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1.
J Proteome Res ; 23(4): 1272-1284, 2024 Apr 05.
Artigo em Inglês | MEDLINE | ID: mdl-38470452

RESUMO

Gestational diabetes mellitus (GDM) with intrauterine hyperglycemia induces a series of changes in the placenta, which have adverse effects on both the mother and the fetus. The aim of this study was to investigate the changes in the placenta in GDM and its gender differences. In this study, we established an intrauterine hyperglycemia model using ICR mice. We collected placental specimens from mice before birth for histological observation, along with tandem mass tag (TMT)-labeled proteomic analysis, which was stratified by sex. When the analysis was not segregated by sex, the GDM group showed 208 upregulated and 225 downregulated proteins in the placenta, primarily within the extracellular matrix and mitochondria. Altered biological processes included cholesterol metabolism and oxidative stress responses. After stratification by sex, the male subgroup showed a heightened tendency for immune-related pathway alterations, whereas the female subgroup manifested changes in branched-chain amino acid metabolism. Our study suggests that the observed sex differences in placental protein expression may explain the differential impact of GDM on offspring.


Assuntos
Diabetes Gestacional , Hiperglicemia , Humanos , Gravidez , Feminino , Masculino , Camundongos , Animais , Placenta/metabolismo , Proteômica , Camundongos Endogâmicos ICR , Diabetes Gestacional/genética , Diabetes Gestacional/metabolismo , Hiperglicemia/genética
2.
Avian Pathol ; 52(2): 128-136, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36622371

RESUMO

Chicken parvovirus (ChPV), chicken infectious anaemia virus (CIAV) and fowl adenovirus serotype 4 (FAdV-4) are avian viruses that have emerged in recent years and have endangered the global poultry industry, causing great economic loss. In this study, a multiplex fluorescence-based loop-mediated isothermal amplification (mLAMP) assay for detecting ChPV, CIAV and FAdV-4 was developed to simultaneously diagnose single and mixed infections in chickens. Three primer sets and composite probes were designed according to the conserved regions of the NS gene of ChPV, VP1 gene of CIAV and hexon gene of FAdV-4. Each composite probe was labelled with a different fluorophore, which was detached to release the fluorescence signal after amplification. The target viruses were distinguished based on the colour of the mLAMP products. The mLAMP assay was shown to be sensitive, with detection limits of 307 copies of recombinant plasmids containing the ChPV target genes, 749 copies of CIAV and 648 copies of FAdV-4. The assay exhibited good specificity and no cross-reactivity with other symptomatically related avian viruses. When used on field materials, the results of the mLAMP assay were in 100% agreement with those of the previously published PCR assay. The mLAMP assay is rapid, economical, sensitive and specific, and the results of amplification are directly observable by eye. Therefore, the mLAMP assay is a useful tool for the clinical detection of ChPV, CIAV and FAdV-4 and can be applied in rural areas.


Assuntos
Infecções por Adenoviridae , Aviadenovirus , Vírus da Anemia da Galinha , Infecções por Parvoviridae , Doenças das Aves Domésticas , Animais , Adenoviridae , Infecções por Adenoviridae/diagnóstico , Infecções por Adenoviridae/veterinária , Aviadenovirus/genética , Vírus da Anemia da Galinha/genética , Galinhas/virologia , Filogenia , Doenças das Aves Domésticas/diagnóstico , Sorogrupo , Infecções por Parvoviridae/diagnóstico , Infecções por Parvoviridae/veterinária
3.
J Craniofac Surg ; 34(4): 1325-1328, 2023 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-36856445

RESUMO

BACKGROUND: Extensive facial burn scars are a tragedy for patients and often pose a great challenge to surgeons because of the high esthetic and functional demands. For patients with healthy skin in the neck region, a cervical flap is highly recommended for facial resurfacing; however, the skin on the midline of the neck often needs more expansion than that on either side, especially for the treatment of large facial defects. The sufficient longitudinal soft tissue in the anterior neck ensures a normal neck shape as well as a normal range of cervical extension, rotation, and lateral flexion. To overcome this, we developed an expanded cervical flap with an overlapping tissue expansion technique to gain more length centrally. METHODS: First, 2 tissue expanders were embedded in the anterior neck region overlapping each other at the midline of the neck. After adequate inflation of the expander, the expanded flap was dissected and rotated to repair defects in the middle and lower face. The anchor position of the flap was placed on the horizontal line of the thyroid cartilage to restore the cervicomental angle. RESULTS: Sixteen patients were treated with this method in this single-center study. All defects affected the middle and lower face, with an area ranging from 135 to 185 cm 2 , and were caused by a massive facial burn. Among them, 12 patients suffered ectropion of the lower lip, 3 suffered limited mouth opening due to scar contraction, and one patient had a cervicomental adhesion. The area of the expanded flap was approximately 163 to 266 cm 2 . The average period of expansion was 89.5 days. Patients were followed up after the operation, with the follow-up period ranging from 6 to 12 months. In all cases, good defect coverage was achieved, with primary closure of the donor sites and a good postoperative cervical configuration. CONCLUSION: We conclude that the expanded cervical flap with the overlapping tissue expansion technique proved to be a reliable method for facial skin reconstruction with functional and aesthetic improvement.


