Influenza A viruses are enveloped viruses with a genome of eight single-stranded negative-sense RNA molecules. In virions, RNA segments are found as vRNPs associated with NP proteins. The RdRp enzyme, which catalyzes the replication/transcription of the viral genome, is carried as attached to vRNPs. In this study, it was demonstrated that the PA subunit of the viral RdRp interacts with β-actin proteins by the yeast two-hybrid assay. It was shown that the amino-terminal domains of the β-actin protein bind to the carboxy-terminal moiety of the viral PA protein in the mammalian cells. The results were supported by in silico analysis. Over-expression of the β-actin protein was found to have a negative effect on the viral RdRp activity in mini-replicon, but its mechanism of action has remained unknown. The results suggest that the interaction of β-actin and PA protein, a component of vRNPs, may have a role in the intracellular trafficking of the influenza vRNPs and/or viral transcription.
甲型流感病毒是一种包膜病毒，基因组由八条单链负义 RNA 分子组成。在病毒中，RNA片段以与NP蛋白相关联的vRNP形式存在。催化病毒基因组复制/转录的 RdRp 酶附着在 vRNPs 上。本研究通过酵母双杂交实验证明，病毒 RdRp 的 PA 亚基与 β-actin 蛋白相互作用。在哺乳动物细胞中，β-肌动蛋白的氨基末端与病毒 PA 蛋白的羧基末端结合。这一结果得到了硅学分析的支持。研究发现，β-肌动蛋白的过度表达对迷你复制子中病毒 RdRp 的活性有负面影响，但其作用机制仍不清楚。研究结果表明，β-肌动蛋白与 vRNPs 的组成部分 PA 蛋白的相互作用可能在流感 vRNPs 的细胞内运输和/或病毒转录中发挥作用。
{"title":"The interaction of influenza A virus RNA polymerase PA subunit with the human β-actin protein","authors":"Nazife Gelmez, E. Çağlayan, K. Turan","doi":"10.3389/av.2023.11890","DOIUrl":"https://doi.org/10.3389/av.2023.11890","url":null,"abstract":"Influenza A viruses are enveloped viruses with a genome of eight single-stranded negative-sense RNA molecules. In virions, RNA segments are found as vRNPs associated with NP proteins. The RdRp enzyme, which catalyzes the replication/transcription of the viral genome, is carried as attached to vRNPs. In this study, it was demonstrated that the PA subunit of the viral RdRp interacts with β-actin proteins by the yeast two-hybrid assay. It was shown that the amino-terminal domains of the β-actin protein bind to the carboxy-terminal moiety of the viral PA protein in the mammalian cells. The results were supported by in silico analysis. Over-expression of the β-actin protein was found to have a negative effect on the viral RdRp activity in mini-replicon, but its mechanism of action has remained unknown. The results suggest that the interaction of β-actin and PA protein, a component of vRNPs, may have a role in the intracellular trafficking of the influenza vRNPs and/or viral transcription.","PeriodicalId":7205,"journal":{"name":"Acta virologica","volume":"23 4","pages":""},"PeriodicalIF":1.7,"publicationDate":"2024-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139445490","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Human adenovirus-5 (hAd5) is an important gene delivery vector, which has been widely used in various fields of biomedicine, such as gene therapy, cancer therapy, and vaccine development. However, replication-competent adenovirus (RCA) generated when adenoviral vectors are prepared in HEK293 cells has remained a concern. In this study, the human adenovirus-5 was modified to shorten the length of homologous sequence between the adenovirus and HEK293 genomic DNA, thereby reducing the production of RCA. The recombinant hAd5 was amplified and serially passaged 12 times in HEK293 cells. The amounts of RCA at passage 2, 4, 6, 8, 10, and 12 were detected by quantitative real-time PCR. The results demonstrated that the modification of adenoviral vector could effectively reduce the production of RCA during serial passages in HEK293 cells.
