The use of serotype 1- and serotype 3-specific polymerase chain reaction for the detection of Marek's disease virus in chickens

Abstract

A serotype 1- and serotype 3-specific detection of Marek's disease virus (MDV) by polymerase chain reaction (PCR) was developed. The sensitivity of the method when applied to cell culture grown virus was comparable with that of cultivation. The method was applied to various tissue samples from chickens experimentally inoculated with serotype 1 or serotype 3 MDV.The serotype 1 strains CVI988 and RB-1B could be detected in feather follicle epithelium up to 56 and 84 days post-inoculation (p.i.), respectively, while the MDV-3 serotype was detected until 42 days p.i. The purpose of this study was to develop and evaluate a reliable and easy-to-handle method for surveillance of the occurrence of MDV in chicken flocks. We emphasize the development of a method, which can be applied to types of samples conveniently collected in the field, e.g. feather tips and blood samples. In addition, the PCR was applied to samples collected from four commercial table egg layer flocks of young stock or pullets vaccinated with either serotype 1 (CVI988) or serotype 3 (HVT) vaccine. These flocks had various clinical signs of Marek's disease. MDV-1 was detected in buffy-coat cells, spleen, liver, skin, feather tips and ovaries. The detection of MDV in feather tips appeared to be as sensitive as co-cultivation of buffy-coat cells, although an inhibiting factor was observed in extracts from feather tips of non-white chickens. This inhibition could be overcome in most extracts by applying a bovine serum albumen pretreatment. The PCR proved to be a convenient tool for the monitoring of MDV in the poultry population, and feather tips were the most convenient and sensitive samples.

Detection of Avian Leukosis Virus Subgroup J from Commercial Peking Duck Breeder Farm in Egypt

“…The ALV-J and ALV-A (Smith et al, 1998), MDV (Handberg et al, 2001) and REV (Wei et al, 2012), were performed, respectively. Primers were utilized in a 25 μL reaction containing 12.5 μL of Emerald Amp Max PCR Master Mix (Takara, Japan), 1 μL of each primer of 20 pmol concentrations, 4.5 μL of water and 6 μL of DNA template.…”

Innovation and discovery: The application of nucleic acid-based technology to avian virus detection and characterization

“…The authors suggested that the low level of gene expression of ALV-J in the bursa might help to explain the inability of ALV-J to induce lymphoid leukosis. Handberg et al (2001) have developed two PCRs, one specific for type 1 herpesviruses (pathogenic, oncogenic Marek's disease virus (MDV-1)), the other for type 3 avian herpesvirus (herpesvirus of turkeys, often used as a vaccine against Marek's disease). They were particularly interested in developing protocols for the detection and differentiation of these viruses that would be convenient for people collecting material in the field.…”

Marek's disease in broiler chickens: Effect of route of infection and herpesvirus of turkey-vaccination status on detection of virus from blood or spleen by polymerase chain reaction, and on weights of birds, bursa and spleen

“…Virus isolation is usually by virus propagation in cell culture and identificatio n/quantification by cytopathic changes (plaque formation) or identificatio n of infected cells by immunostaining (De Laney et al, 1998). Polymerase chain reaction (PCR) has emerged in recent years as an additional diagnostic tool offering the advantages of serotype specificity (Davidson et al, 1995;Walkden-Brown et al, 2001) and the ability to differentiate between vaccinal and field strains of MDV serotype-1 (Silva, 1992;Zhu et al, 1992;Handberg et al, 2001).…”