclassified anelloviruses. We also note that based on the updated species demarcation criteria, some previously assigned species (n = 10) merge with other species. Given the rate at which virus sequence data are accumulating, and with the identification of diverse anelloviruses, we acknowledge that the taxonomy will have to be dynamic and continuously evolve to accommodate new members.
- MeSH
- Anelloviridae klasifikace genetika MeSH
- databáze genetické MeSH
- DNA virů genetika MeSH
- fylogeneze MeSH
- genom virový genetika MeSH
- otevřené čtecí rámce genetika MeSH
- savci virologie MeSH
- sekvence nukleotidů MeSH
- terminologie jako téma MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The genus Gyrovirus was assigned to the family Anelloviridae in 2017 with only one recognized species, Chicken anemia virus. Over the last decade, many diverse viruses related to chicken anemia virus have been identified but not classified. Here, we provide a framework for the classification of new species in the genus Gyrovirus and communicate the establishment of nine new species. We adopted the 'Genus + freeform epithet' binomial system for the naming of these species.
- MeSH
- Anelloviridae klasifikace genetika MeSH
- databáze genetické MeSH
- DNA virů genetika MeSH
- fylogeneze MeSH
- genom virový genetika MeSH
- Gyrovirus klasifikace genetika MeSH
- lidé MeSH
- sekvenční analýza DNA MeSH
- terminologie jako téma * MeSH
- virové plášťové proteiny genetika MeSH
- virus anemie kuřat klasifikace genetika MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Anal cancer (AC) screening is justified in high-risk populations, particularly HIV-positive men having sex with men (MSM). HR-HPV testing could improve the efficiency of cytologically based screening of AC, as in the screening of biologically analogical cervical cancer. The specificity of HR-HPV testing is influenced by the prevalence of HR-HPV infection in the screened population. Reported anal HR-HPV DNA prevalence in MSM is high, but HR-HPV mRNA reflects rather long-term infections and is more specific for high-grade lesions. However, no data were published about HR-HPV DNA and mRNA prevalence in the Czech AC screening population. METHOD: Results of liquid-based anal cytology of 203 predominantly HIV-positive MSM from the Czech AC screening cohort were correlated with results of DNA and E6/E7 mRNA testing of 14 HR-HPV types, and HPV16 genotyping. Eighty-one MSM underwent a standard anoscopy. RESULTS: A total of 109 (53.7%) samples had abnormal cytology, with 12 (5.9%) ASC-H/HSIL, 67 (33.0%) samples cytologically negative, and 27 (13.3%) unsatisfactory. HR-HPV DNA was detected in 134 (66.0%) and HR-HPV RNA in 72 (35.5%) anal smears. HR-HPV mRNA and HPV16 mRNA positivity were associated with abnormal cytology (p = .0037, p = .0021). No significant association was found between HR-HPV DNA or HPV16 DNA positivity and abnormal cytology. No high-grade lesions were revealed by anoscopy. CONCLUSION: Prevalence of anal HR-HPV DNA among Czech MSM is high, however, the prevalence of HR-HPV mRNA is half and associated with abnormal cytology. Our results indicate an increased efficiency of cytological screening when combined with HR-HPV mRNA testing.
- MeSH
- časná detekce nádoru statistika a číselné údaje MeSH
- DNA virů genetika MeSH
- dospělí MeSH
- homosexualita mužská genetika statistika a číselné údaje MeSH
- infekce papilomavirem epidemiologie patologie virologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- messenger RNA genetika MeSH
- mladý dospělý MeSH
- nádory anu genetika patologie MeSH
- prevalence MeSH
- sexuální a genderové menšiny statistika a číselné údaje MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
The mechanism by which DNA viruses interact with different DNA sensors and their connection with the activation of interferon (IFN) type I pathway are poorly understood. We investigated the roles of protein 204 (p204) and cyclic guanosine-adenosine synthetase (cGAS) sensors during infection with mouse polyomavirus (MPyV). The phosphorylation of IFN regulatory factor 3 (IRF3) and the stimulator of IFN genes (STING) proteins and the upregulation of IFN beta (IFN-β) and MX Dynamin Like GTPase 1 (MX-1) genes were detected at the time of replication of MPyV genomes in the nucleus. STING knockout abolished the IFN response. Infection with a mutant virus that exhibits defective nuclear entry via nucleopores and that accumulates in the cytoplasm confirmed that replication of viral genomes in the nucleus is required for IFN induction. The importance of both DNA sensors, p204 and cGAS, in MPyV-induced IFN response was demonstrated by downregulation of the IFN pathway observed in p204-knockdown and cGAS-knockout cells. Confocal microscopy revealed the colocalization of p204 with MPyV genomes in the nucleus. cGAS was found in the cytoplasm, colocalizing with viral DNA leaked from the nucleus and with DNA within micronucleus-like bodies, but also with the MPyV genomes in the nucleus. However, 2'3'-Cyclic guanosine monophosphate-adenosine monophosphate synthesized by cGAS was detected exclusively in the cytoplasm. Biochemical assays revealed no evidence of functional interaction between cGAS and p204 in the nucleus. Our results provide evidence for the complex interactions of MPyV and DNA sensors including the sensing of viral genomes in the nucleus by p204 and of leaked viral DNA and micronucleus-like bodies in the cytoplasm by cGAS.
- MeSH
- DNA virů genetika imunologie MeSH
- fosfoproteiny antagonisté a inhibitory genetika metabolismus MeSH
- fosforylace MeSH
- infekce onkogenními viry imunologie virologie MeSH
- interakce hostitele a patogenu MeSH
- interferon beta metabolismus MeSH
- jaderné proteiny antagonisté a inhibitory genetika metabolismus MeSH
- membránové proteiny antagonisté a inhibitory genetika metabolismus MeSH
- myši MeSH
- nukleotidyltransferasy antagonisté a inhibitory genetika metabolismus MeSH
- polyomavirové infekce imunologie virologie MeSH
- Polyomavirus genetika imunologie MeSH
- přirozená imunita imunologie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
G-quadruplexes contribute to the regulation of key molecular processes. Their utilization for antiviral therapy is an emerging field of contemporary research. Here we present comprehensive analyses of the presence and localization of putative G-quadruplex forming sequences (PQS) in all viral genomes currently available in the NCBI database (including subviral agents). The G4Hunter algorithm was applied to a pool of 11,000 accessible viral genomes representing 350 Mbp in total. PQS frequencies differ across evolutionary groups of viruses, and are enriched in repeats, replication origins, 5'UTRs and 3'UTRs. Importantly, PQS presence and localization is connected to viral lifecycles and corresponds to the type of viral infection rather than to nucleic acid type; while viruses routinely causing persistent infections in Metazoa hosts are enriched for PQS, viruses causing acute infections are significantly depleted for PQS. The unique localization of PQS identifies the importance of G-quadruplex-based regulation of viral replication and life cycle, providing a tool for potential therapeutic targeting.
- MeSH
- databáze nukleových kyselin * MeSH
- DNA virů genetika metabolismus MeSH
- G-kvadruplexy * MeSH
- genom virový * MeSH
- lidé MeSH
- virové nemoci * genetika metabolismus MeSH
- viry * genetika metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
