Bing du xue bao = Chinese journal of virology最新文献

microRNAs (miRNAs) not only play the key roles in regulation of the growth and development of mosquito, but also has an important function in interaction between the pathogen and vector. So, miRNAs can be used as the molecular target for development of an alternative method for mosquito and mosquito-borne disease control. However, an effective delivery system is still need. Mosquito densovirus have the potential for vector control as transducing agents to express foreign toxins or small interfering RNAs molecules in vitro and in vivo. In this study, we report the development of a recombinant Aedes albopictus densovirus-3(AalDV-3) miRNA sponge expression system, using an intronic miRNA sponge expression strategy. To test the inhibition effect of recombinant virus medicated miRNA sponge on endogenous miR-210, Aedes aegypti Aag2 cell and 1st-2nd larvae were infected, respectively. The splicing of the intronic miRNA sponge expression constructs in vitro and in vivo were tested by RT-PCR with intron span primers, and the relatively expression level of miR-210 was confirmed by qPCR. As results,AalDV-3can be used to decrease endogenous miRNAs by generating an antisense sponge in vitro and in vivo, which represents a tool for the functional analysis of mosquito genes and lays the foundation for the application of densovirus for vector control.

To explore the effect of porcine epidemic diarrhea virus(PEDV)on microRNA expression profiles of porcine kidney 15cell(PK-15),total RNA was isolated from PK-15 cells with or without PEDV infection. Then, we obtained the miRNAs by using solexa sequencing technology and analyzed these differentially expressed miRNAs. Heatmap cluster analysis and GO (ontology, GO) (Gene function, MF, Molecular) analysis was performed on the significant differences in expression of the miRNA, and 10 significant differences in expression of miRNA were selected by RT-qPCR. The result showed 214 kinds of microRNAs (miRNAs) expression levels were significantly different in PEDV-infected cells, compared with normal PK-15 cells. Among of them,175 kinds of miRNAs were significantly up-regulated while 39 kinds of miRNAs were significantly down-regulated. Furthermore, qPCR results of the expression trends of miRNA were similar with that of solexa sequencing. Heatmap cluster analysis showed that the vast majority of the viral groups were differentially expressed in miRNA compared with the control group, Go analysis showed that miRNAs are widely involved in combination, binding protein, protein kinase activity, transfer enzyme activity, phosphorus containing radicals transfer, phosphotransferase enzyme activity and other biological effects. The expression trends of RT-qPCR verified by miRNA were consistent with the high throughput sequencing results. The results showed that the swine epidemic diarrhea virus infection had a significant impact on the level of miRNA expression in PK-15 cells, thus providing a new idea for the further study of the miRNA preparation for the treatment of PEDV.

The first reported group A rotavirus(RVA)G9 strain T203 in mainland China detected from an infant in Beijing in 1994 clustered within VP7 lineage Ⅵ(G9-Ⅵ),whereas the common RVA G9 belong to VP7 lineage Ⅲ(G9-Ⅲ) worldwide. Interestingly, since it was first reported in 1994 there was no G9-Ⅵ circulating in Beijing, until an unexpected G9-Ⅵ strain was identified by sequencing in 2010 RVA surveillance. This present study was to develop a convenient and effective dot-blot hybridization method for differentiating G9-Ⅲ and G9-Ⅵ rotaviruses, to investigate the re-emerging circulating distribution of G9-Ⅵ RVAs in outpatient children with diarrhea from 2011 to 2012in Beijing. By multiple-sequence aligning and analyzing the collected VP7 gene nucleotide sequences of G9 RVAs worldwide from GenBank database using Clustal W software, a region within VP7 gene which is highly divergent between G9-Ⅲ and G9-Ⅵ rotaviruses and conserved within themselves was selected as probe. One pair of common primers at both side of this probe region was designed, and used for synthesizing DIG labeled G9-Ⅲ and G9-Ⅵ probes with PCR, respectively. Subsequently, RVA G and P genotypes were identified as previously described. Then G9-Ⅲ and G9-Ⅵ were further differentiated from G9 RVAs using dot-blot hybridization method established in this study. It was showed that G9 was the most prevalent genotype(43.5%),followed by G3(30.5%)、G1(12.2%)and G2(11.5%),and no G4 genotype was detected.Interestingly,G9-Ⅵ was the most predominant(96.5%) type, while only 3.5% was G9-Ⅲ among these G9 rotaviruses.Phylogenetically,G9-Ⅵ rotaviruses in this study clustered closely with human G9-Ⅵ rotaviruses which were more recently re-emerging in several countries including China around the world as well as porcine G9-Ⅵ rotavirus strain F7P4 in Canada, whereas they clustered distantly to worldwide common G9-Ⅲ strains.G9-Ⅵ rotaviruses were re-emerging in the world, whether it gained stronger spreading ability or virulence than ever common G9-Ⅲ during evolution with porcine G9-Ⅵ rotaviruses need further analysis.

