Quantitative real time PCR for the measurement of white spot syndrome virus in shrimp

S. V. Durand, Donald V Lightner

Research output: Contribution to journalArticle

101 Citations (Scopus)

Abstract

Quantitative real time PCR, recently developed in molecular biology, is applied in this paper to quantify the white spot syndrome virus (WSSV) in infected shrimp tissue. The WSSV content in moribund shrimp of all species tested (Penaeus stylirostris, P. monodon, P. vannamei) ranged from 2.0 × 104 to 9.0 × 1010 WSSV copies μg-1 of total DNA (n=26). In whole moribund post-larvae, 4.3 × 109 WSSV copies μg-1 of DNAwere detected which is equivalent to 5.7 × 1010 WSSV copies g-1 of post-larvae. The comparison of WSSV content between different tissues showed that muscle and hepatopancreas tissues contained 10 times less virus than gills, pleopods and haemolymph. With inocula of known virus content, bioassays by immersion challenge showed that a minimum of five logs of WSSV copies was necessary to establish disease in the challenged shrimp. In contrast, five logs of WSSV copies injected into shrimp muscle produced a LT-50 of 52 h. This real time polymerase chain reaction (PCR) technique is sensitive (four copies), specific (negative with DNA from shrimp baculoviruses and parvoviruses), dynamic (seven logs) and easy to perform (96 tests in < 4 h).

Original languageEnglish (US)
Pages (from-to)381-389
Number of pages9
JournalJournal of Fish Diseases
Volume25
Issue number7
DOIs
StatePublished - 2002

Fingerprint

White spot syndrome virus 1
White spot syndrome virus
polymerase chain reaction
Real-Time Polymerase Chain Reaction
virus
quantitative polymerase chain reaction
shrimp
Larva
Parvoviridae
Litopenaeus stylirostris
Penaeidae
Hepatopancreas
Viruses
muscle
Muscles
viruses
Parvovirus
Hemolymph
Penaeus monodon
Baculoviridae

Keywords

  • Penaeid shrimp
  • Quantitative real time PCR
  • WSSV

ASJC Scopus subject areas

  • Aquatic Science
  • veterinary(all)

Cite this

Quantitative real time PCR for the measurement of white spot syndrome virus in shrimp. / Durand, S. V.; Lightner, Donald V.

In: Journal of Fish Diseases, Vol. 25, No. 7, 2002, p. 381-389.

Research output: Contribution to journalArticle

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