Comparison of SYBR green I real-time RT-PCR with conventional agarose gel-based RT-PCR for the diagnosis of infectious bronchitis virus infection in chickens in MoroccoReport as inadecuate




Comparison of SYBR green I real-time RT-PCR with conventional agarose gel-based RT-PCR for the diagnosis of infectious bronchitis virus infection in chickens in Morocco - Download this document for free, or read online. Document in PDF available to download.

BMC Research Notes

, 9:231

VeterinaryResearch

Abstract

BackgroundA rapid, sensitive, and specific molecular method for the diagnosis of infectious bronchitis virus IBV infection is important in curbing infectious bronchitis outbreaks in Morocco and other countries.

MethodsIn this study, an easy-to-perform SYBR green I real-time reverse transcriptase polymerase chain reaction RT-PCR targeting the nucleocapsid gene of IBV was developed and compared with conventional agarose gel-based RT-PCR for the detection of IBV infection.

ResultsWe found that the SYBR green I real-time RT-PCR was at least 10 times more sensitive than the agarose gel electrophoresis detection method. The assay exhibited high specificity for IBV infection. All negative controls, such as Newcastle disease virus, infectious bursal disease virus, and avian influenza virus, were not detected.

ConclusionThe SYBR green I real-time RT-PCR test described herein can be used to rapidly distinguish IBV from other respiratory pathogens, which is important for diagnosis and control of infectious bronchitis outbreaks in Morocco. The test is a valuable and useful method as a routine assay for diagnosis of clinical IBV infection in commercial chickens.

KeywordsCoronavirus Gammacoronavirus IBV Infectious bronchitis virus SYBR green Real time RT-PCR RT-PCR Abbreviationsbpbase pairs

Ctcycle threshold

ELISAenzyme-linked immunosorbent assay

FLUAVavian influenza virus

IBinfectious bronchitis

IBDVinfectious bursal disease virus

IBVinfectious bronchitis virus

Mmembrane protein

Nnucleocapsid protein

NCnegative control

NDVnewcastle disease virus

RNAribonucleic acid

RT-PCRreverse transcriptase-polymerase chain reaction

Sspike protein

Mohammed El Houadfi and My Mustapha Ennaji contributed equally to this work

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Author: Siham Fellahi - Mehdi El Harrak - Jens H. Kuhn - Ghizlane Sebbar - El Arbi Bouaiti - Khadija Khataby - Ouafae Fassi Fih

Source: https://link.springer.com/



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