Several viruses have been found in non-human primates, among which some are pathogenic to humans. To further characterize the spectrum of viruses present in wild rhesus monkeys, MiSeq high-throughput sequencing were used to analyze 280 fecal samples collected from Guangxi, China. A total of 23,372,679 reads were obtained, of which 4,641 were annotated to 27 viral families or subfamilies, including five vertebrate viruses families(78.2%),six insect virus families(5.5%),eleven plant virus families(10.4%),and other viruses(9.8%).Further analysis revealed that these reads best fit with the sapelovirus, enterovirus, parvovirus, adeno-associated virus read sequences shared a high similarity with the known viruses. However, some reads presented obvious differences from these viruses. Moreover, PCR amplification was conducted to confirm these potentially novel viruses. This study had explored the viral spectrum of rhesus monkey feces in the Guangxi area, which laid the foundation for the potential public health significance of these viruses.

We aimed to develop a novel laboratory assay for the detection of Middle East respiratory syndrome coronavirus (MERS-CoV) infection .Several novel multiplex real-time RT-PCR assays were developed using the upE,ORF1 band N2genes of MERS-CoV as targets; the novel assays were compared with previous monoplex real-time RT-PCR assays. For validation, we tested a MERS-CoV strain (hCoVEMC), clinical specimens from patients with fever in Shanghai, and specimens from the first imported MERS case in China. The detection limit of the novel multiplex real-time RT-PCR assays was 10 PFU of MERS-CoV per ml, the same as that in monoplex real-time RT-PCR assays based on upE or N2. The detection was specific for MERS-CoV. In validation using clinical samples, pharyngeal swabs from Shanghai patients were detected as negative, while swabs from the first imported MERS case in China were detected as positive. Using whole blood samples from a MERS case, better detection results were obtained with N2 as the target than upE. We conclude that all the novel assays established in this study could be used for the detection of MERS-CoV; they show potential for improvement compared with monoplex real-time RT-PCR assay based on upE.

In the current study, rabbit spleen was analysed at 12 h,24h,30 h,36h,and 48hpost-infection with hog cholera lapinized virus(C strain)using high-throughput sequencing. The rabbit genome was used as a reference to identify differentially expressed genes(DEGs)at different time points post-infection. The top 10 DEGs were filtered based on significance, and we searched for their biological functions through the Uniprot and NCBI databases. The former three time points have 10co-expressing genes, many of which have a relationship to immunity and inflammation. The latter two time points for the top 10 DEGs are identical, and B2 M,RLA-DR-ALPHA,CD74,and IGJ are involved in the antiviral immune response. GO functional annotation revealed that in biological processes at each time point,except for 24hpost-infection,the immune response has the most terms, followed by metabolism and regulation. According to the KEGG database, the DEGs for 24hpost-infection were enriched for the RIG-I-like receptor signaling pathway and the DEGs for 30hpost-infection were found to have a focal adhesion and ECM-receptor interaction pathway. Moreover, the DEGs for 36 hand 48hpost-infection have seven identical pathways, of which were directly or indirectly related to the antiviral response. These pathways included the proteasome, lysosome, ribosome, chemokine signaling pathways, B cell receptor signaling pathway, antigen processing, and presentation pathway, and the Fc gamma R-mediated phagocytosis pathway.These results provide novel insight into the gene expression in rabbit spleens post-infection with C strain, and provide a theoretical basis for further understanding of the molecular mechanisms by which rabbits adapt to infection with C strain.