Assuntos
Queimaduras , Traumatismos Faciais , Lesões do Pescoço , Procedimentos de Cirurgia Plástica , Humanos , Transplante de Pele/métodos , Queimaduras/cirurgia , Estética Dentária , Expansão de Tecido/métodos , Cicatriz/cirurgia , Lesões do Pescoço/cirurgia , Traumatismos Faciais/cirurgia
4.
J Craniofac Surg ; 34(5): 1580-1583, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37220649

RESUMO

BACKGROUND: Traumatic injury or tumor resection can lead to eyelid defects, nasal defects, and cheek defects. The temporal flap pedicled with orbicularis oculi muscle (OOM) can be used to repair these defects. This cadaver-based anatomic study aimed to evaluate the blood supply of this flap and investigate its clinical implications. METHODS: Twenty hemifaces from 10 cadavers were used in this study. The number of arteries supplying OOM of the flap, the diameter of the artery entering OOM, and the maximum width of OOM were recorded. All data were presented as mean±SD values and analyzed using Student t -test. A P value<0.05 was considered statistically significant. RESULTS: Of these 10 specimens, 7 were males and 3 were females. The average age was 67.7 years (range, 53-78 y). The number of arteries supplying OOM was 8.5±1.4 in the male and 7.8±1.2 in the female. The diameter of the zygomatico-orbital artery was detected as 0.53±0.06 mm in the male and 0.40±0.11 mm in the female. The maximum width of OOM was detected as 2.5±0.1 cm in the male and 2.2±0.1 cm in the female. Males had significantly larger average values than females in the diameter of zygomatico-orbital artery and maximum width of OOM ( P =0.012, P <0.001, respectively). However, the number of arteries supplying OOM did not differ significantly between sex ( P =0.322). CONCLUSIONS: We conclude that the blood supply of the temporal flap pedicled with OOM is abundant and reliable. The findings provide surgeons with valuable anatomic knowledge for repairing facial defects with this flap.


Assuntos
Músculos Faciais , Retalhos Cirúrgicos , Humanos , Masculino , Feminino , Idoso , Músculos Faciais/anatomia & histologia , Pálpebras/cirurgia , Face , Bochecha
5.
Facial Plast Surg ; 2023 Oct 09.
Artigo em Inglês | MEDLINE | ID: mdl-37699518

RESUMO

Over the past 20 years, we have designed various types of expanded cervical flaps for large facial defects and achieved excellent tissue matching. This study was performed to propose a treatment strategy for flap selection for the reconstruction of different facial units. The authors retrospectively reviewed the application of cervical expanded flaps for facial rehabilitation in our department between January 2003 and January 2023. The study included 122 patients with unilateral (62.3%) and bilateral (37.7%) facial deformities ranging from the zygomatic arch to the chin. The median area of the tissue defect was 15.2 × 8.5 cm2 (ranging from 6 × 4 cm2 to 27 × 12 cm2). The expansion period ranged from 61 to 175 days (mean: 86.5 days). Maximum and minimum sizes of pre-expanded cervical flaps were 30 × 13 cm2 to 7 × 5 cm2. All the flaps could be summarized into type 1, an advanced expanded cervical flap; type 2, a wing-shaped expanded cervical flap with overlapping tissue expansion; and type 3, an expanded single-lobed transposition flap rotated based on the anterior neck. Cervical flaps reliably meet the reconstructive requirements for different facial units, especially for large cutaneous defects in the clinic. The selection of these flaps can be planned preoperatively according to the location and size of the defect or lesion.