{"title":"Construction of recombinant adenovirus-5 vector to prevent replication-competent adenovirus occurrence","authors":"Wenbo Xie, Yifei Yuan, Bo Liu, Min Liang","doi":"10.3389/av.2023.11642","DOIUrl":"https://doi.org/10.3389/av.2023.11642","url":null,"abstract":"Human adenovirus-5 (hAd5) is an important gene delivery vector, which has been widely used in various fields of biomedicine, such as gene therapy, cancer therapy, and vaccine development. However, replication-competent adenovirus (RCA) generated when adenoviral vectors are prepared in HEK293 cells has remained a concern. In this study, the human adenovirus-5 was modified to shorten the length of homologous sequence between the adenovirus and HEK293 genomic DNA, thereby reducing the production of RCA. The recombinant hAd5 was amplified and serially passaged 12 times in HEK293 cells. The amounts of RCA at passage 2, 4, 6, 8, 10, and 12 were detected by quantitative real-time PCR. The results demonstrated that the modification of adenoviral vector could effectively reduce the production of RCA during serial passages in HEK293 cells.","PeriodicalId":7205,"journal":{"name":"Acta virologica","volume":" 6","pages":""},"PeriodicalIF":1.7,"publicationDate":"2023-12-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138962631","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Based on the crystal structure of the 3C-like protease/Nsp5 (PDB ID 6W63), virtual hits were screened from a natural product compounds database—containing 407270 natural products—by using the high-throughput virtual screening (HTVS) module of Discovery Studio software, and then filtering by “Lipinski’s rule of five” from the top 20 virtual hits. Two star-hits were selected by CDOCKER results and the protein-ligand interactions with the 3CLpro were analyzed. Finally, a 100 ns molecular dynamics simulation was carried out to verify the stability of the receptor-ligand complexes. We screened potent broad-spectrum non-covalent inhibitors that could bind to the SARS-CoV-2 3CLpro active binding site from the natural product compounds library through HTVS and molecular dynamics simulations methods. The LibDock scores and -CDOCKER energy value of the star-hits were higher than the original ligands (X77) bound to 3CLpro. CNP0348829 and CNP0474002, as star-hits, can bind stably to the active site of 3CLpro, which are promising candidate compounds for the treatment of SARS-CoV-2 and provide a theoretical basis for the development of antiviral drugs. The results of the present study may be useful in the prevention and therapeutic perspectives of COVID-19. However, further in vitro and in vivo validation tests are required in the future.
{"title":"Virtual screening and molecular dynamics simulation to identify potential SARS-CoV-2 3CLpro inhibitors from a natural product compounds library","authors":"Chunchun Gan, Xiaopu Jia, Shuai Fan, Shuqing Wang, Weikai Jing, Xiaopeng Wei","doi":"10.3389/av.2023.12464","DOIUrl":"https://doi.org/10.3389/av.2023.12464","url":null,"abstract":"Based on the crystal structure of the 3C-like protease/Nsp5 (PDB ID 6W63), virtual hits were screened from a natural product compounds database—containing 407270 natural products—by using the high-throughput virtual screening (HTVS) module of Discovery Studio software, and then filtering by “Lipinski’s rule of five” from the top 20 virtual hits. Two star-hits were selected by CDOCKER results and the protein-ligand interactions with the 3CLpro were analyzed. Finally, a 100 ns molecular dynamics simulation was carried out to verify the stability of the receptor-ligand complexes. We screened potent broad-spectrum non-covalent inhibitors that could bind to the SARS-CoV-2 3CLpro active binding site from the natural product compounds library through HTVS and molecular dynamics simulations methods. The LibDock scores and -CDOCKER energy value of the star-hits were higher than the original ligands (X77) bound to 3CLpro. CNP0348829 and CNP0474002, as star-hits, can bind stably to the active site of 3CLpro, which are promising candidate compounds for the treatment of SARS-CoV-2 and provide a theoretical basis for the development of antiviral drugs. The results of the present study may be useful in the prevention and therapeutic perspectives of COVID-19. However, further in vitro and in vivo validation tests are required in the future.","PeriodicalId":7205,"journal":{"name":"Acta virologica","volume":"12 5","pages":""},"PeriodicalIF":1.7,"publicationDate":"2023-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138999298","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