HIV the pathogen responsible for the transmission of AIDS, which is associated with a high mortality rate. Vpx is expressed in HIV-2/SIV and promotes retroviral infection in specific cells. This promotion is achieved by Vpx-induced formation of the CRL4E3 complex, which removes the endogenous SAMHD1 via proteasomal degradation. Multiple domains of SAMHD1(e.g., N-terminus, nuclear localization signal, linker, HD domain, and C-terminus)are essential for Vpx-induced degradation.HIV-1that does not express Vpx is also evolved with mechanisms to bypass or suppress the antiviral function of SAMHD1,such as the tolerance against a low level of dNTPs and induction of SAMHD1 degradation through cyclin L2.Based on previous reports published chronologically, as well as the latest findings in the field, this review focuses on the mechanism of Vpx-mediated degradation of SAMHD1,and its promotion of HIV-1infection.

The Middle East respiratory syndrome coronavirus (MERS-CoV) was first isolated in 2012 from patients that died from severe pneumonia. It was another coronavirus which resulted in severe infection of humans, apart from the severe acute respiratory syndrome coronavirus. The development of an appropriate animal model is necessary to study the pathogenesis and to evaluate the intervening measures against MERS-CoV infection. To date, several small animals(e.g., mice, Syrian hamsters, and ferrets) have been used to study MERS-CoV infection. In addition, rhesus macaques, common marmosets, and dromedary camels were also utilized as the models for MERS-CoV study. The purpose of this paper is to summarize the advantages and disadvantages of the animal models mentioned above, to provide a thorough understanding of the MERS-CoV study.

Hepatitis C virus(HCV)infection is a global public health problem, primarily triggering acute and chronic liver hepatitis. Owing to the narrow host range, a suitable animal model is still lacking, hindering study of viral pathogenesis, immune control,and prophylactic vaccine development. There has no relevant evidence that homologs of HCV originating from the animal may have the potential to cross the species barrier to cause human disease until recently. Several agents discovered that new viruses related to HCV, including HCV and GBV-like viruses(belonging to the Hepacivirus and Pegivirus genera, respectively), in small wild mammals (e.g., rodents and bats)and domesticated animals(e.g., dogs, horses, and cattle). Genetic and biological characterization of these novel HCV-related viruses may provide novel insight into the origins, pathogenesis, and immune response to HCV infection in humans. In this review, we introduce the gene characteristics of HCV, concerned viruses, and many newly discovered viruses closely related to HCV-like viruses. The exploration of their natural reservoirs will be performed, and we then discuss possible theories regarding the origin of this important viral human pathogen.

This study aims to investigate the etiologic spectrum features of hand, foot and mouth disease(HFMD)and the VP1 region of coxsackievirus A6(CV-A6)in the Anyang area of China during 2013.A total of 479 HFMD fecal specimens were identified using real-time RT-PCR. The positive specimens of the different serotypes and the proportion of each serotype were calculated. A total of 10 positive of CV-A6 specimens were amplified and sequenced to obtain the profile of the VP1 region. Based on the data of the VP1 sequences from the Anyang strains and other referenced strains deposited in the NCBI database, the similarities of the nucleotide and amino-acid sequences for the VP1 region, as well as phylogenetic analyses were performed by the BIOEIDT 7.2and MEGA 5.1software packages. The results revealed that 429 positive specimens of enterovirus, including 90 positive specimens of CV-A6 were identified. The CV-A6 specimens accounted for 20.98%of the total positive specimens.CV-A6 had become the second preponderant pathogen of the HFMD in the Anyang area during 2013.It was the first record of an enterovirus serotype of which was neither CV-A16 nor EV-A71,had become one of the primary pathogens associated with HFMD in the region. A great diversity of the nucleotide and amino-acid sequences between 10 Anyang strains and a prototype strain (Gdula) was found. Compared with the Henan strain(HN421),four Anyang sequences with nucleotide and amino-acid sequences exhibited greater diversity than the other six Anyang sequences. The phylogenetic analysis showed that all of 49CV-A6 strains could be clustered into four branches. Branch D could be further clustered into two sub-branches;6 Anyang sequences belonged to sub-branch D1,and the other four belonged to sub-branch D2.