6.
Appl Microbiol Biotechnol ; 106(11): 4211-4221, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35622123

RESUMO

A high production mutated strain Bacillus thuringiensis X023PN (BtX023PN) was screened from the wild strain Bacillus thuringiensis X023 (BtX023) after atmospheric and room temperature plasma (ARTP) and nitrosoguanidine (NTG) mutation. BtX023PN grows faster than the wild strain, and its lysis of mother cell was 6 h ahead BtX023, but the ability of sporulation was significantly reduced. Bioassay indicated that compared with the wild type strain, the virulence of BtX023PN against Plutella xylostella (P. xylostella) and Mythimna seperata (M. seperata) increased to 2.33-fold and 2.13-fold respectively. qRT-PCR and SDS-PAGE demonstrated that the production of Cry1Ac increased by 61%. Resequence indicated that the mutated sites enriched on the key carbohydrate metabolism and amino acid metabolism. This study provides a new strain resource for the development of Bt insecticides and a feasible technical strategy for the breeding of Bt. KEY POINTS: • Atmospheric and room temperature plasma used in breeding of Bacillus thuringiensis. • Less stationary phase time with more ICP production. • Semi-lethal concentration against Plutella xylostella reduced by about 57.


Assuntos
Bacillus thuringiensis , Mariposas , Animais , Bacillus thuringiensis/metabolismo , Toxinas de Bacillus thuringiensis , Proteínas de Bactérias/metabolismo , Endotoxinas/genética , Endotoxinas/metabolismo , Proteínas Hemolisinas/genética , Proteínas Hemolisinas/metabolismo , Larva , Mutação , Nitrosoguanidinas , Virulência
7.
J Craniofac Surg ; 33(7): 2220-2223, 2022 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-35765126

RESUMO

PURPOSE: To present our experience with pre-expanded medial upper arm flap in facial and neck reconstruction. PATIENTS AND METHODS: This was a retrospective study operated between January 1st, 2001 and January 1st, 2021, at the Plastic Surgery Hospital, Chinese Academy of Medical Sciences, and Peking Union Medical College. Staged face and/or neck reconstruction was performed. RESULTS: Forty-one patients were treated in our institution and thirty-eight patients (forty-three flaps) were included in this cohort as. They ranged from 6 to 44 years old. There was no total flap loss in the cohort. Partial flap necrosis was observed in the earlier patients (4 cases). CONCLUSION: Pre-expanded medial upper arm flap is well matched to the facial and neck skin in color, texture, and thickness. Considering the excellent aesthetic outcomes, this flap is a good alternative for selected patients with soft tissue defects of the head and neck.


Assuntos
Braço , Procedimentos de Cirurgia Plástica , Adolescente , Adulto , Braço/cirurgia , Criança , Estética Dentária , Humanos , Procedimentos de Cirurgia Plástica/métodos , Estudos Retrospectivos , Retalhos Cirúrgicos/cirurgia , Adulto Jovem
8.
J Craniofac Surg ; 33(8): 2543-2547, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36184764

RESUMO

OBJECTIVE: Reconstruction of facial soft-tissue defects may pose a dilemma for plastic surgeons, as the flaps must be reliable to obtain a natural appearance while minimizing donor site morbidities. This clinical study describes a reconstructive method for infraorbital and zygomatic defects using a pre-expanded rotation flap based on the orbicularis oculi muscle (OOM). METHODS: The surgeries were subdivided into 2 stages. In the first stage of the operation, a 100 to 200 mL expander was placed underneath the temporal area through a hairline incision. In the second stage, after adequate inflation of the expander, the pre-expanded rotation flap based on the OOM of the lower eyelid was raised from lateral to medial to cover the facial defects. RESULTS: In this single-center study from February 2010 to February 2017, 16 patients underwent facial defect reconstruction using the pre-expanded flap based on the OOM. All of the defects were located at the infraorbital and zygomatic regions, and their sizes ranged from 3.0 4.0 to 7.0 14.0 cm. The causes of these defects included postburn scars (37.5%), melanocytic nevus (50%), and hemangiomas (12.5%). In all cases, good coverage was provided for the defects that were in the medial cheek or lower eyelids. There were no flap losses of any kind. There were no major complications, and all minor incidences were treated by minimal procedures. The patients were followed up after surgery, with the follow up ranging from 6 months to 108 months. The follow-up data included postoperative consultations, the defect size, the need for further procedures and the degree of satisfaction. CONCLUSION: The pre-expanded rotation flaps in the lateral facial area based on the OOM can ideally and safely be applied for facial defect reconstruction owing to their reliable blood supply and excellent texture match.


