Abstract The emergence and circulation of duck reovirus have caused severe threats to domestic waterfowl production because of the lethal infections they cause in ducks and geese. However, the evolution of circulating duck reoviruses and their replication and pathogenicity in domestic birds have not been fully investigated. In this study, we identified and isolated six duck reoviruses from clinical samples of sick or deceased farmed ducks and geese and sequenced their full genomes. Phylogenetic analysis revealed the evolutionary landscape of duck reoviruses and the complex reassortment of these circulating viruses with avian orthoreovirus and Muscovy duck reovirus. Animal infection studies revealed differences in the replication and pathogenicity of the reoviruses identified in this study in ducks, geese and chickens. Lethal infection with highly pathogenic viruses causes severe focal necrosis and hemorrhage in the liver, spleen, bursa of Fabricius and thymus, resulting in high mortality in inoculated birds. Importantly, chickens are susceptible to circulating duck reovirus, highlighting the potential risk of duck reovirus infection in chickens. Our study revealed the evolution, pathogenicity and potential cross-species transmission risk of duck reoviruses, further emphasizing the importance of continued and systemic surveillance at the interface of domestic waterfowl and chickens.
Multiple studies on PIK3CA mutations in breast cancer (BC) had been performed, which showed the controversial results among different countries and races even those from the same country. The present study aimed to explore the PIK3CA gene mutation status in BC patients in Northwest China and reveal the relationship between PIK3CA mutations and clinicopathological features along with prognosis.1002 BC patients from Northwest China were recruited in this study, genomic DNA was extracted from formalin-fixed paraffin-embedded (FFPE) tumor tissues, and hotspot mutations in the exon 9 and 20 of PIK3CA gene were detected by ARMS-PCR.PIK3CA mutations were found in 31.2% (313/1002) of BC patients, among them 66.1% were mutations in exon 20% and 32.6% were mutations in exon 9. H1047R was the most common mutation type, accounting for 56.5% of the total mutated samples. Significant correlations were observed between PIK3CA mutation status and age (P = 0.035), histopathologic types (P = 0.004), pathological grade (P = 0.013), ER positivity (P < 0.001), PR positivity (P < 0.001), molecular subtypes (P = 0.004) and family history (P = 0.007). Cox multivariate analysis showed that patients with mutations in exon 9 or 20 had shorter DFS and OS than wild-type patients. Those with exon 9 mutations subgroup had the worst prognosis. Interestingly, patients with H1047L mutation had the best prognosis than others.PIK3CA mutations could be used as an indicator of clinical outcome or targeted therapy for multiple breast cancer subgroups in Northwest China.
Porcine deltacoronavirus (PDCoV) has emerged and spread throughout the porcine industry in many countries over the last 6 years. PDCoV caused watery diarrhoea, vomiting and dehydration in newborn piglets. A sensitive diagnostic method would be beneficial to the prevention and control of PDCoV infection. Recombinase polymerase amplification (RPA) is an isothermal amplification method which has been widely used for virus detection. A probe-based reverse transcription RPA (RT-RPA) assay was developed for real-time detection of PDCoV. The amplification can be finished in 20 min and fluorescence monitoring was performed by a portable device. The lowest detection limit of the PDCoV RT-RPA assay was 100 copies of RNA molecules per reaction; moreover, the RT-RPA assay had no cross-reaction with other common swine viruses. The clinical performance of the RT-RPA assay was evaluated using 108 clinical samples (54 intestine specimens and 54 faecal swab specimens). The coincidence rate of the detection results for clinical samples between RT-RPA and RT-qPCR was 97.2%. In summary, the real-time RT-RPA assay offers a promising alternative to RT-qPCR for point-of-care detection of PDCoV.
Using Luminex xMAP (x = analyte, MAP = multi-analyte profiling) technology, a serological method for the simultaneous detection of antibodies to Newcastle disease virus (NDV) and avian influenza virus (AIV) was established. Nano-magnetic beads coated with purified NDV protein and AIV nucleoprotein were incubated with serum samples. Using biotinylated rabbit anti-chicken IgY and streptavidin-R-phycoerythrin, the optical signals measured by a Luminex 200 detection system indicated the quantification of NDV or AIV antibodies in the serum. Specific pathogen-free (SPF) chicken serum was used as a negative control. The Luminex xMAP assay developed in this study demonstrated high specificity as there was no cross-reaction with antibodies to infectious laryngotracheitis virus, infectious bronchitis virus, infectious bursal disease virus, avian leukosis virus, and Marek's disease virus. The results from reproducibility experiments showed that intra-coefficients of variation were 3.36 and 9.23% and inter-coefficients of variation were 6.50 and 7.66% for NDV and AIV, respectively. The results also indicated that the Luminex xMAP assay was 16 times more sensitive for NDV antibody detection and 1,024 times more sensitive for AIV antibody detection compared to the enzyme-linked immunosorbent assay (ELISA). A total of 300 chicken serum samples were subjected to both Luminex xMAP assay and ELISA, showing the coincidence rates of 98.67 and 98% for NDV and AIV antibody detection, respectively. This study provides a new method for the simultaneous detection NDV and AIV antibodies in the serum with high specificity and sensitivity.
