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Avian Marek's disease virus antibody ELISA kit Serum

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[#LSY-30016] Avian Marek's disease virus antibody ELISA kit Serum

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LSY-30016 | Avian Marek's disease virus antibody ELISA kit Serum, 96 wells/kit
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(1) Isolation and characterization of subgroup J Avian Leukosis virus associated with hemangioma in commercial Hy-Line chickens.[TOP]

Pubmed ID :29788333
Publication Date : //
There was an outbreak of hemangioma associated with avian leukosis virus subgroup J (ALV-J) between 2006 and 2010 in China in commercial layer chickens. Recently, severe hemangiomas broke out in Hy-Line layer chickens on a poultry farm in 2017 where ALV was eradicated earlier. Six isolates of ALV-J, named SDAU1701-SDAU1706, were characterized by virus isolation and sequence analysis of the complete proviral genomes. Avian leukosis virus subgroup J was identified by an immunofluorescence assay with monoclonal antibody JE9, whereas Marek's disease virus or reticuloendotheliosis virus was not detected. Sequence analysis of the complete proviral genome revealed that there was 96.0-99.6% identity between each other and had a homology of 94.6-96.0% when compared with the reference strain. The six isolates formed one distinct lineage separate from the reference sequences in a phylogenetic-tree, which suggested that there were several genetic differences between these groups. Homology analysis of the env, pol, and gag genes of the six isolates showed that the env gene was more variable, especially the gp85 protein, which shared only 88.2-91.9% identity with the reference strains. Sequence comparisons of the gp85 protein indicated that 19 sites were different from those in the NX0101 and HPRS-103 strains inducing myeloid leukosis; among our strains, five mutations were identical to those in the viruses causing hemangioma. Four other distinctive mutations were detected in our six isolates. This study reminds us that the surveillance of viral eradication should be conducted continuously on a farm where ALVs were eradicated. To prevent the prevalence of ALVs, more attention should be paid to daily monitoring.

Authors : Meng Fanfeng, Li Qiuchen, Zhang Yubiao, Cui Zhizhong, Chang Shuang, Zhao Peng,



(2) Synergistic Viral Replication of Marek's Disease Virus and Avian Leukosis Virus Subgroup J is Responsible for the Enhanced Pathogenicity in the Superinfection of Chickens.[TOP]

Pubmed ID :29783672
Publication Date : //
Superinfection of Marek's disease virus (MDV) and avian leukosis virus subgroup J (ALV-J) causes lethal neoplasia and death in chickens. However, whether there is synergism between the two viruses in viral replication and pathogenicity has remained elusive. In this study, we found that the superinfection of MDV and ALV-J increased the viral replication of the two viruses in RNA and protein level, and synergistically promoted the expression of IL-10, IL-6, and TGF-β in chicken embryo fibroblasts (CEF). Moreover, MDV and ALV-J protein expression in dual-infected cells detected by confocal laser scanning microscope appeared earlier in the cytoplasm and the nucleus, and caused more severe cytopathy than single infection, suggesting that synergistically increased MDV and ALV-J viral-protein biosynthesis is responsible for the severe cytopathy. In vivo, compared to the single virus infected chickens, the mortality and tumor formation rates increased significantly in MDV and ALV-J dual-infected chickens. Viral loads of MDV and ALV-J in tissues of dual-infected chickens were significantly higher than those of single-infected chickens. Histopathology observation showed that more severe inflammation and tumor cells metastases were present in dual-infected chickens. In the present study, we concluded that synergistic viral replication of MDV and ALV-J is responsible for the enhanced pathogenicity in superinfection of chickens.

Authors : Zhou Jing, Zhao Guo-Liang, Wang Xiao-Man, Du Xu-Sheng, Su Shuai, Li Chen-Gui, Nair Venugopal, Yao Yong-Xiu, Cheng Zi-Qiang,



(3) Use of real time PCR to rule out Marek's disease in the diagnosis of peripheral neuropathy.[TOP]

Pubmed ID :29745244
Publication Date : //
This article reports nine cases of neurological disease in brown layer pullets that occur in various European countries between 2015 and 2018. In all cases, the onset of neurological clinical signs was at 4-8 weeks of age and they lasted up to 22 weeks of age. Enlargement of peripheral nerves was the main lesion observed in all cases. Histopathological evaluation of nerves revealed edema with moderate to severe infiltration of plasma cells. Marek's disease (MD) was ruled out by real time PCR as none of the evaluated tissues had high load of oncogenic MD virus (MDV) DNA, characteristics of MD. Based on the epidemiological data (layers with clinical signs starting at 5-8 weeks of age), gross lesions (peripheral nerves enlargements with lack of tumors in other organs), histopathological lesions (edema and infiltration of plasma cells), and no evidence of high load of MDV DNA, we concluded that those cases were due to peripheral neuropathy (PN). PN is an autoimmune disease easily misdiagnosed as MD, leading to a costly enforcement of the vaccination protocol. Additional vaccination against MD does not protect against PN and could worsen the clinical signs by over-stimulating the immune system. Differential diagnosis between PN and MD should always be considered in cases of neurological disease with enlargement of peripheral nerves as the only gross lesion. This case report shows for the first time how real time PCR to detect oncogenic MDV is a very valuable tool in the differential diagnosis of PN and MD.