TRK-fused gene (TFG, tropomyosin-receptor kinase fused gene) is known to negatively regulate the retinoic acid inducible gene (RIG)-I-like receptor (RLR)-mediated interferon (IFN)-I pathway in human cells, thereby participating in the paramyxovirus infection process. We showed that pigeon paramyxovirus type 1 (PPMV-1) infection significantly upregulates TFG expression in infected cells at an early stage. We speculated that PPMV-1 would inhibit IFN activation by upregulating a negative regulator of the IFN pathway. This hypothesis was proved when TFG protein expression was knocked down by RNAi and the replication level of PPMV-1 virus decreased, which indicated that TFG upregulation in the early infection stage benefit virus replication. We next used the IFN-β promoter reporter system to evaluate the role of the TFG in the IFN pathway. The results showed that the TFG inhibited the IFN-β expression stimulated by RIG-I, MAVS (mitochondrial antiviral signaling protein) and TANK-binding kinase 1 (TBK1), but did not inhibit IFN-β activated by the interferon regulatory transcription factor 3 (IRF3), indicating that TFG may affect the function of TBK1, which play an important role in phosphorylation of the IRF3. Further experiments showed that the TFG inhibited the phosphorylation of TBK1, resulting in IRF3 being unable to be phosphorylated. Subsequent experiments on IFN pathway activation confirmed that the IRF3 phosphorylation level was significantly downregulated after overexpression of TFG, while the IFN-β promoter reporting experiment showed that TFG did not directly inhibit the IFN response activated by IRF3. This confirmed that TFG protein negatively regulates the IFN-β pathway by inhibiting TBK1 phosphorylation.
{"title":"The TRK-fused gene negatively regulates interferon signaling by inhibiting TBK1 phosphorylation during PPMV-1 infection","authors":"Ye Tian, Ruixue Xue, Cuilian Yu, Liping Liu, Shumin Chen, Junfeng Lv","doi":"10.3389/av.2023.11607","DOIUrl":"https://doi.org/10.3389/av.2023.11607","url":null,"abstract":"TRK-fused gene (TFG, tropomyosin-receptor kinase fused gene) is known to negatively regulate the retinoic acid inducible gene (RIG)-I-like receptor (RLR)-mediated interferon (IFN)-I pathway in human cells, thereby participating in the paramyxovirus infection process. We showed that pigeon paramyxovirus type 1 (PPMV-1) infection significantly upregulates TFG expression in infected cells at an early stage. We speculated that PPMV-1 would inhibit IFN activation by upregulating a negative regulator of the IFN pathway. This hypothesis was proved when TFG protein expression was knocked down by RNAi and the replication level of PPMV-1 virus decreased, which indicated that TFG upregulation in the early infection stage benefit virus replication. We next used the IFN-β promoter reporter system to evaluate the role of the TFG in the IFN pathway. The results showed that the TFG inhibited the IFN-β expression stimulated by RIG-I, MAVS (mitochondrial antiviral signaling protein) and TANK-binding kinase 1 (TBK1), but did not inhibit IFN-β activated by the interferon regulatory transcription factor 3 (IRF3), indicating that TFG may affect the function of TBK1, which play an important role in phosphorylation of the IRF3. Further experiments showed that the TFG inhibited the phosphorylation of TBK1, resulting in IRF3 being unable to be phosphorylated. Subsequent experiments on IFN pathway activation confirmed that the IRF3 phosphorylation level was significantly downregulated after overexpression of TFG, while the IFN-β promoter reporting experiment showed that TFG did not directly inhibit the IFN response activated by IRF3. This confirmed that TFG protein negatively regulates the IFN-β pathway by inhibiting TBK1 phosphorylation.","PeriodicalId":7205,"journal":{"name":"Acta virologica","volume":"97 7","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-11-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135869156","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Despite the urgent need for effective antivirals against SARS-CoV-2 to mitigate the catastrophic impact of the COVID-19 pandemic, favipiravir and ivermectin are among the common repurposed drugs that have been provisionally used in some countries. There have been clinical trials with mixed results, and therefore, it is still inconclusive whether they are effective or should be dismissed. It is