Assuntos
Procedimentos de Cirurgia Plástica , Neoplasias Cutâneas , Humanos , Seguimentos , Procedimentos de Cirurgia Plástica/métodos , Retalhos Cirúrgicos/cirurgia , Músculos Faciais/cirurgia , Pálpebras/cirurgia , Neoplasias Cutâneas/cirurgia
9.
Environ Microbiol ; 23(4): 2230-2243, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33331075

RESUMO

Lysine metabolism plays an important role in the formation of the insecticidal crystal proteins of Bacillus thuringiensis (Bt). The genes lam, gabD and sucA encode three key enzymes of the lysine metabolic pathway in Bt4.0718. The lam gene mainly affects the cell growth at stable period, negligibly affected sporulation and insecticidal crystal protein (ICP) production. While, the deletion mutant strains of the gabD and sucA genes showed that the growth, sporulation and crystal protein formation were inhibited, cells became slender, and insecticidal activity was significantly reduced. iTRAQ proteomics and qRT-PCR used to analyse the differentially expressed protein (DEP) between the two mutant strains and the wild type strain. The functions of DEPs were visualized and statistically classified, which affect bacterial growth and metabolism by regulating biological metabolism pathways: the major carbon metabolism pathways, amino acid metabolism, oxidative phosphorylation pathways, nucleic acid metabolism, fatty acid synthesis and peptidoglycan synthesis. The gabD and sucA genes in lysine metabolic pathway are closely related to the sporulation and crystal proteins formation. The effects of DEPs and functional genes on basic cellular metabolic pathways were studied to provide new strategies for the construction of highly virulent insecticidal strains, the targeted transformation of functional genes.


Assuntos
Bacillus thuringiensis , Bacillus thuringiensis/genética , Proteínas de Bactérias/genética , Endotoxinas , Técnicas de Inativação de Genes , Proteínas Hemolisinas , Lisina
10.
Arch Virol ; 164(10): 2451-2458, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31273469

RESUMO

Avian reovirus (ARV) is a member of the genus Orthoreovirus in the family Reoviridae and causes a severe syndrome including viral arthritis that leads to considerable losses in the poultry industry. Innate immunity plays a significant role in host defense against ARV. Here, we explored the interaction between ARV and the host innate immune system by measuring mRNA expression levels of several genes associated with the MDA5 signaling pathway. The results showed that expression peaks for MDA5, MAVS, TRAF3, TRAF6, IRF7, IKKɛ, TBK1 and NF-κB occurred at 3 days postinfection (dpi). Moreover, type I IFN (IFN-α, IFN-ß) and IL-12 expression levels peaked at 3 dpi, while type II IFN (IFN-γ), IL-6, IL-17 and IL-18 expression reached a maximum level at 1 dpi. IL-8 changed at 5 dpi, and IL-1ß and TNF-α changed at 7 dpi. Interestingly, several key IFN-stimulated genes (ISGs), including IFITM1, IFITM2, IFITM5, Mx1 and OASL, were simultaneously upregulated and reached maximum values at 3 dpi. These data indicate that the MDA5 signaling pathway and innate immune cytokines were induced after ARV infection, which would contribute to the ARV-host interaction, especially at the early infection stage.


Assuntos
Helicase IFIH1 Induzida por Interferon/biossíntese , Linfócitos/patologia , Orthoreovirus Aviário/crescimento & desenvolvimento , Doenças das Aves Domésticas/patologia , Infecções por Reoviridae/veterinária , Transdução de Sinais , Transcriptoma , Animais , Galinhas , Citocinas/biossíntese , Interações Hospedeiro-Patógeno , Imunidade Inata
11.
Gynecol Endocrinol ; 35(6): 544-547, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30777477

RESUMO

This study was conducted to investigate the association of estrogen receptor (ER) and progesterone receptor (PR) expressions with thin endometrium. Patients with endometrial thickness of less than 7 mm were classified as the study group, while the control group was comprised of patients with endometrial thickness of 7 to 14 mm. The expressions of ER and PR were detected with semi-quantitative immunohistochemical analysis, and the differences were compared between the two groups. The expression of ER was significantly decreased (p < .05) in the stromal cells of thin endometrium during both proliferative and secretory phases as compared to those of normal endometrium. Likewise, ER expression was found to be lower in the glandular cells of thin endometrium than those of normal endometrium during proliferative phase. However, no significant differences were observed for the expression of PR in both glandular and stromal cells between the two groups. Thin endometrium was associated with reduced expression of ER in stromal cells both during proliferative and secretory phase, but in glandular epithelial cells only during proliferative phase.


