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J Virol Methods ; 275: 113771, 2020 01.
Article in English | MEDLINE | ID: mdl-31712091

ABSTRACT

Infectious Bronchitis Virus (IBV) is a highly contagious virus of chicken, causing huge economic losses in the poultry industry. Many genotypes circulate in a given area, and optimal protection relies on vaccination with live attenuated vaccines of the same genotype. As these live vaccines are derived from field viruses and circulate, understanding the prevalence of different IBV genotypes in any area is complex. In a recent study, the genome comparison of an IBV QX vaccine and its progenitor field strain led to the identification of vaccine markers. Here we developed a simplex SYBRgreen RT-qPCR assay for differentiation between QX-like field and vaccine strains and a multiplex SYBRgreen RT-qPCR assay for IBV genotyping with melting curve analysis, as each virus produced distinct and reliable melting peaks. Both the simplex and the multiplex assays showed excellent efficiency, sensitivity and specificity representing a low cost diagnostic tool for IBV genotyping and vaccine differentiation.


Subject(s)
Coronavirus Infections/veterinary , Infectious bronchitis virus/genetics , Molecular Diagnostic Techniques/methods , Poultry Diseases/diagnosis , Real-Time Polymerase Chain Reaction/methods , Viral Vaccines/genetics , Animals , Chickens , Coronavirus Infections/diagnosis , Coronavirus Infections/virology , Genotype , Infectious bronchitis virus/isolation & purification , Molecular Diagnostic Techniques/veterinary , Poultry Diseases/virology , Real-Time Polymerase Chain Reaction/veterinary , Sensitivity and Specificity , Vaccines, Attenuated/genetics , Vaccines, Attenuated/isolation & purification , Viral Proteins/genetics , Viral Vaccines/isolation & purification
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