Rapid detection of Marek's disease virus in feather follicles by loop-mediated amplification. 2011

Grzegorz Woźniakowski, and Elzbieta Samorek-Salamonowicz, and Wojciech Kozdruń
Department of Poultry Viral Diseases, National Veterinary Research Institute, Partyzantów 57 Avenue, 24-100 Puławy, Poland. grzegorz.wozniakowski@piwet.pulawy.pl

Marek's disease (MD) remains a serious problem in the production of poultry. The disease is caused by Marek's disease virus (MDV), and despite the ubiquitous use of vaccination to control losses, MD still affects poultry farming worldwide. The aim of this study was to develop a loop-mediated isothermal amplification (LAMP) method for the simple and inexpensive detection of MDV in feather tips of chickens. Two pairs of specific primers complementary to the meq oncogene of MDV were designed, targeting the sequence of the very virulent MDV strain, RB1B. Bst polymerase was used for the isothermal amplification of viral DNA at 65 C for 90 min in a water bath. The fluorescence signal was identified in MDV-positive samples after the addition of SYBR Green and ultraviolet (UV) illumination. The sensitivity of LAMP was 2 log 10 plaque-forming units (PFU)/ml of HPRS16 and 10(3) copies/il of plasmid containing the target gene (meq) and was equal in sensitivity to PCR amplification. Due to the use of three sets of primers, LAMP was highly specific for MDV-1 DNA. The developed LAMP technique is a rapid and simple tool for the specific detection of MDV in samples of feathers taken from live chickens. Since the use of thermocyclers is not necessary for LAMP assay, it can be conducted by small laboratories and even field veterinarians.

UI MeSH Term Description Entries
D008380 Marek Disease A transmissible viral disease of birds caused by avian herpesvirus 2 (HERPESVIRUS 2, GALLID) and other MARDIVIRUS. There is lymphoid cell infiltration or lymphomatous tumor formation in the peripheral nerves and gonads, but may also involve visceral organs, skin, muscle, and the eye. Fowl Paralysis,Marek's Disease,Fowl Paralyses,Mareks Disease,Paralyses, Fowl,Paralysis, Fowl
D008381 Herpesvirus 2, Gallid The type species of the genus MARDIVIRUS in the family HERPESVIRIDAE. It is the etiologic agent of MAREK DISEASE, infecting domestic fowl and wild birds. Fowl Paralysis Virus,Marek's Disease Herpesvirus 1,Marek's Disease Virus Serotype 1,Neurolymphomatosis Virus,Gallid Herpesvirus 2,Herpesvirus 2 (gamma), Gallid,Marek Disease Herpesvirus 1,Fowl Paralysis Viruses,Neurolymphomatosis Viruses,Paralysis Virus, Fowl,Paralysis Viruses, Fowl
D009856 Oncogene Proteins, Viral Products of viral oncogenes, most commonly retroviral oncogenes. They usually have transforming and often protein kinase activities. Viral Oncogene Proteins,Viral Transforming Proteins,v-onc Proteins,Transforming Proteins, Viral,v onc Proteins
D002645 Chickens Common name for the species Gallus gallus, the domestic fowl, in the family Phasianidae, order GALLIFORMES. It is descended from the red jungle fowl of SOUTHEAST ASIA. Gallus gallus,Gallus domesticus,Gallus gallus domesticus,Chicken
D004279 DNA, Viral Deoxyribonucleic acid that makes up the genetic material of viruses. Viral DNA
D005241 Feathers Flat keratinous structures found on the skin surface of birds. Feathers are made partly of a hollow shaft fringed with barbs. They constitute the plumage. Feather
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D012680 Sensitivity and Specificity Binary classification measures to assess test results. Sensitivity or recall rate is the proportion of true positives. Specificity is the probability of correctly determining the absence of a condition. (From Last, Dictionary of Epidemiology, 2d ed) Specificity,Sensitivity,Specificity and Sensitivity
D021141 Nucleic Acid Amplification Techniques Laboratory techniques that involve the in-vitro synthesis of many copies of DNA or RNA from one original template. DNA Amplification Technic,DNA Amplification Technique,DNA Amplification Techniques,Nucleic Acid Amplification Technic,Nucleic Acid Amplification Technique,RNA Amplification Technic,RNA Amplification Technique,RNA Amplification Techniques,Amplification Technics, Nucleic Acid,Amplification Techniques, Nucleic Acid,DNA Amplification Technics,Nucleic Acid Amplification Technics,Nucleic Acid Amplification Test,Nucleic Acid Amplification Tests,RNA Amplification Technics,Technics, Nucleic Acid Amplification,Techniques, Nucleic Acid Amplification,Amplification Technic, DNA,Amplification Technic, RNA,Amplification Technics, DNA,Amplification Technics, RNA,Amplification Technique, DNA,Amplification Technique, RNA,Amplification Techniques, DNA,Amplification Techniques, RNA,Technic, DNA Amplification,Technic, RNA Amplification,Technics, DNA Amplification,Technics, RNA Amplification,Technique, DNA Amplification,Technique, RNA Amplification,Techniques, DNA Amplification,Techniques, RNA Amplification
D025202 Molecular Diagnostic Techniques MOLECULAR BIOLOGY techniques used in the diagnosis of disease. Molecular Testing,Molecular Diagnostic Technics,Molecular Diagnostic Testing,Diagnostic Technic, Molecular,Diagnostic Technics, Molecular,Diagnostic Technique, Molecular,Diagnostic Techniques, Molecular,Diagnostic Testing, Molecular,Molecular Diagnostic Technic,Molecular Diagnostic Technique,Technic, Molecular Diagnostic,Technics, Molecular Diagnostic,Technique, Molecular Diagnostic,Techniques, Molecular Diagnostic,Testing, Molecular,Testing, Molecular Diagnostic

Related Publications

Grzegorz Woźniakowski, and Elzbieta Samorek-Salamonowicz, and Wojciech Kozdruń
March 2012, Journal of clinical microbiology,
Grzegorz Woźniakowski, and Elzbieta Samorek-Salamonowicz, and Wojciech Kozdruń
April 2005, Journal of clinical microbiology,
Grzegorz Woźniakowski, and Elzbieta Samorek-Salamonowicz, and Wojciech Kozdruń
August 2012, Journal of virological methods,
Grzegorz Woźniakowski, and Elzbieta Samorek-Salamonowicz, and Wojciech Kozdruń
March 2020, Journal of virological methods,
Grzegorz Woźniakowski, and Elzbieta Samorek-Salamonowicz, and Wojciech Kozdruń
September 2015, Virusdisease,
Grzegorz Woźniakowski, and Elzbieta Samorek-Salamonowicz, and Wojciech Kozdruń
August 1970, Archivio veterinario italiano,
Grzegorz Woźniakowski, and Elzbieta Samorek-Salamonowicz, and Wojciech Kozdruń
October 2010, Journal of virological methods,
Grzegorz Woźniakowski, and Elzbieta Samorek-Salamonowicz, and Wojciech Kozdruń
February 2019, Virus genes,
Grzegorz Woźniakowski, and Elzbieta Samorek-Salamonowicz, and Wojciech Kozdruń
January 2011, Microbiology and immunology,
Grzegorz Woźniakowski, and Elzbieta Samorek-Salamonowicz, and Wojciech Kozdruń
November 2018, Journal of virological methods,
Copied contents to your clipboard!