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OJVRTM
Online Journal of
Veterinary Research©
Volume 21(10):644-656, 2017.
In
situ PCR for detection and differentiation of infectious bursal disease
virus strains in chickens
Elawad A.Hussein1,
Mohd Hair-Bejo1, 3*, Pit S. Liew1, Abdul
R. Omar 1, 3, Siti
S. Arshad1, Ideris Aini2,3
Department(s)
1Veterinary Pathology and Microbiology, 2Veterinary
Clinical Studies, Faculty of Veterinary Medicine, 3Institute of
Bioscience, Universiti Putra Malaysia, Selangor,
Malaysia *Corresponding author:
Department of Veterinary Pathology and Microbiology, Faculty of Veterinary
Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia. E-mail address: mdhair@upm.edu.my;
husseinabdallah25@gmail.com (M.
Hair-Bejo)
ABSTRACT
Hussein EA, Hair-Bejo M1, Liew PS, Omar AR, Arshad SS, Aini
I., In situ PCR for detection and differentiation of infectious bursal disease
virus strains in chickens, Onl J Vet Res., 21(10):644-656,
2017. An In situ PCR method for detection and
differentiation of infectious bursal disease virus (IBDV) strains is described.
In one study, 15 specific pathogen free (SPF) 14 day old chickens were infected
orally with very virulent (vv) IBDV strain with a
titre of 107.5 EID50/0.1 mL.
Six non-infected chickens were used as controls. Chickens were sacrificed at
various intervals and tissue samples taken for histological examination. Immunoperoxidase staining (IPS) was done, and an In situ PCR was developed using a
specific probe for IBDV’s VP1 gene. The In situ PCR was significantly (p < 0.05) more sensitive than IPS. In another
study to differentiate strains by In
situ PCR, ten, 42 day old SPF
chickens were infected with virulent (104.83 EID50/0.1
mL) or classical (ca) NDV strains (103.0 EID50/0.1 mL)
with 5 non-infected controls. Tissue samples infected with virulent, classic
and controls tested with a virulent specific probe were positive only in
tissues infected by the virulent strain whereas classical strain probe were
positive only to tissue infected by classical strain. These results suggest
that our In situ PCR differentiated
virulent from classical NDV strains.
Key words: In
situ PCR, specific probe, infectious bursal disease virus (IBDV), very
virulent IBDV, classical IBDV, detection and differentiation.
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