Authors : Gall Sesny, Kőrösi László, Cortes Aneg L, Delveccio Andrea, Prandini Francesco, Mitsch Peter, Gimeno Isabel M,



(4) Central Nervous System B-cell Lymphoma in a Bald Eagle ( Haliaeetus leucocephalus).[TOP]

Pubmed ID :29698076
Publication Date : //
An adult bald eagle ( Haliaeetus leucocephalus) presented for nystagmus and an inability to fly. On physical examination, the eagle was open-mouth breathing and tachycardic at 200 beats per minute, had a wrinkled cere and sunken eyes, and was an estimated 10% dehydrated. Additionally, the eagle was extremely weak, with neurologic abnormalities including bilateral proprioceptive deficits, nystagmus, and no pupillary light reflex in the left eye. Despite aggressive treatment, the eagle continued to decline rapidly and subsequently died. On histologic examination, diffuse and widespread infiltration of neoplastic lymphocytes was present in the brain, optic nerves, and pecten. Immunohistochemical PAX-5 labeling confirmed B-cell lymphoma confined to the eye and nervous system. Test results for select avian retroviruses, Marek's disease, West Nile virus, avian influenza viruses, and Mycoplasma were negative. To our knowledge, this is the first report of B-cell lymphoma in a bald eagle. Although rare, this condition is a differential diagnosis in cases of neurologic or ocular diseases in birds.

Authors : Visser Marike, Walz Heather, Shrader Stephanie, Koehler Jey, Bellah Jamie,



(5) Development and application of a SYBR green real-time PCR for detection of the emerging avian leukosis virus subgroup K.[TOP]

Pubmed ID :29617900
Publication Date : //
Avian leukosis virus subgroup K (ALV-K) is an emerging ALV tumor virus of chickens. We developed a SYBR green-based real-time polymerase chain reaction (PCR) assay for the rapid and economical detection of ALV-K in chicken flocks. The assay was specific for ALV-K and did not cross-react with other ALV subgroup or avian influenza virus, Newcastle disease virus, or Marek's Disease virus. The method was 100 times more sensitive than conventional PCR and 10 times more sensitive than the enzyme-linked immunosorbent assay (ELISA) for the P27 antigen. The assay was also more sensitive than conventional PCR in tests of 86 clinical plasma samples. DF-1 tissue culture cells infected with 1 TCID50 ALV-K particle were identified as negative using ELISA but tested positive with the real-time PCR method. The viral loads in organs and tissues in infected chickens were highest in kidney, lungs, and glandular stomach, and these results matched ELISA findings.

Authors : Chen Jian, Zhao Zijun, Chen Yangyijun, Zhang Jie, Yan Lifu, Zheng Xiaocui, Liao Ming, Cao Weisheng,



(6) First molecular detection and characterization of Marek's disease virus in red-crowned cranes (Grus japonensis): a case report.[TOP]

Pubmed ID :29615025
Publication Date : //
Marek's disease virus (MDV) resides in the genus Mardivirus in the family Herpesviridae. MDV is a highly contagious virus that can cause neurological lesions, lymphocytic proliferation, immune suppression, and death in avian species, including Galliformes (chickens, quails, partridges, and pheasants), Strigiformes (owls), Anseriformes (ducks, geese, and swans), and Falconiformes (kestrels).

Authors : Lian Xue, Ming Xin, Xu Jiarong, Cheng Wangkun, Zhang Xunhai, Chen Hongjun, Ding Chan, Jung Yong-Sam, Qian Yingjuan,



(7) Development of reliable techniques for the differential diagnosis of avian tumour viruses by immunohistochemistry and polymerase chain reaction from formalin-fixed paraffin-embedded tissue sections.[TOP]

Pubmed ID :29533078
Publication Date : //
A variety of techniques have been developed as diagnostic tools for the differential diagnosis of tumours produced by Marek's disease virus from those induced by avian leukosis virus and reticuloendotheliosis virus. However, most current techniques are unreliable when used in formalin-fixed paraffin-embedded (FFPE) tissues, which often is the only sample type available for definitive diagnosis. A collection of tumours was generated by the inoculation of different strains of Marek's disease virus, reticuloendotheliosis virus or avian leukosis virus singularly or in combination. FFPE tissue sections from tumour and non-tumour tissues were analysed by optimized immunohistochemistry (IHC) techniques and traditional as well as quantitative polymerase chain reaction (PCR) with newly designed primers ideal for DNA fragmented by fixation. IHC and PCR results were highly sensitive and specific in tissues from single-infected birds. Virus quantity was higher in tumours compared to non-tumour spleens from Marek's disease (MD) virus-infected birds. Thus, using FFPE sections alone may be sufficient for the diagnosis of MD by demonstration of high quantities of viral antigens or genome in tumour cells, along with the absence of other tumour viruses by traditional PCR, and if standard criteria are met based on clinical history and histology. IHC furthermore allowed detection of the specific cells that were infected with different viruses in tumours from birds that had been inoculated simultaneously with multiple viruses. Following validation with field samples, these new protocols can be applied for both diagnostic and research purposes to help accurately identify avian tumour viruses in routine FFPE tissue sections.

