Year: 2018 | Month: October | Volume 8 | Issue 5

Restriction Fragment Length Polymorphism Analysis of Isolates of Infectious Bursal Disease Viruses from Southern Region of India


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Abstract:

The Restriction Fragment Length Polymorphism (RFLP) is used for the differentiation of classical virulent (cv), virulent (v) and very virulent (vv) strains of Infectious Bursal Disease Virus (IBDV) isolates collected from recent outbreaks in southern region of India. In the present study, five different isolates (BGE15, EDE14, RPM14, MDI14 and THI14) of IBDV strains were subjected for genotyping along with vaccine (Georgia, intermediate strain) by performing RT-PCR for amplification of a 743 bp from the hypervariable region of VP2 gene followed by restriction enzyme digestion with seven different enzymes (HhaI, SacI, SspI, StyI, BspMI, StuI and TaqI) for the differentiation of classical, virulent and very virulent strains of IBDV. The RT-PCR product obtained from all the five isolates were not cleaved by SspI and SacI enzyme and thus revealed the absence of restriction enzyme (RE) site for SspI and SacI enzyme, respectively. The HhaI enzyme cleaved vaccine and field isolates with similar restriction profiling pattern. The StuI enzyme did not digest vaccine strain whereas field isolates were cleaved by this enzyme. The BspM1 was not able to differentiate the field isolates from vaccine strain. TaqI enzyme cleaved both vaccine and field isolates of IBDV with different profile pattern. The StyI enzyme showed single RE site on vaccine strain and other field isolates with similar RE pattern. Thus, from the present study, it may be concluded that all the isolates belonged to vvIBDV and they do not have site for SspI marker.



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