plausible that the lack of clear-cut clinical benefits was due to the finding of only marginal levels of in vivo antiviral activity. An obvious way to improve the activity of antivirals is to use them in synergistic combinations. The in vitro antiviral activity of the combinations of favipiravir, ivermectin, niclosamide, and chloroquine against SARS-CoV-2 was assessed in Vero E6 cells and the lung epithelial cell, Calu-3. Here we show that favipiravir and ivermectin had synergistic effects against SARS-CoV-2 in Vero E6 cells. In addition, we found that favipiravir had an additive effect with niclosamide, another repurposed anti-parasitic drug with anti-SARS-CoV-2 activity. However, the anti-SARS-CoV-2 activity of favipiravir was drastically reduced when evaluated in Calu-3 cells. This suggested that this cell type might not be able to metabolize favipiravir into its active form and that this deficiency in some cell types may affect the in vivo efficacy of this drug. Favipiravir and ivermectin show the best synergistic effect. This combination is being tested in a randomized controlled clinical trial (NCT05155527).
{"title":"Favipiravir and ivermectin show in vitro synergistic antiviral activity against SARS-CoV-2","authors":"Kunlakanya Jitobaom, Chompunuch Boonarkart, Suwimon Manopwisedjaroen, Nuntaya Punyadee, Suparerk Borwornpinyo, Arunee Thitithanyanont, Panisadee Avirutnan, Prasert Auewarakul","doi":"10.3389/av.2023.12265","DOIUrl":"https://doi.org/10.3389/av.2023.12265","url":null,"abstract":"Despite the urgent need for effective antivirals against SARS-CoV-2 to mitigate the catastrophic impact of the COVID-19 pandemic, favipiravir and ivermectin are among the common repurposed drugs that have been provisionally used in some countries. There have been clinical trials with mixed results, and therefore, it is still inconclusive whether they are effective or should be dismissed. It is plausible that the lack of clear-cut clinical benefits was due to the finding of only marginal levels of in vivo antiviral activity. An obvious way to improve the activity of antivirals is to use them in synergistic combinations. The in vitro antiviral activity of the combinations of favipiravir, ivermectin, niclosamide, and chloroquine against SARS-CoV-2 was assessed in Vero E6 cells and the lung epithelial cell, Calu-3. Here we show that favipiravir and ivermectin had synergistic effects against SARS-CoV-2 in Vero E6 cells. In addition, we found that favipiravir had an additive effect with niclosamide, another repurposed anti-parasitic drug with anti-SARS-CoV-2 activity. However, the anti-SARS-CoV-2 activity of favipiravir was drastically reduced when evaluated in Calu-3 cells. This suggested that this cell type might not be able to metabolize favipiravir into its active form and that this deficiency in some cell types may affect the in vivo efficacy of this drug. Favipiravir and ivermectin show the best synergistic effect. This combination is being tested in a randomized controlled clinical trial (NCT05155527).","PeriodicalId":7205,"journal":{"name":"Acta virologica","volume":"29 6","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-11-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135934155","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Dongjin Choi, Megha Rai, Amit Rai, Mami Yamazaki, Yoonsoo Hahn
The genus Potyvirus (the family Potyviridae ) is the largest group of plant-infecting viruses transmitted by aphids. Through high-throughput RNA sequencing analysis of asymptomatic samples of Aconitum carmichaelii , a significant medicinal herb in Asia, we identified the genome sequences of two RNA viruses, tentatively named Aconitum potyvirus 1 (AcoPV1) and Aconitum potyvirus 2 (AcoPV2). The genomes of AcoPV1 and AcoPV2 encode polyproteins composed of 3,069 and 3,054 amino acids, respectively. Sequence comparisons and phylogenetic analyses established that AcoPV1 and AcoPV2 represent unique, novel members within the genus Potyvirus . The estimated RNA polymerase slippage rates at the GAAAAAA motif, responsible for the production of P3N-PIPO or P3N-ALT trans-frame fusion proteins, were 0.79% in AcoPV1 and 1.38% in AcoPV2. The RNA reads of AcoPV1 and AcoPV2 were predominantly found in the leaf and flower tissues, indicating potential feeding preferences of vectors for these viruses. These findings demonstrate the effectiveness of high-throughput RNA sequencing in not only uncovering novel potyviruses, but also in elucidating their genomic dynamics within host plants.
