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

2007 ◽  
Vol 19 (5) ◽  
pp. 471-478 ◽  
Author(s):  
Shiro Murata ◽  
Kyung-Soo Chang ◽  
Sung-Il Lee ◽  
Satoru Konnai ◽  
Misao Onuma ◽  
...  

For the easy survey of Marek's disease virus (MDV), feather tip–derived DNA from MDV-infected 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.

2019 ◽  
Vol 35 (1-2) ◽  
pp. 1-6
Author(s):  
MW Rahman ◽  
M Nooruzzaman ◽  
US Suma ◽  
EH Chowdhury ◽  
MR Islam

A total of 24 clinical specimens (10 feathers, 8 peripheral bloods and 6 spleens) were collected from 10 suspected outbreaks of Marek’s disease (MD) from Mymensingh, Tangail, Gazipur and Pabna districts of Bangladesh. A polymerase chain reaction (PCR) protocol originally described by Silva (1992) was adopted to detect Marek’s disease virus (MDV) genome in these specimens. All the tested peripheral blood buffy coat samples (100%) were positive for MDV in PCR, while 70% of feather samples and 66.6% of spleen samples were positive. A band of 317 bp size was found in all positive samples. A few samples also yielded additional bands of 185 bp size and/or multiple bands of larger than 317 bp size, indicating the presence of both virulent MDV and the vaccine virus. The study suggests that peripheral blood and feathers from live birds, and feathers from dead birds are the samples of choice for the detection of MDV by PCR. The Bangladesh Veterinarian (2018) 35(1&2): 1-6


1992 ◽  
Vol 36 (3) ◽  
pp. 637 ◽  
Author(s):  
Geng-Sheng Zhu ◽  
Tomoko Ojima ◽  
Takashi Hironaka ◽  
Takeshi Ihara ◽  
Noriko Mizukoshi ◽  
...  

2021 ◽  
pp. 1342-1353
Author(s):  
Nahed Yehia ◽  
Hemat S. El-Sayed ◽  
Sabry E. Omar ◽  
Ahmed Erfan ◽  
Fatma Amer

Background and Aim: The Marek's disease virus (MDV) is a neoplastic disease causing serious economic losses in poultry production. This study aimed to investigate MDV occurrence in poultry flocks in the Lower Egypt during the 2020 breakout and genetically characterized Meq, gL, and ICP4 genes in field strains of MDV. Materials and Methods: Forty samples were collected from different breeds from eight Egyptian governorates in 2020. All flocks had received a bivalent vaccine (herpesvirus of turkey FC-126 + Rispens CVI988). However, weight loss, emaciation, reduced egg production, paralysis, and rough/raised feather follicles occurred. Samples were collected from feather follicles, liver, spleen, and nerve tissue for diagnosis by polymerase chain reaction. MDV genetic characterization was then performed by sequencing the Meq, gL, and ICP4 genes of five positive samples representing different governorates and breeds. Results: A total of 28 samples were positive for MDV field strains, while two were related to MDV vaccinal strains. All samples tested negative for ALV (A, B, C, D, and J) and REV. Phylogenetic analysis of the Meq gene of sequenced samples revealed that all MDVs were related to the highly virulent European viruses (Gallid herpesvirus 2 ATE and PC12/30) with high amino acid (A.A.) identity 99.2-100%. Alternatively, there was low A.A. identity with the vaccine strains CVI988 and 3004 (up to 82.5%). These results indicate that further investigation of the efficacy of current Egyptian vaccines is required. The Egyptian strains also harbor a specific mutation, allowing clustering into two subgroups (A and B). By mutation analysis of the Meq gene, the Egyptian viruses in our study had R101K, P217A, and E263D mutations present in all Egyptian viruses. Furthermore, R176A and T180A mutations specific to our strains contributed to the high virulence of highly virulent strains. There were no mutations of the gL or ICP4 genes. Conclusion: Further studies should evaluate the protection contributed by current vaccines used in Egypt.


VirusDisease ◽  
2018 ◽  
Vol 29 (2) ◽  
pp. 167-172 ◽  
Author(s):  
Kekungu-u Puro ◽  
Uttaran Bhattacharjee ◽  
Samprity Baruah ◽  
Arnab Sen ◽  
Samir Das ◽  
...  

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