+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Reverse restriction fragment length polymorphism (RRFLP): A novel technique for genotyping infectious laryngotracheitis virus (ILTV) live attenuated vaccines



Reverse restriction fragment length polymorphism (RRFLP): A novel technique for genotyping infectious laryngotracheitis virus (ILTV) live attenuated vaccines



Journal of Virological Methods 160(1-2): 119-124



A novel technique, the reverse restriction fragment length polymorphism (RRFLP) assay, was developed as a means of detecting specific informative polymorphic sites in the infectious laryngotracheitis virus (ILTV) genome. During the RRFLP procedure, DNA is digested with restriction enzymes targeting an informative polymorphic site and then used as template in a real-time polymerase chain reaction (PCR) with primers flanking the informative region. The analysis of the DeltaC(t) values obtained from digested and undigested template DNA provides the genotype of the DNA. In this study, the RRFLP assay was applied as a method to differentiate between the two types of infectious laryngotracheitis virus attenuated live vaccines. Sequence analysis of ILTV vaccines revealed an informative polymorphic site in the 5'-non-coding region of the infected cell protein (ICP4) gene. Unique AvaI and AlwI restriction enzyme sites were identified in the tissue culture origin and chicken embryo origin attenuated vaccines, respectively. These two informative polymorphic sites were used in a RRFLP assay to genotype rapidly and reproducibly ILTV attenuated live vaccines.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 055556425

Download citation: RISBibTeXText

PMID: 19433109

DOI: 10.1016/j.jviromet.2009.04.036


Related references

Efficacy of Live Virus Vaccines against Infectious Laryngotracheitis Assessed by Polymerase Chain Reaction-Restriction Fragment Length Polymorphism. Avian Diseases 47(2): 261-271, 2003

Efficacy of live virus vaccines against infectious laryngotracheitis assessed by polymerase chain reaction-restriction fragment length polymorphism. Avian Diseases 47(2): 261-271, 2003

Characterization of Infectious Laryngotracheitis Virus (Iltv) Isolates from Commercial Poultry by Polymerase Chain Reaction and Restriction Fragment Length Polymorphism (Pcr-Rflp). Avian Diseases 52(1): 59-63, 2008

Characterization of infectious laryngotracheitis virus (ILTV) isolates from commercial poultry by polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP). Avian Diseases 52(1): 59-63, 2008

Replication and transmission of live attenuated infectious laryngotracheitis virus (ILTV) vaccines. Avian Diseases 51(4): 905-911, 2008

Genome sequence comparison of two United States live attenuated vaccines of infectious laryngotracheitis virus (ILTV). Virus Genes 44(3): 470-474, 2012

Genotyping of infectious bronchitis virus isolates using reverse transcription-polymerase chain reaction and restriction fragment length polymorphism. Chinese Journal of Veterinary Science 19(6): 535-538, 1999

Analysis of Korean strains of infectious laryngotracheitis virus by nucleotide sequences and restriction fragment length polymorphism. Veterinary microbiology 83(4): 321-331, 2001

Epidemic of infectious laryngotracheitis in Italy: characterization of virus isolates by PCR-restriction fragment length polymorphism and sequence analysis. Avian Diseases 54(4): 1172-1177, 2010

Characterization of field strains of infectious laryngotracheitis virus in China by restriction fragment length polymorphism and sequence analysis. Journal of Veterinary Diagnostic Investigation 28(1): 46-49, 2016

Epidemic of Infectious Laryngotracheitis in Italy: Characterization of Virus Isolates by Pcr-Restriction Fragment Length Polymorphism and Sequence Analysis. Avian Diseases 54(4): 1172-1177, 2010

Characterization of infectious laryngotracheitis virus isolates from commercial poultry by polymerase chain reaction and restriction fragment length polymorphism. Avian Diseases: 1, 59-63, 2008

Rapid differentiation of vaccine strains and field isolates of infectious laryngotracheitis virus by restriction fragment length polymorphism of PCR products. Journal of Virological Methods 66(2): 179-186, 1997

Characterization of infectious laryngotracheitis virus isolates from the US by polymerase chain reaction and restriction fragment length polymorphism of multiple genome regions. Avian Pathology 36(2): 167-176, 2007

Restriction fragment length polymorphism analysis of multiple genome regions of Korean isolates of infectious laryngotracheitis virus collected from chickens. Poultry Science 92(8): 2053-2058, 2013