{"title":"Discovery of two novel potyvirus genome sequences by high-throughput RNA sequencing in Aconitum carmichaelii tissue samples","authors":"Dongjin Choi, Megha Rai, Amit Rai, Mami Yamazaki, Yoonsoo Hahn","doi":"10.3389/av.2023.11782","DOIUrl":"https://doi.org/10.3389/av.2023.11782","url":null,"abstract":"The genus Potyvirus (the family Potyviridae ) is the largest group of plant-infecting viruses transmitted by aphids. Through high-throughput RNA sequencing analysis of asymptomatic samples of Aconitum carmichaelii , a significant medicinal herb in Asia, we identified the genome sequences of two RNA viruses, tentatively named Aconitum potyvirus 1 (AcoPV1) and Aconitum potyvirus 2 (AcoPV2). The genomes of AcoPV1 and AcoPV2 encode polyproteins composed of 3,069 and 3,054 amino acids, respectively. Sequence comparisons and phylogenetic analyses established that AcoPV1 and AcoPV2 represent unique, novel members within the genus Potyvirus . The estimated RNA polymerase slippage rates at the GAAAAAA motif, responsible for the production of P3N-PIPO or P3N-ALT trans-frame fusion proteins, were 0.79% in AcoPV1 and 1.38% in AcoPV2. The RNA reads of AcoPV1 and AcoPV2 were predominantly found in the leaf and flower tissues, indicating potential feeding preferences of vectors for these viruses. These findings demonstrate the effectiveness of high-throughput RNA sequencing in not only uncovering novel potyviruses, but also in elucidating their genomic dynamics within host plants.","PeriodicalId":7205,"journal":{"name":"Acta virologica","volume":"4 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135968977","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Jun Zhang, Zeming Guo, Yulin Zhao, Yida Yang, Pan Huang, Ning Wang, Zhuoyu Qian, Menghan He, Jianmin Wu, Lv Luo, Zhongsheng Li, Chungen Pan
Since the last century, the spread of the genotype 2 classical swine fever virus (CSFV) has caused significant issues for the pig breeding industries. Ideal strategies for controlling CSFV include vaccination and keeping farms free of CSFV. For vaccination, several attenuated CSFV viruses originating from genotype 1 are widely used; for the latter, accurate diagnosis is required for detection of the CSFV infection. Nucleic acid testing for CSFV usually uses tonsil samples, which requires an inconvenient sampling operation that injures pigs. Commercial serological tests for CSFV antibodies or antigens are unable to distinguish the genotype for originating virus. In this study, 20 mAbs were developed from the mice hybridoma cells. Four of the mAbs were identified to have the ability to only recognize the peptides derived from sub-genotype 2.1 strain, and two of them, MM1 and MM5, were further studied to identify critical binding sites (epitopes) on the E2 protein of CSFV. A total of 353 genotype 2 collections were made worldwide in GeneBank, 90.9% of which contained MM1 or MM5 epitopes. Moreover, 95.1% of sub-genotype 2.1 isolations contained MM5 epitope. Therefore, MM1 and MM5 have the potential to be developed as a diagnostic tool for detection of genotype 2 virus antigen by indirect ELISA or antibodies by competitive ELISA.
