Diseases of Aquatic Organisms | |
Detection and surveillance of viral hemorrhagic septicemia virus using real-time RT-PCR. I. Initial comparison of four protocols | |
Emily R. Cornwell1  Robert Kim1  Kathy Toohey-Kurth1  Mohamed Faisal1  Andrew E. Goodwin1  Geoffrey H. Groocock1  Monica M. Reising1  Cem Giray1  Gwenn E. Merry1  01  Isaac Standish1  Nicholas B. D. Phelps1  Janet V. Warg1  Rodman G. Getchell1  Yan Zhang1  Travis Clement1  Angela Cruz1  | |
关键词: VHSV; Surveillance; Real-time RT-PCR; Analytical sensitivity; Analytical specificity; Validation; | |
DOI : 10.3354/dao02753 | |
学科分类:生物科学(综合) | |
来源: Inter-Research | |
【 摘 要 】
ABSTRACT: Eight laboratories worked collectively to evaluate 4 real-time RT-PCR (rRT-PCR) protocols targeting viral hemorrhagic septicemia virus (VHSV) being considered for deployment to a USA laboratory testing network. The protocols utilized previously published primers and probe sets developed for detection and surveillance of VHSV. All participating laboratories received and followed a standard operating protocol for extraction and for each of the rRT-PCR assays. Performance measures specifically evaluated included limit of detection (defined as the smallest amount of analyte in which 95% of the samples are classified as positive), analytical specificity, assay efficiency across genotype representatives, within- and between-plate variation within a laboratory, and variation between laboratories using the same platform, between platforms, and between software versions. This evaluation clearly demonstrated that the TaqMan®-based assay developed by Jonstrup et al. (2013; J Fish Dis 36:9-23) produced the most consistent analytical performance characteristics for detecting all genotypes of VHSV across the 8 participating laboratories.
【 授权许可】
Unknown
【 预 览 】
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RO201911300950143ZK.pdf | 732KB | download |