A combined cell‐culture and RT–PCR method for rapid detection of piscine nodaviruses

A combined cell‐culture and RT–PCR method for rapid detection of piscine nodaviruses A rapid method to detect piscine nodaviruses, the causal agents of viral nervous necrosis (VNN), at low virus levels is described. The reverse transcription–polymerase chain reaction (RT–PCR) technique required 104–105 TCID50 for detection of four strains representing different genotypes of piscine nodavirus: striped jack nervous necrosis virus (SJNNV), redspotted grouper nervous necrosis virus (RGNNV), tiger puffer nervous necrosis virus (TPNNV) and barfin flounder nervous necrosis virus (BFNNV). All the genotypic variants were isolated at the lowest titre (100 TCID50) by the E‐11 cell line, a clone derived from the SSN‐1 cell line, but it took approximately 10 days until the cytopathic effect (CPE) appeared in cells inoculated at low virus levels. When the virus was inoculated at various concentrations in E‐11 cells and incubated at 20 or 25 °C, cells inoculated previously with virus at a dose of 100 TCID50 became positive within 48 h incubation in the RT–PCR test. The present combined procedure of cell‐culture and RT–PCR techniques will be useful as a rapid and convenient method to detect infective viral particles from asymptomatic carriers or samples with low virus levels. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Journal of Fish Diseases Wiley

A combined cell‐culture and RT–PCR method for rapid detection of piscine nodaviruses

Journal of Fish Diseases, Volume 24 (4) – May 18, 2001

Loading next page...
 
/lp/wiley/a-combined-cell-culture-and-rt-pcr-method-for-rapid-detection-of-ioYsVxQbx8
Publisher
Wiley
Copyright
Copyright © 2001 Wiley Subscription Services, Inc., A Wiley Company
ISSN
0140-7775
eISSN
1365-2761
DOI
10.1046/j.1365-2761.2001.00291.x
Publisher site
See Article on Publisher Site

Abstract

A rapid method to detect piscine nodaviruses, the causal agents of viral nervous necrosis (VNN), at low virus levels is described. The reverse transcription–polymerase chain reaction (RT–PCR) technique required 104–105 TCID50 for detection of four strains representing different genotypes of piscine nodavirus: striped jack nervous necrosis virus (SJNNV), redspotted grouper nervous necrosis virus (RGNNV), tiger puffer nervous necrosis virus (TPNNV) and barfin flounder nervous necrosis virus (BFNNV). All the genotypic variants were isolated at the lowest titre (100 TCID50) by the E‐11 cell line, a clone derived from the SSN‐1 cell line, but it took approximately 10 days until the cytopathic effect (CPE) appeared in cells inoculated at low virus levels. When the virus was inoculated at various concentrations in E‐11 cells and incubated at 20 or 25 °C, cells inoculated previously with virus at a dose of 100 TCID50 became positive within 48 h incubation in the RT–PCR test. The present combined procedure of cell‐culture and RT–PCR techniques will be useful as a rapid and convenient method to detect infective viral particles from asymptomatic carriers or samples with low virus levels.

Journal

Journal of Fish DiseasesWiley

Published: May 18, 2001

References

You’re reading a free preview. Subscribe to read the entire article.


DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create folders to
organize your research

Export folders, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off