Marek's disease virus genome separation from feather tip extracts by pulsed field gel electrophoresis

Borenshtain, Rinat; Davidson, Irit

doi:10.1016/s0166-0934(01)00434-7

Cited by 15 publications

(14 citation statements)

References 14 publications

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“…11 To verify whether this difference in detection Figure 1. Nested PCR amplification of the meq gene from A, feather tips, or B, PBMC of chickens infected with 1,000 PFU/ chicken of strain Md5 (1, 2); 2,000 PFU/chicken of strain CVI988 (3, 4); 1,000 PFU/chicken of strain FC126 (5, 6); 1,000 PFU/chicken of Md5 and 2,000 PFU/chicken of CVI988 (7,8); or 1,000 PFU/chicken of Md5 and 2,000 PFU/chicken of FC-126 (9, 10). DNA samples were collected at 5 weeks pi.…”

Section: Resultsmentioning

confidence: 99%

Development of a Nested Polymerase Chain Reaction Method to Detect Oncogenic Marek's Disease Virus from Feather Tips

et al. 2007

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Abstract. For the easy survey of Marek's disease virus (MDV), feather tip-derived DNA from MDVinfected chickens can be used because feather tips are easy to collect and feather follicle epithelium is known to be the only site of productive replication of cell-free MDV. To develop a diagnostic method to differentiate highly virulent strains of MDV from the attenuated MDV vaccine strain, CVI988, which is widely used, nested polymerase chain reaction (PCR) was performed to detect a segment of the meq gene in feather tip samples of chickens experimentally infected with MDV. In chickens infected with Md5, a strain of oncogenic MDV, the meq gene was consistently detected, whereas the L-meq gene, in which a 180-base pair (180-bp) sequence is inserted into the meq gene, was detected in CVI988-infected chickens. Moreover, the meq gene was mainly detected even in chickens co-infected with both Md5 and CVI988. These results suggest that this method is appropriate for the surveillance of the highly virulent MDV infection in the field.

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Section: Resultsmentioning

confidence: 99%

Development of a Nested Polymerase Chain Reaction Method to Detect Oncogenic Marek's Disease Virus from Feather Tips

et al. 2007

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“…DNA extracted from visceral organs of tumor‐bearing chickens, or from feather tip extracts of MDV‐infected chickens were separated by PFGE, blotted and hybridized to a MDV probe [24]. A total of 54 samples of organ origin were assayed, and in 15 of them a DNA that was identified by the probe at the gel origin, did not migrate to the 200 kbp area of the gel.…”

Section: Resultsmentioning

confidence: 99%

Novel applications of feather tip extracts from MDV-infected chickens; diagnosis of commercial broilers, whole genome separation by PFGE and synchronic mucosal infection

Davidson¹,

Borenshtain²

2003

FEMS Immunology &Amp; Medical Microbiology

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Marek's disease virus (MDV) productive replication occurs in the feather follicle epithelium and the feather tips are valuable both for research and disease diagnosis. Three novel applications of feather tip extracts are described now: (A). As a source of DNA for amplifying either MDV and/or ALV-J. In two clinical situations a marked advantage was obtained compared to blood and organs; in broiler breeder flocks with a mixed MDV and ALV-J infection, and in young broilers with neurological Marek's disease (MD). (B). Separation of the large ( approximately 200 kbp) MDV genome directly from the infected chickens. Using pulsed field gel electrophoresis, the DNA extracted from tumors or feather tips was separated and hybridized to a 132 bp tandem repeat MDV probe. Compared to 2/55 polymerase chain reaction (PCR) positive tumor samples, 15/61 feather tip extracts contained whole MDV genomes. (C). Experimental MDV infection was induced by the mucosal route by dripping feather tip extract to the eye and mouth of the bird. That attempted to reproduce the native infection process, however the use of extracts, instead of dry feather dust was a compromise, aimed to synchronize the infection. In one trial, tumors were induced 6 weeks after dripping day-old broilers, while in another, feather tips were PCR positive 16 days after dripping of 2-month-old layers.

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“…Vaccinated chickens are protected from the occurrence of clinical symptoms but not from infection, and can still shed the virus through FFE. Moreover, the continued increase in MDV virulence has prompted concern that currently available vaccines could later lose efficacy in controlling MD (8). To solve this problem, it is necessary to clarify the protective mechanism of MD vaccine.…”

mentioning

confidence: 99%

Cytokine profiles in chickens infected with virulent and avirulent Marek's disease viruses: Interferon‐gamma is a key factor in the protection of Marek's disease by vaccination

Kano

Konnai

Onuma

et al. 2009

Microbiology and Immunology

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Marek's disease has been controlled by vaccination with avirulent strains of MDV. However, the protection mechanism following vaccination is not fully understood. In this study immune responses of PBMC and splenocytes derived from vaccinated chickens challenged with virulent MDV were examined using realtime PCR and ELISA. Higher levels of IFN-γ induction were observed in chickens vaccinated during the latent phase of infection with virulent MDV than in similarly challenged, unvaccinated chickens. Furthermore, the mean expression of IFNGR2 and IFN regulatory factor-3 mRNAs was significantly higher in vaccinated than in unvaccinated chickens. These results show that IFN-γ could be one of the important factors in prevention of MD by vaccination and is effective during the latent phase of the infection.

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Marek's disease virus genome separation from feather tip extracts by pulsed field gel electrophoresis

Cited by 15 publications

References 14 publications

Development of a Nested Polymerase Chain Reaction Method to Detect Oncogenic Marek's Disease Virus from Feather Tips

Development of a Nested Polymerase Chain Reaction Method to Detect Oncogenic Marek's Disease Virus from Feather Tips

Novel applications of feather tip extracts from MDV-infected chickens; diagnosis of commercial broilers, whole genome separation by PFGE and synchronic mucosal infection

Cytokine profiles in chickens infected with virulent and avirulent Marek's disease viruses: Interferon‐gamma is a key factor in the protection of Marek's disease by vaccination

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