{"title":"Identification of novel monoclonal antibodies specific for the conserved epitopes in the E2 protein of genotype 2 classical swine fever virus: implication for differential diagnosis","authors":"Jun Zhang, Zeming Guo, Yulin Zhao, Yida Yang, Pan Huang, Ning Wang, Zhuoyu Qian, Menghan He, Jianmin Wu, Lv Luo, Zhongsheng Li, Chungen Pan","doi":"10.3389/av.2023.12124","DOIUrl":"https://doi.org/10.3389/av.2023.12124","url":null,"abstract":"Since the last century, the spread of the genotype 2 classical swine fever virus (CSFV) has caused significant issues for the pig breeding industries. Ideal strategies for controlling CSFV include vaccination and keeping farms free of CSFV. For vaccination, several attenuated CSFV viruses originating from genotype 1 are widely used; for the latter, accurate diagnosis is required for detection of the CSFV infection. Nucleic acid testing for CSFV usually uses tonsil samples, which requires an inconvenient sampling operation that injures pigs. Commercial serological tests for CSFV antibodies or antigens are unable to distinguish the genotype for originating virus. In this study, 20 mAbs were developed from the mice hybridoma cells. Four of the mAbs were identified to have the ability to only recognize the peptides derived from sub-genotype 2.1 strain, and two of them, MM1 and MM5, were further studied to identify critical binding sites (epitopes) on the E2 protein of CSFV. A total of 353 genotype 2 collections were made worldwide in GeneBank, 90.9% of which contained MM1 or MM5 epitopes. Moreover, 95.1% of sub-genotype 2.1 isolations contained MM5 epitope. Therefore, MM1 and MM5 have the potential to be developed as a diagnostic tool for detection of genotype 2 virus antigen by indirect ELISA or antibodies by competitive ELISA.","PeriodicalId":7205,"journal":{"name":"Acta virologica","volume":"198 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136209270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Katarína Čurová, Viera Lovayová, Mária Nagyová, Leonard Siegfried, Viliam Donič, Gert de Vos
The reverse transcription polymerase chain reaction (RT-PCR) is considered the gold standard method for the detection of viruses in a clinic. The aim of this study was to compare the ability of conventional RT-PCR test (FTD TM SARS-CoV-2 Test) and laboratory-developed ultra-fast PCR test (NextGenPCR TM SARS-CoV-2 RT-PCR Reagent Kit) to detect the coronavirus SARS-CoV-2 causing COVID-19. A total of 318 nasopharyngeal swab specimens were collected from people under investigation for COVID-19. Despite the collection of two swab specimens from each patient and their different processing, the analysis showed an overall agreement of 95.9% between the conventional and laboratory-developed tests. The positive percentage agreement was 90.5% (114/126) and the negative percentage agreement was 99.5% (191/192). The ultra-fast NextGenPCR method does not require the isolation of RNA, provides a result of 20–96 specimens within 57–82 min after sampling, and offers a simple procedure of sample processing, analysis, and evaluation. Our results indicate that this method can be considered a potential diagnostic method for the detection of SARS-CoV-2 virus in hospitals, healthcare facilities, and research laboratories.