Assuntos
Endométrio/metabolismo , Receptores de Estrogênio/metabolismo , Adulto , Feminino , Humanos , Histeroscopia , Imuno-Histoquímica , Ciclo Menstrual/metabolismo , Receptores de Progesterona/metabolismo , Células Estromais/metabolismo
12.
Arch Virol ; 161(8): 2243-8, 2016 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-27233800

RESUMO

The present study was conducted to identify avian reovirus (ARV) proteins that can activate the phosphatidylinositol 3-kinase (PI3K)-dependent Akt pathway. Based on ARV protein amino acid sequence analysis, σA, σNS, µA, µB and µNS were identified as putative proteins capable of mediating PI3K/Akt pathway activation. The recombinant plasmids σA-pcAGEN, σNS-pcAGEN, µA-pcAGEN, µB-pcAGEN and µNS-pcAGEN were constructed and used to transfect Vero cells, and the expression levels of the corresponding genes were quantified by immunofluorescence and Western blot analysis. Phosphorylated Akt (P-Akt) levels in the transfected cells were measured by flow cytometry and Western blot analysis. The results showed that the σA, σNS, µA, µB and µNS genes were expressed in Vero cells. σA-expressing and σNS-expressing cells had higher P-Akt levels than negative control cells, pcAGEN-expressing cells and cells designed to express other proteins (i.e., µA, µB and µNS). Pre-treatment with the PI3K inhibitor LY294002 inhibited Akt phosphorylation in σA- and σNS-expressing cells. These results indicate that the σA and σNS proteins can activate the PI3K/Akt pathway.


Assuntos
Doenças das Aves/enzimologia , Doenças das Aves/virologia , Orthoreovirus Aviário/metabolismo , Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Proteínas de Ligação a RNA/metabolismo , Infecções por Reoviridae/enzimologia , Infecções por Reoviridae/veterinária , Proteínas do Core Viral/metabolismo , Proteínas Virais Reguladoras e Acessórias/metabolismo , Animais , Doenças das Aves/genética , Chlorocebus aethiops , Regulação Viral da Expressão Gênica , Interações Hospedeiro-Patógeno , Orthoreovirus Aviário/genética , Fosfatidilinositol 3-Quinase/genética , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas de Ligação a RNA/genética , Infecções por Reoviridae/genética , Infecções por Reoviridae/virologia , Transdução de Sinais , Células Vero , Proteínas do Core Viral/genética , Proteínas Virais Reguladoras e Acessórias/genética
13.
Arch Virol ; 161(11): 3285-9, 2016 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-27503240

RESUMO

A previously unidentified chicken parvovirus (ChPV) strain, associated with runting-stunting syndrome (RSS), is now endemic among chickens in China. To explore the genetic diversity of ChPV strains, we determined the first complete genome sequence of a novel ChPV isolate (GX-CH-PV-7) identified in chickens in Guang Xi, China, and showed moderate genome sequence similarity to reference strains. Analysis showed that the viral genome sequence is 86.4 %-93.9 % identical to those of other ChPVs. Genetic and phylogenetic analyses showed that this newly emergent GX-CH-PV-7 is closely related to Gallus gallus enteric parvovirus isolate ChPV 798 from the USA, indicating that they may share a common ancestor. The complete DNA sequence is 4612 bp long with an A+T content of 56.66 %. We determined the first complete genome sequence of a previously unidentified ChPV strain to elucidate its origin and evolutionary status.


Assuntos
DNA Viral/química , DNA Viral/genética , Genoma Viral , Infecções por Parvoviridae/veterinária , Parvovirus/genética , Parvovirus/isolamento & purificação , Animais , Composição de Bases , Galinhas , China , Análise por Conglomerados , Infecções por Parvoviridae/virologia , Parvovirus/classificação , Filogenia , Doenças das Aves Domésticas/virologia , Análise de Sequência de DNA , Homologia de Sequência
14.
BMC Microbiol ; 15: 247, 2015 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-26518004

