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.
BACKGROUND
A 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.


METHODS
In 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.


RESULTS
We 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.


CONCLUSIONS
The 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.
Unit of Avian Pathology, Agronomic and Veterinary Institute Hassan II, B.P. 6202, Rabat, Morocco.|Laboratory of Virology, Microbiology and Quality/Ecotoxicology & Biodiversity, Faculty of Sciences and Techniques-University Hassan II Mohammedia, PO Box 146, Quartier Yasmina-Mohammedia, 20650, Casablanca, Morocco.
Siham
S
Fellahi
Laboratory of Molecular Biology-Society of Biological Products and Veterinary Pharmaceuticals (Biopharma), B.P. 4569, Km 2, Route de Casa, Rabat, Morocco.
Mehdi
M
El Harrak
Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, B-8200 Research Plaza, Fort Detrick, Frederick, MD, 21702, USA.
Jens H
JH
Kuhn
Laboratory of Molecular Biology-Society of Biological Products and Veterinary Pharmaceuticals (Biopharma), B.P. 4569, Km 2, Route de Casa, Rabat, Morocco.
Ghizlane
G
Sebbar
Laboratory of Epidemiology and Clinical Research. Faculty of Medicine, B.P. 1014, 4, Avenue Ibn Battouta, Rabat, Morocco.
El Arbi
el A
Bouaiti
Laboratory of Virology, Microbiology and Quality/Ecotoxicology & Biodiversity, Faculty of Sciences and Techniques-University Hassan II Mohammedia, PO Box 146, Quartier Yasmina-Mohammedia, 20650, Casablanca, Morocco.
Khadija
K
Khataby
Unit of Avian Pathology, Agronomic and Veterinary Institute Hassan II, B.P. 6202, Rabat, Morocco.
Ouafae Fassi
OF
Fihri
Unit of Avian Pathology, Agronomic and Veterinary Institute Hassan II, B.P. 6202, Rabat, Morocco.
Mohammed
M
El Houadfi
Laboratory of Virology, Microbiology and Quality/Ecotoxicology & Biodiversity, Faculty of Sciences and Techniques-University Hassan II Mohammedia, PO Box 146, Quartier Yasmina-Mohammedia, 20650, Casablanca, Morocco. m.ennaji@yahoo.fr.
My Mustapha
MM
Ennaji
D000818:Animals; D052160:Benzothiazoles; D001715:Bird Diseases / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology*; D002645:Chickens; D018352:Coronavirus Infections / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology*; D000086462:Coronavirus Nucleocapsid Proteins; D003959:Diamines; D004196:Disease Outbreaks / Q000517:prevention & control; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D001351:Infectious bronchitis virus / Q000235:genetics*; D009018:Morocco / Q000453:epidemiology; D019590:Nucleocapsid Proteins / Q000235:genetics; D009930:Organic Chemicals; D011804:Quinolines; D012367:RNA, Viral / Q000235:genetics; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D052160:Benzothiazoles; D000086462:Coronavirus Nucleocapsid Proteins; D003959:Diamines; D019590:Nucleocapsid Proteins; D009930:Organic Chemicals; D011804:Quinolines; D012367:RNA, Viral; C098022:SYBR Green I
Coronavirus; Gammacoronavirus; IBV; Infectious bronchitis virus; RT-PCR; Real time RT-PCR; SYBR green