A consensus real-time RT-PCR for detection of all genotypic variants of yellow head virus of penaeid shrimp

被引：5
作者：
Wijegoonawardane, Priyanjalie K. M. [1 ]
Cowley, Jeff A. [1 ,2 ]
Walker, Peter J. [1 ,2 ]
机构：
[1] CSIRO, St Lucia, Qld 4067, Australia
[2] CSIRO, Australian Anim Hlth Lab, Geelong, Vic 3220, Australia
来源：
JOURNAL OF VIROLOGICAL METHODS | 2010年 / 167卷 / 01期
关键词：
Yellow head virus (YHV); Gill-associated virus (GAV); Quantitative real-time RT-PCR; Genotypic variation; Penaeus monodon; Genotypes; GILL-ASSOCIATED-VIRUS; TAURA-SYNDROME VIRUS; MONODON PRAWNS; GENOME SEQUENCE; NIDOVIRUS; DIFFERENTIATION; AUSTRALIA; THAILAND; GAV;
D O I：
10.1016/j.jviromet.2010.02.024
中图分类号：
Q5 [生物化学];
学科分类号：
071010 ; 081704 ;
摘要：
A real-time quantitative (q)RT-PCR employing consensus degenerate PCR primers was developed to detect all six genotypes known currently to comprise the yellow head virus (YHV) complex and found commonly in Penaeus monodon shrimp. The test primers possess only limited (eight-fold) degeneracy and target ORF1b gene sequences identified to be highly conserved amongst 57 strains of the six genotypes detected in P. monodon sourced from various regions of the Indo-Pacific. The qRT-PCR amplifies a 147 bp sequence and analysis of dilutions of synthetic genotype 2 RNA showed it to be 99.8% efficient and capable of detecting as few as 2.5 RNA copies reliably. As the test detects all six YH-complex genotypes, is extremely sensitive, capable of quantifying infection loads, and amenable to high-throughput application, it should prove useful for managing infections in P. monodon broodstock and seedstock used for aquaculture. Crown Copyright (C) 2010 Published by Elsevier B.V. All rights reserved.
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页码：5 / 9
页数：5
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