Proficiency tests in molecular biology

Year

Technique

Host

Pathogen

2008

PCR

Ostrea edulis

Bonamia ostreae

2010

PCR-RFLP

Ostrea edulis

Marteilia refringens

2011

RT-PCR

Crassostrea gigas

Herpes virus OsHV-1

2013

PCR & RT-PCR

Ostrea edulis

Bonamia sp.

2015

RT-PCR

Crassostrea gigas

Vibrio aestuarianus

2017

PCR & RT-PCR

Ostrea edulis

Marteilia refringens &

Bonamia sp.

  • The 2008 proficiency test was devoted to the detection of  Bonamia ostreae by  PCR. The test included:
    1. the DNA extraction from gill tissue collected from 30 flat oysters, Ostrea edulis fixed in ethanol,
    2. the PCR test of these 30 samples using Cochennec et al. (2000) protocol.
  • The 2010 test was organised for the detection of Marteilia refringens by PCR-RFLP. The test included:
    1. the DNA extraction from tissue collected from 12 flat oysters Ostrea edulis and 12 mussels Mytilus edulis fixed in ethanol.
    2. the PCR test of these 24 samples using Le Roux et al. (2001) protocol see SOPs page.
  • The 2011 proficiency test was organised for the detection and quantification of herpes virus OsHV-1 in the Pacific oyster Crassostrea gigas by Real Time PCR. The test included:
    1. the DNA extraction from 24 suspensions of ethanol-fixed mantle tissue of oyster Crassostrea gigas.
    2. the PCR test of these 24 samples using Real Time PCR (see protocol on the SOPs page).
  • The 2013 test was organised for the detection of Bonamia sp.in the flat oyster Ostrea edulis by Conventional PCR (protocol available on the SOPs page). Optionally, the ILC aimed at comparing different PCR techniques for the detection of Bonamia sp. and the characterization of the species B. ostreae and B. exitiosa. The test included:
    1. the DNA extraction from 24 suspensions of ethanol-fixed gills tissues of oyster O. edulis.
    2. the PCR test of these 24 samples using Cochennec et al. (2000) protocol (see SOPs page).
    3. Optionnally:  the PCR test of these 24 samples using one or several other methods (a list was provided) and the characterisation at the species level (B. ostreae or B. exitiosa).
  • The 2015 proficiency test was organised for the detection Vibrio aestuarianus in the Pacific oyster Crassostrea gigas by Real Time PCR. The test included:
    1. the DNA extraction from 24 suspensions of ethanol-fixed tissue tissues (gills and mantles) of oyster Crassostrea gigas.
    2. the PCR test of these 24 samples using Real Time PCR (see protocol on the SOPs page).

In 2017, an Inter Laboratory Comparison (ILC) test is on-going to test the competency of participants regarding the detection of Marteilia refringens and Bonamia sp. in flat oyster (Ostrea edulis) tissues by PCR. Participant can choose the PCR assay they want to perform the test (see protocols on the SOPs page). Optionally, participants are invited to test a new multiplex real-time PCR recently developed by the EURL for mollusc diseases, allowing the simultaneous detection of Marteilia refringens and Bonamia sp. (primers and probes for this new PCR assay were provided).

The test consists of 24 suspensions of ethanol-fixed  tissues (gills and digestive glands) of oyster O. edulis , and two positive controls.

Reports are available in the "NRL's section > Proficiency test > Result and reports" page (access limited to the European NRLs by login and password).