RESUMO

BACKGROUND: Duck viral pathogens primarily include the avian influenza virus (AIV) subtypes H5, H7, and H9; duck hepatitis virus (DHV); duck tembusu virus (DTMUV); egg drop syndrome virus (EDSV); duck enteritis virus (DEV); Newcastle disease virus (NDV); duck circovirus (DuCV); muscovy duck reovirus (MDRV); and muscovy duck parvovirus (MDPV). These pathogens cause great economic losses to China's duck breeding industry. RESULT: A rapid, specific, sensitive and high-throughput GeXP-based multiplex PCR assay consisting of chimeric primer-based PCR amplification with fluorescent labeling and capillary electrophoresis separation was developed and optimized to simultaneously detect these eleven viral pathogens. Single and mixed pathogen cDNA/DNA templates were used to evaluate the specificity of the GeXP-multiplex assay. Corresponding specific DNA products were amplified from each pathogen. Other pathogens, including duck Escherichia coli, duck Salmonella, duck Staphylococcus aureus, Pasteurella multocida, infectious bronchitis virus, and Mycoplasma gallisepticum, did not result in amplification products. The detection limit of GeXP was 10(3)copies when all twelve pre-mixed plasmids containing the target genes of eleven types of duck viruses were present. To further evaluate the reliability of GeXP, 150 clinical field samples were evaluated. Comparison with the results of conventional PCR methods for the field samples, the GeXP-multiplex PCR method was more sensitive and accurate. CONCLUSION: This GeXP-based multiplex PCR method can be utilized for the rapid differential diagnosis of clinical samples as an effective tool to prevent and control duck viruses with similar clinical symptoms.


Assuntos
Reação em Cadeia da Polimerase Multiplex/métodos , Doenças das Aves Domésticas/virologia , Viroses/diagnóstico , Viroses/veterinária , Vírus/isolamento & purificação , Animais , Circovirus/genética , Circovirus/isolamento & purificação , Diagnóstico Diferencial , Patos , Vírus da Influenza A/genética , Vírus da Influenza A/isolamento & purificação , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/veterinária , Reação em Cadeia da Polimerase Multiplex/veterinária , Vírus da Doença de Newcastle/genética , Vírus da Doença de Newcastle/isolamento & purificação , Doenças das Aves Domésticas/diagnóstico , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Viroses/virologia , Vírus/classificação , Vírus/genética
15.
Virol J ; 12: 226, 2015 Dec 30.
Artigo em Inglês | MEDLINE | ID: mdl-26715327

RESUMO

BACKGROUND: Immunosuppressive viruses are frequently found as co-infections in the chicken industry, potentially causing serious economic losses. Because traditional molecular biology methods have limited detection ability, a rapid, high-throughput method for the differential diagnosis of these viruses is needed. The objective of this study is to develop a GenomeLab Gene Expression Profiler Analyser-based multiplex PCR method (GeXP-multiplex PCR) for simultaneous detection of eight immunosuppressive chicken viruses. RESULTS: Using chimeric primers, eight such viruses, including Marek's disease virus (MDV), three subgroups of avian leucosis virus (ALV-A/B/J), reticuloendotheliosis virus (REV), infectious bursal disease virus (IBDV), chicken infectious anaemia virus (CIAV) and avian reovirus (ARV), were amplified and identified by their respective amplicon sizes. The specificity and sensitivity of the optimised GeXP-multiplex PCR assay were evaluated, and the data demonstrated that this technique could selectively amplify these eight viruses at a sensitivity of 100 copies/20 µl when all eight viruses were present. Among 300 examined clinical specimens, 190 were found to be positive for immunosuppressive viruses according to this novel assay. CONCLUSION: The GeXP-multiplex PCR assay is a high-throughput, sensitive and specific method for the detection of eight immunosuppressive viruses and can be used for differential diagnosis and molecular epidemiological surveys.


Assuntos
Infecções por Vírus de DNA/veterinária , Técnicas de Diagnóstico Molecular/métodos , Reação em Cadeia da Polimerase Multiplex/métodos , Doenças das Aves Domésticas/diagnóstico , Doenças das Aves Domésticas/virologia , Infecções por Vírus de RNA/veterinária , Virologia/métodos , Animais , Vírus da Anemia da Galinha , Galinhas , Infecções por Vírus de DNA/diagnóstico , Infecções por Vírus de DNA/virologia , Diagnóstico Diferencial , Ensaios de Triagem em Larga Escala , Infecções por Vírus de RNA/diagnóstico , Infecções por Vírus de RNA/virologia , Sensibilidade e Especificidade , Medicina Veterinária/métodos
16.
Virol J ; 12: 22, 2015 Feb 12.
Artigo em Inglês | MEDLINE | ID: mdl-25889592