{"title":"Detection of SARS-CoV-2 using a laboratory-developed ultra-fast NextGenPCR test versus a conventional RT-PCR test","authors":"Katarína Čurová, Viera Lovayová, Mária Nagyová, Leonard Siegfried, Viliam Donič, Gert de Vos","doi":"10.3389/av.2023.11588","DOIUrl":"https://doi.org/10.3389/av.2023.11588","url":null,"abstract":"The reverse transcription polymerase chain reaction (RT-PCR) is considered the gold standard method for the detection of viruses in a clinic. The aim of this study was to compare the ability of conventional RT-PCR test (FTD TM SARS-CoV-2 Test) and laboratory-developed ultra-fast PCR test (NextGenPCR TM SARS-CoV-2 RT-PCR Reagent Kit) to detect the coronavirus SARS-CoV-2 causing COVID-19. A total of 318 nasopharyngeal swab specimens were collected from people under investigation for COVID-19. Despite the collection of two swab specimens from each patient and their different processing, the analysis showed an overall agreement of 95.9% between the conventional and laboratory-developed tests. The positive percentage agreement was 90.5% (114/126) and the negative percentage agreement was 99.5% (191/192). The ultra-fast NextGenPCR method does not require the isolation of RNA, provides a result of 20–96 specimens within 57–82 min after sampling, and offers a simple procedure of sample processing, analysis, and evaluation. Our results indicate that this method can be considered a potential diagnostic method for the detection of SARS-CoV-2 virus in hospitals, healthcare facilities, and research laboratories.","PeriodicalId":7205,"journal":{"name":"Acta virologica","volume":"45 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136293713","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The biological consequences of viral infection result from biochemical, physiological, structural, morphological and genetic changes in infected cells. In productive infections, virus-induced biological changes in cells may be closely related to the efficiency of viral replication or to the recognition of these cells by the immune system. These changes are usually associated with cytocidal viruses, as in the case of the pandemic coronavirus SARS-CoV-2, which causes COVID-19. Many of these changes are required for effective viral replication. The physiological state of living cells has a significant impact on the outcome of viral infection, as the host cell provides the synthetic machinery, key regulatory molecules and precursors for newly synthesised viral proteins and nucleic acids. This review focuses on novel target cell types for SARS-CoV-2 exposure outside the respiratory tract. Findings and examples are collected that provide information on virus-cell interactions. The identification of unusual target cells for SARS-CoV-2 may help to explain the diverse symptoms in COVID-19 patients and the long-lasting effects after infection. In particular, the discovery of previously undescribed target cells for SARS-CoV-2 action needs to be considered to improve treatment of patients and prevention of infection.
{"title":"The effects of SARS-CoV-2 on susceptible human cells","authors":"Zinaida Klestova","doi":"10.3389/av.2023.11997","DOIUrl":"https://doi.org/10.3389/av.2023.11997","url":null,"abstract":"The biological consequences of viral infection result from biochemical, physiological, structural, morphological and genetic changes in infected cells. In productive infections, virus-induced biological changes in cells may be closely related to the efficiency of viral replication or to the recognition of these cells by the immune system. These changes are usually associated with cytocidal viruses, as in the case of the pandemic coronavirus SARS-CoV-2, which causes COVID-19. Many of these changes are required for effective viral replication. The physiological state of living cells has a significant impact on the outcome of viral infection, as the host cell provides the synthetic machinery, key regulatory molecules and precursors for newly synthesised viral proteins and nucleic acids. This review focuses on novel target cell types for SARS-CoV-2 exposure outside the respiratory tract. Findings and examples are collected that provide information on virus-cell interactions. The identification of unusual target cells for SARS-CoV-2 may help to explain the diverse symptoms in COVID-19 patients and the long-lasting effects after infection. In particular, the discovery of previously undescribed