Detection and surveillance of viral hemorrhagic septicemia virus using real-time RT-PCR. I. Initial comparison of four protocols
| Autor: | Robert Kim, Janet V. Warg, Andrew E. Goodwin, Mohamed Faisal, Geoffrey H. Groocock, Monica M Reising, Rodman G. Getchell, Nicholas B. D. Phelps, Isaac Standish, Cem Giray, Angela Cruz, Yan Zhang, Kathy Toohey-Kurth, Travis Clement, Emily R. Cornwell, Gwenn E. Merry |
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| Rok vydání: | 2014 |
| Předmět: |
Protocol (science)
Analyte biology Genotype Reverse Transcriptase Polymerase Chain Reaction Fishes Reproducibility of Results Hemorrhagic septicemia Aquatic Science biology.organism_classification Real-Time Polymerase Chain Reaction Laboratory testing Virology Sensitivity and Specificity Virus Novirhabdovirus Real-time polymerase chain reaction Population Surveillance Hemorrhagic Septicemia Viral Animals Viral hemorrhagic septicemia Ecology Evolution Behavior and Systematics |
| Zdroj: | Diseases of aquatic organisms. 111(1) |
| ISSN: | 0177-5103 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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