Yan Dongchun, Tang Kathy F J, Lightner Donald V
College of Life Science, Ludong University, Yantai 264025, PR China.
J Invertebr Pathol. 2009 Oct;102(2):97-100. doi: 10.1016/j.jip.2009.07.008. Epub 2009 Jul 19.
A real-time PCR method was developed to detect monodon baculovirus (MBV) in penaeid shrimp. A pair of MBV primers to amplify a 135bp DNA fragment and a TaqMan probe were developed. The primers and TaqMan probe were specific for MBV and did not cross react with Hepatopancreatic parvovirus (HPV), White spot syndrome virus (WSSV), Infectious hypodermal and haematopoietic virus (IHHNV) and specific pathogen free (SPF) shrimp DNA. A plasmid (pMBV) containing the target MBV sequence was constructed and used for determination of the sensitivity of the real-time PCR. This real-time PCR assay had a detection limit of one plasmid MBV DNA copy. Most significantly, this real-time PCR method can detect MBV positive samples from different geographic locations in the University of Arizona collection, including Thailand and Indonesia collected over a 13-year period.
开发了一种实时PCR方法来检测对虾中的斑节对虾杆状病毒（MBV）。设计了一对用于扩增135bp DNA片段的MBV引物和一个TaqMan探针。这些引物和TaqMan探针对MBV具有特异性，不会与肝胰腺细小病毒（HPV）、白斑综合征病毒（WSSV）、传染性皮下和造血组织坏死病毒（IHHNV）以及无特定病原体（SPF）的对虾DNA发生交叉反应。构建了一个包含目标MBV序列的质粒（pMBV），并用于测定实时PCR的灵敏度。这种实时PCR检测方法的检测限为一个质粒MBV DNA拷贝。最重要的是，这种实时PCR方法可以检测亚利桑那大学收集的来自不同地理位置的MBV阳性样本，包括在13年期间收集的来自泰国和印度尼西亚的样本。