Abstract Background: Ectromelia virus (ECTV) is the pathogen of mousepox, which has a limited host range and a high mortality rate. In the past century, ECTV has affected laboratory mouse colonies worldwide. Recombinant polymerase amplification (RPA), which is widely used in virus detection, is an isothermal amplification method. Results: In this study, a probe-based RPA detection method was established for rapid and sensitive detection of ECTV.Primers were designed for the highly conserved region of crmD gene, the main core protein of recessive poxvirus, and standard plasmids were constructed.The sensitivity of the RPA assay is 100 copies of the genome per reaction. In addition, the method showed high specificity and did not cross-react with other common mouse viruses.Therefore, the practicability of the RPA method in the field was confirmed by the detection of 52 clinical samples. The real-time RPA assay was very similar to the ECTV real-time PCR assay, with 100% agreement. Conclusions: In conclusion, this RPA assay, especially in low-resource settings, provides a novel alternative for sensitive, simple, and specific identification of ECTV.
Abstract Background: Budgerigar fledgling disease virus (BFDV) poses a serious threat to the Chinese psittacine industry, causing enormous economic losses. This study aims to reveal the etiological role of BFDV and evaluate the molecular characterization. Results: We report on BFDV, designated SC-YB19, which had an 18-nucleotide (nt) deletion in the enhancer region, corresponding to the sequence position 164–181 nt, when compared with other BFDV strains. Sequence analyses suggested that 19 nucleotide substitutions were identified with the domestic strains, APV7 and AF118150. Phylogenetic analysis indicated that SC-YB19, along with three domestic strains, formed a unique cluster, and were closely related to Polish isolates. Conclusion: Taken together, these results demonstrate that a BFDV genotype variation was co-circulating in China, and provide important insights on evolution of BFDV.
The porcine epidemic diarrhea virus (PEDV) has caused severe economic losses in the pig industry. Since its discovery, the virus has spread in pig herds for more than 50 years. Many new features have been found in the PEDV spike genes. In this study, 123 representative S genes were used to analyze their characteristics across strains. Phylogenetic analysis revealed that PEDV can be divided into nine groups: G1a, G1b, G1c, G1d, G2a, G2b, G2c, G2d, and G3. In addition, 21 different lengths of the S gene were found. Analysis of the amino acid insertion and deletion sites revealed that most deletions and insertions occurred in the loops of the spike quaternary structure, primarily in the D0 and N‐terminal domains (NTDs). According to the above results, PEDV has undergone considerable evolution, possibly under the immune pressure of vaccination. These results are highly important for understanding the current epidemic situation of PEDV.
Seneca Valley virus (SVV) is an oncolytic virus, which belongs to the Picornaviridae family, that causes blisters on the nose and hooves, affecting the production performance of pigs. However, the function of proinflammatory cytokines and chemokines in SVV infection is still unclear. In our study, SVV infection could induce a high expression of proinflammatory cytokines interleukin (IL)-1α, IL-1β, and tumor necrosis factor α (TNF-α) and chemokines, including chemokine (C-C motif) ligand 2 (CCL2), chemokine (C-C motif) ligand 5 (CCL5), and chemokine (C-X-C motif) ligand 10 (CXCL10). Interfered genes of IL-1α, IL-1β, and TNF-α inhibited virus replication, but interfered genes of CCL2, CCL5, and CXCL10 promoted virus replication. These results indicate that proinflammatory cytokines and chemokines are involved in SVV infection; this will be beneficial to explore the pathogenesis and cytokine therapy of SVV.Le virus de la Vallée de Seneca (SVV) est un virus oncolytique, qui appartient à la famille des Picornaviridae, qui provoque des cloques sur le nez et les sabots, affectant les performances de production des porcs. Cependant, la fonction des cytokines pro-inflammatoires et des chimiokines dans l’infection par le SVV n’est toujours pas claire. Dans notre étude, l’infection par le SVV pourrait induire une forte expression des cytokines pro-inflammatoires interleukine (IL)-1α, IL-1β, et du facteur de nécrose tumorale α (TNF-α) et des chimiokines, y compris la chimiokine (motif C-C) ligand 2 (CCL2), chimiokine (motif C-C) ligand 5 (CCL5) et chimiokine (motif C-X-C) ligand 10 (CXCL10). Les gènes interférés d’IL-1α, IL-1β et TNF-α inhibent la réplication virale, mais les gènes interférents de CCL2, CCL5 et CXCL10 favorisent la réplication virale. Ces résultats indiquent que les cytokines pro-inflammatoires et les chimiokines sont impliquées dans l’infection par le SVV; cela sera bénéfique pour explorer la pathogenèse et la thérapie par cytokines du SVV.(Traduit par Docteur Serge Messier).
Abstract Background Ectromelia virus (ECTV) is the causative agent of mousepox in mice. In the past century, ECTV was a serious threat to laboratory mouse colonies worldwide. Recombinase polymerase amplification (RPA), which is widely used in virus detection, is an isothermal amplification method. Results In this study, a probe-based RPA detection method was established for rapid and sensitive detection of ECTV.Primers were designed for the highly conserved region of the crmD gene, the main core protein of recessive poxvirus, and standard plasmids were constructed. The lowest detection limit of the ECTV RT- RPA assay was 100 copies of DNA mol-ecules per reaction. In addition, the method showed high specificity and did not cross-react with other common mouse viruses.Therefore, the practicability of the RPA method in the field was confirmed by the detection of 135 clinical samples. The real-time RPA assay was very similar to the ECTV real-time PCR assay, with 100% agreement. Conclusions In conclusion, this RPA assay offers a novel alternative for the simple, sensitive, and specific identification of ECTV, especially in low-resource settings.
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