A Recombination Event, Induced In Ovo, Between a Low Passage Infectious Bronchitis Virus Field Isolate and a Highly Embryo Adapted Vaccine Strain

The possibility of genomic recombination among different strains of infectious bronchitis virus (IBV) was examined in ovo by coinfecting specific pathogen free embryonating chicken eggs with commonly used, embryo-adapted vaccine strains of IBV (Arkansas, Massachusetts, and Connecticut), and a Delawa...

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Bibliographic Details
Published in:Avian diseases 2003-10, Vol.47 (4), p.1282-1290
Main Authors: Estevez, C, Villegas, P, El-Attrache, J
Format: Article
Language:English
Subjects:
Infectious bronchitis virus
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Summary:The possibility of genomic recombination among different strains of infectious bronchitis virus (IBV) was examined in ovo by coinfecting specific pathogen free embryonating chicken eggs with commonly used, embryo-adapted vaccine strains of IBV (Arkansas, Massachusetts, and Connecticut), and a Delaware-072-like field virus isolated from a layer farm in Minnesota. Recombination was observed between the Massachusetts and the Delaware-072-like strains of the virus. The recombination event was assessed by reverse transcriptase-polymerase chain reaction (RT-PCR) using a combination of specific primers designed to flank a known recombination hot spot of the viral genomic sequence that codes for the S1 subunit of the spike envelope protein. The use of these primers allowed the detection of viruses that have undergone recombination around this hot spot. Cloning and sequencing of the RT-PCR product obtained was performed to confirm these results.Original Abstract: RESUMEN. Un evento de recombinacion, inducido in ovo, entre una cepa de campo del virus de bronquitis infecciosa con un numero bajo de pasajes y una cepa vacunal altamente adaptada a embriones. capital sigma e examino la posibilidad de recombinacion genomica entre diferentes cepas del virus de bronquitis infecciosa aviar in ovo mediante la inoculacion simultanea de embriones de pollos libres de patogenos especificos con una de varias cepas vacunales comunmente usadas en el campo (Arkansas, Massachussets y Connecticut), y una cepa de campo parecida a la cepa Delaware-072, obtenida a partir de una parvada de gallinas ponedoras en Minnesota. Se observo recombinacion entre la cepa vacunal Massachussets y la cepa de campo descrita. El evento de recombinacion fue demostrado mediante la prueba de reaccion en cadena por la polimerasa acoplada a transcripcion reversa usando iniciadores de reaccion disenados para amplificar la region del gen que codifica por la proteina S1 del virus que ha sido implicado en eventos de recombinacion en estudios previos. El uso de estos iniciadores de reaccion permitio la deteccion de los virus que presentaron recombinacion en esta zona del genoma. Se clonaron y se secuenciaron los productos amplificados mediante la reaccion en cadena por la polimerasa con el fin de confirmar los resultados obtenidos. double prime bbreviations:EID50 = embryo infectious dose 50% HPLC = high-performance liquid chromatography; IBV = infectious bronchitis virus; PBS = phosphate-buffered saline; RBC = red
ISSN:0005-2086
DOI:10.1043/0005-2086(2003)047(1282:AREIIO)2.0.CO;2