PCR and virus detection
From: Olivier Mueller omueller@fastnet.ch
To: shrimp@yahoogroups.com
Date: 20 July 2004
QUESTION:
In the framework of our project of organic shrimp farming in Vietnam I am searching primers and positive controls to check by PCR a stock of P. monodon breeders. I am looking for primers to detect the most important virus that could infect our broodstock. Anybody can provide us information where to obtain these primers or to help us to get them? Thanks in advance for your help.
Olivier Mueller
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COMMENTS 1:
The Shrimp Biotechnology Business Unit (SBBU) in
Bangkok, Thailand, provides PCR kits in the form of primer sets or ready to
use kits for the major viruses encountered in shrimp farming. We also
provide sample analysis (PCR, histology, IHS...) in our laboratories and
different services linked to shrimp health management.
Please contact us directly offlist at ptacon@shrimpbiotec.com
You can also visit our website: www.shrimpbiotec.com
Philippe Tacon
e-mail: phil_sbbu@yahoo.fr
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COMMENTS 2:
See with DiagXotics www.diagxotics.com
and Farming Intelligene Tech www.iq2000kit.com
Thay have PCR and Dot-Blot.
Eric Pinon
e-mail: epinon@serviceaqua.com
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COMMENTS 3:
For more information on WSSV by PCR detection, also
check www.nicegarden.com.tw and http://www.enbiotec.co.jp
and www.magellan.com.tw
e-mail: zuridah@asian-agribiz.com
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COMMENTS 4:
On this interesting topic, does anyone have
information on the ability of PCR to detect WSV in postlarvae of different
ages? i.e., Is it not the same to run PCR on 5 day old vs 15 or 30 day old
PLs? I know conventional wisdom is the larger the PL the better relaibility
of the tests, but how much better in relation to age? Is there anything
published on this topic?, and what are the minimum PL ages for
"reasonable" reliability?
Lorenzo Juarez, SyAqua-Mexico
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COMMENTS 5:
Actually to run a reliable PCR, for WSSV for example,
it is not the
size, or age, of the PL that matters but how much nucleic acid you put
in your PCR vial. Basically you can start from nauplii if you want,
but you have to use a lot of them. For a similar amount of DNA, the
younger the PL, the more you have to use to extract DNA.
DNA extraction is also a very important step for the reliability of
the PCR reaction. Phenol-chloroform or membrane extraction should be
preferred than crude extraction (often leading to the inhibition of the
PCR reaction).
Philippe Tacon
e-mail: phil_sbbu@yahoo.fr