A RANDOM GRID BASED MOLECULAR EPIDEMIOLOGICAL STUDY ON EBLV ISOLATES FROM GERMANY

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A RANDOM GRID BASED MOLECULAR EPIDEMIOLOGICAL
STUDY ON EBLV ISOLATES FROM GERMANY

• C. Freuling1, N. Johnson2, D. Marston2, T.
  Selhorst 1, L. Geue1, A. Fooks2, N.Tordo3 and T.
  Müller 1
• Friedrich-Loeffler-Institut, Federal Research
  Institute for Animal Health, D-16868
  Wusterhausen, Germany
• Veterinary Laboratories Agency (Weybridge),
  Surrey, United Kingdom
• Unité de la Rage and Laboratoire des Lyssavirus,
  Institut Pasteur, Paris, France

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Objective

• Molecular characterization of a representive
  panel of German EBLV isolates
• Germany has one of the highest numbers
• Geographic distribution as a key determination
  for selection in contrast to random sampling
• Spatiotemporal correlation to phylogenetic
  information

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Bat rabies in Europe 1977-2006
N 831
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Materials and Methods

• Application of a random grid with 30 km length
• Selection of 1 isolate per grid cell
• 48 out of 120 isolates selected
• Phylogenetic analysis based on complete N-gene
  sequences

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Results

• Identification of EBLV1b isolates from the
  southwest of Germany

5776N.seq
EBLV1a
EBLV1b
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Results
5776N.seq
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Results
5776N.seq
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Results
5776N.seq
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Results
5776N.seq
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Results
5776N.seq
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Results
5776N.seq
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Results

• short genomic insertions in the 3 UTR of the N
  gene
• 6 bp insertion in EBLV-1b (Johnson et al, 2007)
• 1 bp insertion in EBLV-1a

M
N
G
L
P
3
5
Is. No 976 AATTGGAAAGAAAAA------AA-CTAACAC
CACT Is. No 5006 AATTGAAAAGAAAAAGAAAAAAA-CT
AACACCACT Is. No 15571 AATTGAAAAGAAAAAGAAAAAA
A-CTAACACCACT Is. No 5776
AATTGGAAAGAAAAA------AAACTAACACCACT Is. No 3132
AATTGGAAAGAAAAA------AAACTAACACCACT
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Conclusions/Discussion

• Further confirmation of EBLV1b in southern
  Germany
• High sequence homology in N-Gene
• Indication for geographical clustering of EBLV1a
• No species differences
• Identification of short genomic insertions in the
  3 UTR of the N-gene
• Further research nessesary
• Other gene segments (e.g. G, ? pseudogene)
• More isolates from different time periods

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Thanks for attention!!