Laboratório de Biologia Molecular Aplicada - LAPLIC, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil; Programa de Pós-Graduação em Bioquímica, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.
Laboratório de Biologia Molecular Aplicada - LAPLIC, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil; Escola Agrícola de Jundiaí, Universidade Federal do Rio Grande do Norte, Macaíba, RN, Brazil.
J Invertebr Pathol. 2019 Mar;162:55-63. doi: 10.1016/j.jip.2019.02.003. Epub 2019 Feb 6.
White spot syndrome virus (WSSV) is one of the major challenges faced by global shrimp farming in recent decades. The characterization of WSSV genetic variability has been used to determine viral dispersion and is a promising method to determine the association between genotype and virulence. The major variable regions that have been used as markers to differentiate the WSSV genomes include the VNTR loci inside ORF94, ORF75, ORF125, and insertions/deletions interspersing ORF14/15 and ORF23/24. The primers used to amplify these regions were described at least 10 years ago, but some of them do not work efficiently to identify new WSSV variants. The objective of this work was to develop improved PCR primers for WSSV genotyping based on sequence alignments that include new sequences described in recent years. We validated these new primers in a pilot study to verify the genetic variability of the WSSV in Rio Grande do Norte state (northeast Brazil), and efficiency was compared to that of other previously described primers. We confirmed that the primers we developed were more efficient for genotype Brazilian WSSV isolates, enabling us to genotype a larger number of samples. In addition, our results also introduce new data about the genetic characterization of the WSSV isolates that occur in the northeastern region of Brazil.
白斑综合征病毒（WSSV）是近几十年来全球对虾养殖业面临的主要挑战之一。WSSV 遗传变异的特征已被用于确定病毒的传播，并且是确定基因型与毒力之间关联的有前途的方法。已被用作区分 WSSV 基因组的标记的主要可变区包括 ORF94、ORF75、ORF125 内的 VNTR 基因座，以及插入/缺失散布在 ORF14/15 和 ORF23/24 之间。用于扩增这些区域的引物至少在 10 年前就已描述，但其中一些引物不能有效地识别新的 WSSV 变体。这项工作的目的是基于包括近年来描述的新序列的序列比对，开发用于 WSSV 基因分型的改进 PCR 引物。我们在一项试点研究中验证了这些新引物，以验证巴西北部里奥格兰德州（巴西东北部）的 WSSV 的遗传变异性，并将其与其他先前描述的引物的效率进行了比较。我们证实，我们开发的引物对巴西 WSSV 分离株的基因型更有效，使我们能够对更多的样本进行基因分型。此外，我们的结果还介绍了有关发生在巴西东北部地区的 WSSV 分离株遗传特征的新数据。
