Infectious bronchitis virus Mass-type (GI-1) and QX-like (GI-19) genotyping and vaccine differentiation using SYBR green RT-qPCR paired with melting curve analysis

Andrea Laconi; Alinda J Berends; Esther C H de Laat; Tara A P M P Urselmann; Hélène M Verheije

doi:10.1016/j.jviromet.2019.113771

Infectious bronchitis virus Mass-type (GI-1) and QX-like (GI-19) genotyping and vaccine differentiation using SYBR green RT-qPCR paired with melting curve analysis

Andrea Laconi^*, Alinda J Berends, Esther C H de Laat, Tara A P M P Urselmann, Hélène M Verheije

^*Corresponding author for this work

Utrecht University

Research output: Contribution to journal › Article › Academic › peer-review

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.

Original language	English
Article number	113771
Pages (from-to)	1-7
Number of pages	7
Journal	Journal of Virological Methods
Volume	275
DOIs	https://doi.org/10.1016/j.jviromet.2019.113771
Publication status	Published - Jan 2020

Keywords

infectious bronchitis virus
genotyping
vaccine differentiation
RT-qPCR
melting curve

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/j.jviromet.2019.113771

Infectious bronchitis virusFinal published version, 1.77 MBLicence: Taverne

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title = "Infectious bronchitis virus Mass-type (GI-1) and QX-like (GI-19) genotyping and vaccine differentiation using SYBR green RT-qPCR paired with melting curve analysis",

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.",

keywords = "infectious bronchitis virus, genotyping, vaccine differentiation, RT-qPCR, melting curve",

author = "Andrea Laconi and Berends, {Alinda J} and {de Laat}, {Esther C H} and Urselmann, {Tara A P M P} and Verheije, {H{\'e}l{\`e}ne M}",

year = "2020",

month = jan,

doi = "10.1016/j.jviromet.2019.113771",

language = "English",

volume = "275",

pages = "1--7",

journal = "Journal of Virological Methods",

issn = "0166-0934",

publisher = "Elsevier BV",

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Infectious bronchitis virus Mass-type (GI-1) and QX-like (GI-19) genotyping and vaccine differentiation using SYBR green RT-qPCR paired with melting curve analysis. / Laconi, Andrea; Berends, Alinda J; de Laat, Esther C H et al.
In: Journal of Virological Methods, Vol. 275, 113771, 01.2020, p. 1-7.

Research output: Contribution to journal › Article › Academic › peer-review

TY - JOUR

T1 - Infectious bronchitis virus Mass-type (GI-1) and QX-like (GI-19) genotyping and vaccine differentiation using SYBR green RT-qPCR paired with melting curve analysis

AU - Laconi, Andrea

AU - Berends, Alinda J

AU - de Laat, Esther C H

AU - Urselmann, Tara A P M P

AU - Verheije, Hélène M

PY - 2020/1

Y1 - 2020/1

N2 - 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.

AB - 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.

KW - infectious bronchitis virus

KW - genotyping

KW - vaccine differentiation

KW - RT-qPCR

KW - melting curve

U2 - 10.1016/j.jviromet.2019.113771

DO - 10.1016/j.jviromet.2019.113771

M3 - Article

C2 - 31712091

SN - 0166-0934

VL - 275

SP - 1

EP - 7

JO - Journal of Virological Methods

JF - Journal of Virological Methods

M1 - 113771

ER -

Infectious bronchitis virus Mass-type (GI-1) and QX-like (GI-19) genotyping and vaccine differentiation using SYBR green RT-qPCR paired with melting curve analysis

Abstract

Keywords

UN SDGs

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