RESUMO

BACKGROUND: Infectious arthritis in broilers represents an economic and health problem, resulting in severe losses due to retarded growth and downgrading at the slaughterhouse. The most common agents associated with cases of infectious arthritis in poultry are avian reovirus (ARV) and Mycoplasma synoviae (MS). The accurate differentiation and rapid diagnosis of ARV and MS are essential prerequisites for the effective control and prevention of these avian pathogens in poultry flocks. This study thus aimed to develop and validate a duplex real-time PCR assay for the simultaneous detection and quantification of ARV and MS. METHODS: Specific primers and probes for each pathogen were designed to target the special sequence of the ARV σC gene or the MS phase-variable surface lipoprotein hemagglutinin (vlhA) gene. A duplex real-time PCR assay was developed, and the reaction conditions were optimized for the rapid detection and quantification of ARV and MS. RESULTS: The duplex real-time PCR assay was capable of ARV- and MS-specific detection without cross-reaction with other non-targeted avian pathogens. The sensitivity of this assay was 2 × 10(1) copies for a recombinant plasmid containing ARV σC or MS vlhA gene, and 100 times higher than that of conventional PCR. This newly developed PCR assay was also reproducible and stable. All tested field samples of ARV and/or MS were detectable with this duplex real-time PCR assay compared with pathogen isolation and identification as well as serological tests. CONCLUSION: This duplex real-time PCR assay is highly specific, sensitive and reproducible and thus could provide a rapid, specific and sensitive diagnostic tool for the simultaneous detection of ARV and MS in poultry flocks. The assay will be useful not only for clinical diagnostics and disease surveillance but also for the efficient control and prevention of ARV and MS infections.


Assuntos
Artrite Infecciosa/veterinária , Reação em Cadeia da Polimerase Multiplex/métodos , Mycoplasma synoviae/isolamento & purificação , Orthoreovirus Aviário/isolamento & purificação , Doenças das Aves Domésticas/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Artrite Infecciosa/diagnóstico , Carga Bacteriana/métodos , Proteínas de Bactérias/genética , Primers do DNA/genética , Hemaglutininas/genética , Lipoproteínas/genética , Proteínas de Membrana/genética , Técnicas de Diagnóstico Molecular/métodos , Aves Domésticas , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Carga Viral/métodos , Proteínas Virais/genética
17.
Virol J ; 12: 145, 2015 Sep 17.
Artigo em Inglês | MEDLINE | ID: mdl-26377809

RESUMO

BACKGROUND: The H10 subtype avian influenza viruses (H10N4, H10N5 and H10N7) have been reported to cause disease in mammals, and the first human case of H10N8 subtype avian influenza virus was reported in 2013. Recently, H10 subtype avian influenza viruses (AIVs) have been followed more closely, but routine diagnostic tests are tedious, less sensitive and time consuming, rapid molecular detection assays for H10 AIVs are not available. METHODS: Based on conserved sequences within the HA gene of the H10 subtype AIVs, specific primer sets of H10 subtype of AIVs were designed and assay reaction conditions were optimized. A reverse-transcription loop-mediated isothermal amplification (RT-LAMP) assay was established for the rapid detection of H10 subtype AIVs. The specificity was validated using multiple subtypes of AIVs and other avian respiratory pathogens, and the limit of detection (LOD) was tested using concentration gradient of in vitro-transcribed RNA. RESULTS: The established assay was performed in a water bath at 63 °C for 40 min, and the amplification result was visualized directly as well as under daylight reflections. The H10-RT-LAMP assay can specifically amplify H10 subtype AIVs and has no cross-reactivity with other subtypes AIVs or avian pathogens. The LOD of the H10-RT-LAMP assay was 10 copies per µL of in vitro-transcribed RNA. CONCLUSIONS: The RT-LAMP method reported here is demonstrated to be a potentially valuable means for the detection of H10 subtype AIV and rapid clinical diagnosis, being fast, simple, and low in cost. Consequently, it will be a very useful screening assay for the surveillance of H10 subtype AIVs in underequipped laboratories as well as in field conditions.


