SYBR Green-based duplex real time PCR method for detection of Infectious bronchitis virus, Mass serotype

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dc.contributor.authorRajalakshmi, S
dc.contributor.authorShilpa, P
dc.contributor.authorJohn Kirubaharan, J
dc.contributor.authorVidhya, M; et al.
dc.contributor.authorTANUVAS
dc.date.accessioned2020-03-05T10:32:52Z
dc.date.available2020-03-05T10:32:52Z
dc.date.issued2020-02
dc.descriptionTNV_20thMVC_PP_Feb-2020_OP51en_US
dc.description.abstractInfectious bronchitis (IB) is a highly contagious economically important viral disease of poultry, caused by Infectious bronchitis virus (IBV) belonging to the family Coronaviridae. The Massachusetts (Mass) serotype of IBV is being used worldwide for regular vaccination against IB since several years. The continuous application of the Mass strain of IBV for vaccination and mutating nature of virus has yielded a selection pressure, resulting in the emergence of field strains of non-Mass serotypes. Currently, rapid detection of IBV infection in the poultry flocks amidst other upper-respiratory avian pathogens is a major challenge, which demands sensitive and rapid diagnostic methods. The SYBR Green – based RT PCR assay has been proven to be one of the most effective and sensitive tools for such a diagnosis. A duplex SYBR Green – basedreal time RT PCR assay has been designed for the simultaneous detection of IBV and identification of the Mass serotype in a single reaction based on melt curve analysis. IBV-specific UTR primers and IBV-Mass- S1-specific primers used in the duplex real time RT-PCR yielded curves of amplification with two specific melting curves (Tm1 = 81oC ± 0.5oC and Tm2 = 84oC ± 0.7oC) for IBV – Mass serotype. Owing to the fact that IBV serotypes other than Mass do not exist in India, the assay was standardised using a known Mass serotype (H120 vaccine strain) and a well-characterised field isolate IBV –B17. The assay did not yield amplification curves when viruses other than IBV were assessed. This duplex realtime PCR can thus be used as a rapid molecular diagnostic tool for identification of IB outbreaks. Further evaluation of the assay for the simultaneous detection and differentiation of IBV Mass and non-Mass serotypes would throw more light for the effective control of IB, especially in vaccinated flocks.en_US
dc.identifier.urihttp://krishikosh.egranth.ac.in/handle/1/5810144284
dc.keywords20th Indian Veterinary Congress, Annual Conference of IAAVR, TANUVASen_US
dc.language.isoenen_US
dc.pages1en_US
dc.subjectVeterinary Scienceen_US
dc.titleSYBR Green-based duplex real time PCR method for detection of Infectious bronchitis virus, Mass serotypeen_US
dc.title.alternativeNational Symposium on Veterinary Research Priorities in Translational Animal Health, Production and Food Safetyen_US
dc.typePresentationen_US
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