PMID- 15158586
OWN - NLM
STAT- MEDLINE
DCOM- 20040907
LR  - 20061115
IS  - 0166-0934 (Print)
IS  - 0166-0934 (Linking)
VI  - 119
IP  - 2
DP  - 2004 Aug
TI  - Diagnosis of Ostreid herpesvirus 1 in fixed paraffin-embedded archival samples 
      using PCR and in situ hybridisation.
PG  - 65-72
AB  - In 1994, some of the high mortality episodes that affected oysters cultured in 
      France were associated with herpesviral infections. Through histology analysis, 
      however, viral presence could only be suspected and confirmation of histological 
      diagnosis by transmission electron microscopy was performed in only a few cases. 
      Subsequently, the characterisation and genome sequencing of Ostreid herpesvirus 1 
      (OsHV-1) made possible the development of specific molecular detection (PCR and 
      in situ hybridisation (ISH)). Using both molecular tools, attempts were made to 
      screen for OsHV-1 a number of fixed, paraffin-embedded oyster samples collected 
      and processed in 1994. The aim was to compare these techniques and to estimate 
      the accuracy of histology-based indication of viral infection. Existing DNA 
      extraction protocols were adapted for oyster samples and two pairs of specific 
      primers targeting small fragments (less than 200bp) were designed (C(9)/C(10) and 
      B(4)/B(3)). The poor consistency observed between the results of PCR with both 
      primer pairs was confirmed by statistical analysis. C(9)/C(10), which targets a 
      repeated region of the OsHV-1 genome, appears to be the primer of choice for 
      viral detection in archival samples. In situ hybridisation may furnish 
      complementary information concerning the localisation of viral foci. Under 
      certain conditions, retrospective examination of archival samples by molecular 
      techniques may therefore provide valuable epidemiological data.
FAU - Barbosa-Solomieu, V
AU  - Barbosa-Solomieu V
AD  - Institut Francais de Recherche pour l'Exploitation de la Mer (IFREMER), 
      Laboratoire de Genetique et Pathologie (LGP), 17390 La Tremblade, France.
FAU - Miossec, L
AU  - Miossec L
FAU - Vazquez-Juarez, R
AU  - Vazquez-Juarez R
FAU - Ascencio-Valle, F
AU  - Ascencio-Valle F
FAU - Renault, T
AU  - Renault T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Netherlands
TA  - J Virol Methods
JT  - Journal of virological methods
JID - 8005839
RN  - 0 (DNA, Viral)
RN  - 0 (Viral Proteins)
SB  - IM
MH  - Animals
MH  - DNA, Viral/*analysis
MH  - Herpesviridae/genetics/*isolation & purification
MH  - Herpesviridae Infections/diagnosis/*veterinary/virology
MH  - In Situ Hybridization
MH  - Ostreidae/*virology
MH  - Paraffin Embedding
MH  - Polymerase Chain Reaction/*methods
MH  - Viral Proteins/analysis
EDAT- 2004/05/26 05:00
MHDA- 2004/09/08 05:00
CRDT- 2004/05/26 05:00
PHST- 2003/08/27 00:00 [received]
PHST- 2004/02/11 00:00 [revised]
PHST- 2004/02/11 00:00 [accepted]
PHST- 2004/05/26 05:00 [pubmed]
PHST- 2004/09/08 05:00 [medline]
PHST- 2004/05/26 05:00 [entrez]
AID - S016609340400045X [pii]
AID - 10.1016/j.jviromet.2004.02.007 [doi]
PST - ppublish
SO  - J Virol Methods. 2004 Aug;119(2):65-72. doi: 10.1016/j.jviromet.2004.02.007.