Assuntos
Vírus da Influenza A/isolamento & purificação , Influenza Humana/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Amplificação de Ácido Nucleico/métodos , Zoonoses/diagnóstico , Animais , Primers do DNA/genética , Glicoproteínas de Hemaglutininação de Vírus da Influenza/genética , Humanos , Vírus da Influenza A/genética , Influenza Humana/virologia , Transcrição Reversa , Sensibilidade e Especificidade , Temperatura , Fatores de Tempo , Zoonoses/virologia
18.
Virol J ; 11: 76, 2014 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-24775810

RESUMO

BACKGROUND: Duck circovirus (DuCV) infection in farmed ducks is associated with growth problems or retardation syndromes. Rapid identification of DuCV infected ducks is essential to control DuCV effectively. Therefore, this study aims to develop of an assay for DuCV to be highly specific, sensitive, and simple without any specialized equipment. METHODS: A set of six specific primers was designed to target the sequences of the Rep gene of DuCV, and A loop-mediated isothermal amplification (LAMP) assay were developed and the reaction conditions were optimized for rapid detection of DuCV. RESULTS: The LAMP assay reaction was conducted in a 62°C water bath condition for 50 min. Then the amplification products were visualized directly for color changes. This LAMP assay is highly sensitive and able to detect twenty copies of DuCV DNA. The specificity of this LAMP assay was supported by no cross-reaction with other duck pathogens. CONCLUSION: This LAMP method for DuCV is highly specific and sensitive and can be used as a rapid and direct diagnostic assay for testing clinical samples.


Assuntos
Infecções por Circoviridae/veterinária , Circovirus/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/métodos , Doenças das Aves Domésticas/diagnóstico , Doenças das Aves Domésticas/virologia , Animais , Infecções por Circoviridae/diagnóstico , Infecções por Circoviridae/virologia , Cor , Primers do DNA/genética , DNA Viral/genética , Patos , Sensibilidade e Especificidade , Temperatura , Tempo
19.
Virus Genes ; 48(2): 381-6, 2014 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-24368760

RESUMO

Avian reovirus infection causes considerable economic loss to the commercial poultry industry. Live-attenuated vaccine strain S1133 (v-S1133, derived from parent strain S1133) is considered the safest and most effective vaccine and is currently used worldwide. To identify the genes responsible for its attenuation, DNA sequences of open reading frames (ORF) of S1133 and its parent strains S1133, 1733, 526, and C78 along with three field isolates (GuangxiR1, GuangxiR2, and GX110058) and one isolate (GX110116) from a vaccinated chicken were performed. The sequence data were compared with available sequences in nucleotide sequence databases of American (AVS-B, 138, 176) and Chinese (C-98 and T-98) origin. Sequence analysis identified that several v-S1133 specific nucleotide substitutions existed in the ORFs of λA, λB, λC, µA, µB, µNS, σA, σB, and σNS genes. The v-S1133 strain could be differentiated from the field-isolated strains based on single nucleotide polymorphisms. Phylogenetic analysis revealed that v-S1133 shared the highest sequence homologies with S1133 and reovirus isolates from China, grouped together in one cluster. Chinese isolates were clearly more distinct from the American reovirus AVS-B strain, which is associated with runting-stunting syndrome in broilers.


Assuntos
Genoma Viral , Orthoreovirus Aviário/genética , Filogenia , Fases de Leitura Aberta , Orthoreovirus Aviário/classificação , Reação em Cadeia da Polimerase , Polimorfismo de Nucleotídeo Único
20.
Front Cell Infect Microbiol ; 14: 1370414, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-38915924

RESUMO

Fowl adenovirus serotype 4 (FAdV-4) is highly pathogenic to broilers aged 3 to 5 weeks and has caused considerable economic loss in the poultry industry worldwide. FAdV-4 is the causative agent of hydropericardium-hepatitis syndrome (HHS) or hydropericardium syndrome (HPS). The virus targets mainly the liver, and HPS symptoms are observed in infected chickens. This disease was first reported in Pakistan but has now spread worldwide, and over time, various deletions in the FAdV genome and mutations in its major structural proteins have been detected. This review provides detailed information about FAdV-4 genome organization, physiological features, epidemiology, coinfection with other viruses, and host immune suppression. Moreover, we investigated the role and functions of important structural proteins in FAdV-4 pathogenesis. Finally, the potential regulatory effects of FAdV-4 infection on ncRNAs are also discussed.


Assuntos
Infecções por Adenoviridae , Aviadenovirus , Galinhas , Genoma Viral , Doenças das Aves Domésticas , Sorogrupo , Animais , Galinhas/virologia , Doenças das Aves Domésticas/virologia , Aviadenovirus/genética , Aviadenovirus/classificação , Aviadenovirus/patogenicidade , Infecções por Adenoviridae/veterinária , Infecções por Adenoviridae/virologia , Coinfecção/virologia , Coinfecção/veterinária
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