Optimizing the nested PCR method for Decapod iridescent virus 1 (DIV1) targeting ATPase gene by reselecting the inner primers

IF 3.6 3区 生物学 Q1 ZOOLOGY
Jing-Yi Xing , An-Qi Li , Xiao-Meng Guo , Meng Wang , Xin Guan , Liang Qiu , Qing-Li Zhang , Jie Huang
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引用次数: 0

Abstract

DIV1 has the characteristics of fast transmission and a broad host range. Its infection leads to a high mortality rate, posing a serious threat to the global crustacean aquaculture industry. In order to increase the accuracy of DIV1 detection and reduce the difficulty of result interpretation, this study modified the original nested PCR method targeting the DIV1 ATPase gene. The internal primers for the nested PCR were redesigned to produce a 338 bp amplification product in the second step PCR, effectively distinguishing the target band from primer dimers. The newly established nested PCR method exhibits strong specificity and high sensitivity, with a detection limit as low as 1.37 × 101 copies/reaction. The developed nested PCR assay provides new technical support for the accurate detection of DIV1 in global crustacean aquaculture.

Abstract Image

通过重新选择内部引物,优化针对十足类虹彩病毒 1 (DIV1) ATPase 基因的嵌套 PCR 方法。
DIV1 具有传播速度快、宿主范围广的特点。其感染后死亡率极高,对全球甲壳类水产养殖业构成严重威胁。为了提高 DIV1 检测的准确性并降低结果解读的难度,本研究对原有的针对 DIV1 ATPase 基因的巢式 PCR 方法进行了改进。重新设计了巢式 PCR 的内部引物,使其在第二步 PCR 中产生 338 bp 的扩增产物,从而有效地将目标条带与引物二聚体区分开来。新建立的巢式 PCR 方法特异性强、灵敏度高,检测限低至 1.37 × 101 拷贝/反应。所开发的巢式 PCR 检测方法为在全球甲壳类水产养殖中准确检测 DIV1 提供了新的技术支持。
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来源期刊
CiteScore
6.10
自引率
5.90%
发文量
94
审稿时长
1 months
期刊介绍: The Journal of Invertebrate Pathology presents original research articles and notes on the induction and pathogenesis of diseases of invertebrates, including the suppression of diseases in beneficial species, and the use of diseases in controlling undesirable species. In addition, the journal publishes the results of physiological, morphological, genetic, immunological and ecological studies as related to the etiologic agents of diseases of invertebrates. The Journal of Invertebrate Pathology is the adopted journal of the Society for Invertebrate Pathology, and is available to SIP members at a special reduced price.
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