target cells for SARS-CoV-2 action needs to be considered to improve treatment of patients and prevention of infection.","PeriodicalId":7205,"journal":{"name":"Acta virologica","volume":"23 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135386722","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Serological testing is a powerful tool for analyzing the infectious disease burden landscape. Therefore, this study aimed to determine the seroprevalence against SARS-CoV-2 in the population of the municipality of Kragujevac, Serbia, with a particular reference to silent infections. A total of 4,804 participants over 19 years of age were randomly sampled for population-based seroprevalence research. Anti-N IgG antibodies were measured using rapid serological tests (UNscience ® ). The population was divided into four Cohorts, according to the history of SARS-CoV-2 infection and vaccination status with the whole inactivated virus vaccine BBIBP-CorV (Vero Cell ® , Sinopharm), as follows: Cohort I—confirmed SARS-CoV-2 infection, not vaccinated with the BBIBP-CorV vaccine; Cohort II—without confirmed SARS- CoV-2 infection, vaccinated with the BBIBP-CorV vaccine; Cohort III—confirmed SARS-CoV-2 infection, vaccinated with the BBIBP-CorV vaccine; Cohort IV—without confirmed SARS-CoV-2 infection, not vaccinated with the BBIBP-CorV vaccine (silent immunization). Cohorts I and IV included patients vaccinated with vaccines other than the BBIBP-CorV vaccine. The results showed that the overall prevalence of anti-N IgG antibodies was 56.5%, with the highest seroprevalence in Cohort III at 85.8%. In Cohort IV, the prevalence of anti-N IgG antibodies was 40.7%, attributed to silent immunization. The results also suggest that the prevalence of anti-N IgG antibodies decreased over time but remained detectable for more than 12 months in Cohort I. Since currently, there is no data on silent infection frequency in our country, these findings may provide insight into the extent of silent infections in the Serbian population.
{"title":"Silent SARS-CoV-2 infection: seroprevalence study of SARS-CoV-2 anti- nucleocapsid IgG antibodies in Kragujevac, Serbia","authors":"Neda Cicaric, Vanja Canovic, Milica Stojkovic, Sanja Matic, Srdjan Stefanovic, Suzana Popovic, Danijela Todorovic, Natasa Djordjevic, Biljana Radenkovic, Marko Radenkovic, Vasilije Antic, Dejan Baskic","doi":"10.3389/av.2023.11996","DOIUrl":"https://doi.org/10.3389/av.2023.11996","url":null,"abstract":"Serological testing is a powerful tool for analyzing the infectious disease burden landscape. Therefore, this study aimed to determine the seroprevalence against SARS-CoV-2 in the population of the municipality of Kragujevac, Serbia, with a particular reference to silent infections. A total of 4,804 participants over 19 years of age were randomly sampled for population-based seroprevalence research. Anti-N IgG antibodies were measured using rapid serological tests (UNscience ® ). The population was divided into four Cohorts, according to the history of SARS-CoV-2 infection and vaccination status with the whole inactivated virus vaccine BBIBP-CorV (Vero Cell ® , Sinopharm), as follows: Cohort I—confirmed SARS-CoV-2 infection, not vaccinated with the BBIBP-CorV vaccine; Cohort II—without confirmed SARS- CoV-2 infection, vaccinated with the BBIBP-CorV vaccine; Cohort III—confirmed SARS-CoV-2 infection, vaccinated with the BBIBP-CorV vaccine; Cohort IV—without confirmed SARS-CoV-2 infection, not vaccinated with the BBIBP-CorV vaccine (silent immunization). Cohorts I and IV included patients vaccinated with vaccines other than the BBIBP-CorV vaccine. The results showed that the overall prevalence of anti-N IgG antibodies was 56.5%, with the highest seroprevalence in Cohort III at 85.8%. In Cohort IV, the prevalence of anti-N IgG antibodies was 40.7%, attributed to silent immunization. The results also suggest that the prevalence of anti-N IgG antibodies decreased over time but remained detectable for more than 12 months in Cohort I. Since currently, there is no data on silent infection frequency in our country, these findings may provide insight into the extent of silent infections in the Serbian population.","PeriodicalId":7205,"journal":{"name":"Acta virologica","volume":"69 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134887163","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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