J Vet Sci.  2012 Dec;13(4):405-412.

Molecular characterization of two Bangladeshi infectious bursal disease virus isolates using the hypervariable sequence of VP2 as a genetic marker

• Islam MT
• Le TH
• Rahman MM
• Islam MA

Abstract

Two Bangladeshi infectious bursal disease virus (IBDV) isolates collected in 2007, termed GB1 and GB3, were subjected to comparative sequencing and phylogenetic analyses. Sequence analysis of a 474-bp hypervariable region in the VP2 gene revealed that among four major amino acid substitutions observed in the strains, two were unique to GB1 and GB3 (Ser217Leu and Ala270Thr) while one substitution was only found in GB1 (Asn299Ser). Among IBDVs from Bangladesh including GB1 and GB3, the rate of identity and homology was around 97~99%. The amino acid sequences of GB1 and GB3 differ from those of previous Bangladeshi IBDV isolates and contain amino acid substitutions Pro222Ala and Asn299Ser (in GB3 only). Phylogenetic analysis revealed that GB1 and GB3 are grouped with other very virulent IBDVs of European and American origin in contrast to two previously isolated Bangladeshi IBDV strains (GenBank accession Nos. AF362776 and AF260317), which belong to the Asian group. It was concluded that GB1 and GB3 belong to a very virulent group of IBDVs. However, amino acid sequences of GB1 and GB3 differ from those of the other Bangladeshi IBDVs by one or two amino acids encoded in the hypervariable region of the VP2 gene.

Keyword

chickens; infectious bursal disease virus; molecular characterization

MeSH Terms

Amino Acid Sequence
Amino Acid Substitution
Amino Acids
Asian Continental Ancestry Group
Bangladesh
Chickens
Genetic Markers
Humans
Infectious bursal disease virus
Sequence Analysis

Figure

• 

  Fig. 1 Alignment and comparison of predicted amino acid sequences for the hypervariable region of the VP2 gene of 17 IBDV isolates. Amino acid sequence of the isolate 52-70 (US) is shown on top. Identities of the subsequently aligned sequences are indicated by dots and differences are represented by single letters. Key amino acids examined in this study are indicated by numbers and arrows. The amino acid sequence (underlined in the top sequence) containing the neutralizing epitopes including major (A and B) and minor (1, 2, and 3) hydrophilic regions in the 17 isolates (boxed regions) is shown. US: USA, DE: Germany, BD: Bangladesh, VN: Vietnam, NP: Nepal, IN: India, HK: Hong Kong, KR: Korea, PK: Pakistan.

• 

  Fig. 2 Phylogenetic tree constructed with MEGA 4.1 illustrating the relationships among Bangladeshi IBDV isolates (GB1 and GB3) and 75 other strains of different geographical origins. Distance matrices were constructed using the Kimura's two-parameter model and trees were constructed using the neighbor-joining (NJ) algorithm based on the pairwise nucleotide differences in the VP2 variable region. Units at the bottom of the tree indicate the number of substitution events. The length of each pair of branches represents the distance between sequence pairs. The dataset was resampled 1,000 times using the bootstrap method. Accession numbers of the sequences and countries of origin are marked at end of strains when known.

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