scholarly journals Comparative suitability of the feathers, peripheral blood and spleen tissue samples of chickens for the detection of Marek’s disease virus by polymerase chain reaction

2019 ◽  
Vol 35 (1-2) ◽  
pp. 1-6
Author(s):  
MW Rahman ◽  
M Nooruzzaman ◽  
US Suma ◽  
EH Chowdhury ◽  
MR Islam
Keyword(s):  
Polymerase Chain Reaction ◽  
Disease Virus ◽  
Peripheral Blood ◽  
Buffy Coat ◽  
Marek's Disease Virus ◽  
Marek's Disease ◽  
Marek’S Disease Virus ◽  
Marek’S Disease ◽  
Chain Reaction ◽  
Polymerase Chain

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

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 ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Disease Virus ◽  
Marek's Disease Virus ◽  
Marek's Disease ◽  
Marek’S Disease Virus ◽  
Chain Reaction ◽  
Nested Polymerase Chain Reaction ◽  
Polymerase Chain ◽  
Highly Virulent ◽  
Meq Gene

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.

Differentiation of Oncogenic and Nononcogenic Strains of Marek's Disease Virus Type 1 by Using Polymerase Chain Reaction DNA Amplification

1992 ◽  
Vol 36 (3) ◽  
pp. 637 ◽  
Author(s):  
Geng-Sheng Zhu ◽  
Tomoko Ojima ◽  
Takashi Hironaka ◽  
Takeshi Ihara ◽  
Noriko Mizukoshi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Disease Virus ◽  
Virus Type ◽  
Dna Amplification ◽  
Marek's Disease Virus ◽  
Marek's Disease ◽  
Chain Reaction ◽  
Disease Virus Type ◽  
Polymerase Chain

Isolation and identification of Marek’s disease virus (MDV) from feather follicle epithelium and internal organs of diseased chickens in Dakahlia Governorate, Egypt

2019 ◽  
Vol 20 (2) ◽  
pp. 6-11
Author(s):  
Aly El-Kenawy ◽  
Mohamed El-Tholoth ◽  
Emad A
Keyword(s):  
Real Time ◽  
Disease Virus ◽  
Real Time Pcr ◽  
Marek's Disease Virus ◽  
Marek's Disease ◽  
Marek’S Disease Virus ◽  
Marek’S Disease ◽  
Field Samples ◽  
Feather Follicle ◽  
Positive Results

In the present study, a total of 16 samples including feather follicle epithelium, ovary, spleen and kidney (4 samples for each organ) were collected from diseased chicken flocks suspected to be infected with Marek’s disease virus (MDV) at Dakahlia Governorate, Egypt during the period from October 2016 to October 2017. Each sample was pooled randomly from three to five birds (90 to 360 days old). The isolation of the suspected virus from the collected samples was carried out via chorioallantoic membranes (CAMs) of 12 days old embryonated chicken eggs (ECEs). Three egg passages were carried out for each sample. Hyperimmune serum was prepared against standard MDV. MDV in both field and egg passaged samples (after 3rd passage) was identified by agar gel precipitation test (AGPT) and indirect fluorescence antibody test (IFAT). Molecular identification of virus was carried out by conventional polymerase chain reaction (PCR) and real- time PCR in four selected samples. The results revealed that 14 samples (87.5%) including 4 (100%) samples from feather follicle epithelium, ovary and kidney and 2 (50%) samples from spleen, showed positive results in virus isolation after 3rd passage. The positive results percentage by AGPT for field samples were 50% (8 out of 16 samples), while after the 3rd passage in ECEs were 37.5% (6 out of 16 samples) and the positive results percentage by IFAT for field samples were 62.5% (10 out of 16 samples), while after the 3rd passage in ECEs were 81.25 % (13 out of 16 samples). Viral nucleic acid was detected in all selected samples by conventional and real- time PCR. The results indicate that feather follicle epithelium is the best organ for MDV detection. IFAT is superior over AGPT in virus detection. Conventional and real - time PCR could be efficiently used for molecular detection of the virus.

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