Publication:
Target for standard Thai PCR assay identical in 12 white spot syndrome virus (WSSV) types that differ in DNA multiple repeat length

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Issued Date

2005

Resource Type

Article

File Type

application/pdf

ISSN

1660934

DOI

10.1016/j.jviromet.2005.06.006

Other identifier(s)

2-s2.0-27744481024

Rights Holder(s)

Scopus

Bibliographic Citation

Journal of Virological Methods. Vol 130, (2005), p.79-82

Suggested Citation

Kiatpathomchai W., Taweetungtragoon A., Jittivadhana K., Wongteerasupaya C., Boonsaeng V., Flegel T.W. Target for standard Thai PCR assay identical in 12 white spot syndrome virus (WSSV) types that differ in DNA multiple repeat length. Journal of Virological Methods. Vol 130, (2005), p.79-82. doi:10.1016/j.jviromet.2005.06.006 Retrieved from: https://hdl.handle.net/20.500.14740/6039

Title

Target for standard Thai PCR assay identical in 12 white spot syndrome virus (WSSV) types that differ in DNA multiple repeat length

Author(s)

Kiatpathomchai W.
 Taweetungtragoon A.
 Jittivadhana K.
 Wongteerasupaya C.
 Boonsaeng V.
 Flegel T.W.

Abstract

A Thai PCR detection method (WSSV-232) yielding a 232 bp amplicon has been used for detection of white spot syndrome virus (WSSV) since 1996. It targets ORF 91 in the full sequence of the only Thai WSSV isolate at GenBank (AF369029). At the beginning of 2002, some Thai shrimp farmers complained that ponds stocked with WSSV-232 PCR negative post-larvae (PL) later suffered WSSV disease outbreaks. Although these outbreaks may have resulted from horizontal transmission of WSSV after stocking, it was also possible that they resulted from false negative PCR test results due to genetic changes at the PCR-assay target after the first appearance of WSSV in Thailand in 1995. Indeed, recent results have revealed at least 12 WSSV variants in Thailand that can be distinguished based on differences in DNA multiple repeat lengths in ORF 94 (GenBank AF369029). To test for variation in the WSSV-232 target sequence in ORF 91, 20 DNA extracts derived from field samples and representing 9 of the WSSV DNA multiple repeat groups were subjected to PCR amplification and sequencing using primers that generated a 403 bp amplicon covering the target for the WSSV-232 assay. An additional three repeat types were included from archived material. Analysis revealed that the 232 bp target sequence in ORF 91 was unchanged in all of the 12 types tested and that the original WSSV-232 detection system was still valid. Thus, any false negative PCR test results leading to farmer complaints would probably have arisen from small sample sizes and low sensitivity of the single-step PCR assay. If so, false negative results could be reduced by the use of nested PCR assays with larger PL sample sizes. © 2005 Elsevier B.V. All rights reserved.

Subject(s)

Virus DNA
 Agricultural worker
 Article
 Controlled study
 Gene sequence
 Genetic variability
 Laboratory diagnosis
 Nonhuman
 Nucleotide repeat
 Nucleotide sequence
 Open reading frame
 Polymerase chain reaction
 Priority journal
 Sample size
 Sensitivity analysis
 Validation process
 Virus
 Virus detection
 Virus gene
 Virus strain
 White spot syndrome virus
 Animals
 Base Sequence
 DNA, Viral
 Molecular Sequence Data
 Open Reading Frames
 Penaeidae
 Polymerase Chain Reaction
 Sequence Analysis, DNA
 Thailand
 White spot syndrome virus 1
 Decapoda (Crustacea)
 DNA viruses
 Shrimp white spot syndrome virus

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URI

https://hdl.handle.net/20.500.14740/6039

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Scopus: Year 1983-2021

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