Authors : Ahmed Husnain, Mays Jody, Kiupel Matti, Dunn John R,



(8) Loop-mediated isothermal amplification assay for detection of four immunosuppressive viruses in chicken.[TOP]

Pubmed ID :29476761
Publication Date : //
Loop-mediated isothermal amplification (LAMP) methods to detect chicken infectious anemia virus (CIAV), reticuloendotheliosis virus (REV), and Marek's disease virus (MDV), and a reverse transcription (RT)-LAMP assay to detect infectious bursal disease virus (IBDV), were developed. The CIAV-LAMP, REV-LAMP, MDV-LAMP, and IBDV-RT-LAMP methods were performed using four sets of six primers targeting the VP1 gene of CIAV, the gp90 gene of REV, the Meq gene of MDV, and the VP2 gene of IBDV. The results (a change in color) were observed visually. The methods showed high specificity and sensitivity. The detection limits were 50 genomic copies of CIAV, 16 genomic copies of REV, 20 genomic copies of MDV, and 250 genomic copies of IBDV. When used to test clinical samples, the results of the LAMP assays were in 100% agreement with a previously described PCR. Therefore, the LAMP assays are simple, rapid, highly sensitive, and specific methods for detecting four immune-suppressive viruses.

Authors : Song HyeSoon, Bae YouChan, Park SeokChan, Kwon HyukMan, Lee HeeSoo, Joh SeongJoon,



(9) Design of miRNA sponges for MDV-1 as a therapeutic strategy against lymphomas.[TOP]

Pubmed ID :29423087
Publication Date : //
Lymphomas are solid-type tumors containing lymphoid cells. Some of latent herpesvirus infections established in B and/or T-lymphocytes could result in the formation of lymphomas. Marek's disease virus serotype 1 (MDV-1) is an avian herpes virus causing to lymphoproliferative tumors in birds, known as Marek's disease (MD). MD has often been used as an ideal biological model for studying the pathogenesis of lymphoma diseases caused by viruses. Therefore, we used it as a research subject to study the effect of miRNA sponges on its tumorigenicity, and to develop the theoretical basis for a new anti-tumor small molecule. The miRNA sponges designed in this study specifically bind to and degrade the miRNAs of meq gene cluster of MDV-1, including miR-M2-3p, miR-M3-5p, miR-M5-3p, miR-M9-5p and miR-M12-3p.qPCR results showed that the knockdown efficiency was 85.03%, 74.97%, 47.06%, 75.33% and 62.55%, respectively. EDU staining and CCK-8 results showed that miRNA sponges inhibited the proliferation of MDV-1 transformed MSB-1 cells , and the proliferation rate of miRNA sponges-treated cells was about 50% of the control group. DAPI staining and Annxin V-FITC/PI double staining showed that miRNA sponges induced apoptosis in MSB-1 cells, and the apoptotic rate was increased by about 27.87% compared with the control group. The results of transwell showed that miRNA sponges could inhibit the invasion of MSB-1 cells , and the inhibitory rate was about 64.52%. The soft agar assay showed that miRNA sponges could inhibit the tumorigenic ability of MSB-1 cells , and the inhibitory rate was about 66.44%.The 60-days animal study showed that miRNA sponges could alleviate the growth inhibition of MSB-1 cells (about 14.78%) and reduce the mortality (about 16.00%). In addition, the tumor formation rate was 0 (8-12% in the control group).This study suggests that miRNA sponges can serve as an effective anti-tumor small molecule for the tumors caused by herpesvirus, with potential clinical implications.

Authors : Fang Yuan, Zhou Yuqi, Zhang Yun, He Liangliang, Xue Chunyi, Cao Yongchang,



(10) Attenuation of Marek's disease virus by codon pair deoptimization of a core gene.[TOP]

Pubmed ID :29407380
Publication Date : //
Marek's disease virus (MDV) is an oncogenic alphaherpesvirus of Gallus gallus, the domesticated chicken. Control strategies rely upon vaccination with live attenuated viruses of antigenically similar avian herpesviruses or attenuated strains of MDV. Recent studies in other viruses have shown that recoding certain viral genes to employ synonymous but rarely-used codon pairs resulted in viral attenuation. We deoptimized two MDV proteins, UL54/ICP27 and UL49/VP22, and demonstrate that the more severely deoptimized variant of UL54 accumulates significantly less gene product in vitro. Using these UL54 deoptimized mutants, we further demonstrate that animals infected with the UL54-recoded recombinant virus exhibited decreased viral genome copy number in lymphocytes, reduced lymphoid atrophy and reduced tumor incidence. This study demonstrates that codon pair deoptimization of a single viral gene can produce attenuated strains of MDV. This approach may be useful as a rational way of making novel live attenuated virus vaccines for MDV.

Authors : Conrad Steven J, Silva Robert F, Hearn Cari J, Climans Megan, Dunn John R,