Duplex droplet digital PCR enables simultaneous quantification of algal giant virus DSLLAV1 and virophage DSLV8 in natural and laboratory samples.
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| Format: | Artículo científico |
| Langue: | en |
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Journal of virological methods
2026
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| _version_ | 1868266144187547649 |
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| author | Chu, Ting Yu, Jiabei Wang, Qinran Hu, Chen Chen, Lanming Yu, Yongxin Wang, Yongjie |
| author_facet | Chu, Ting Yu, Jiabei Wang, Qinran Hu, Chen Chen, Lanming Yu, Yongxin Wang, Yongjie Chu, Ting Yu, Jiabei Wang, Qinran Hu, Chen Chen, Lanming Yu, Yongxin Wang, Yongjie |
| collection | PubMed - marine biology |
| contents | Duplex droplet digital PCR enables simultaneous quantification of algal giant virus DSLLAV1 and virophage DSLV8 in natural and laboratory samples. Chu, Ting Yu, Jiabei Wang, Qinran Hu, Chen Chen, Lanming Yu, Yongxin Wang, Yongjie Sensitivity and Specificity Virophages Lakes DNA Primers Polymerase Chain Reaction Giant Viruses Real-Time Polymerase Chain Reaction Genome, Viral Cell-virus-virophage (CVv) systems involve virophages parasitizing giant viruses within eukaryotic hosts, forming unique virus-virus interactions with complex ecological implications. However, quantitative tools for studying such systems-particularly in freshwater algae-remain limited. In this study, we developed and optimized a duplex droplet digital PCR (ddPCR) assay to simultaneously detect and quantify Dishui Lake Large Algal Virus 1 (DSLLAV1), a Mimiviridae-like algal giant virus, and its associated Dishui Lake virophage 8 (DSLV8) in the Dishui Lake ecosystem. Target-specific primers and TaqMan probes were designed based on viral genomic sequences, and assay conditions were optimized for annealing temperature, primer/probe concentrations, and droplet separation. The established assay demonstrated high specificity and sensitivity, with detection limits of 0.13 and 0.16 copies/µL for DSLLAV1 and DSLV8, respectively. The method outperformed qPCR in sensitivity and maintained stability across environmental and infection-derived samples. This ddPCR method provides a robust platform for monitoring virus-virophage dynamics and offers new opportunities for investigating the ecological and evolutionary roles of CVv systems in aquatic environments. |
| format | Artículo científico |
| id | pubmed_41047082 |
| institution | PubMed |
| language | en |
| publishDate | 2026 |
| publisher | Journal of virological methods |
| record_format | pubmed |
| spellingShingle | Duplex droplet digital PCR enables simultaneous quantification of algal giant virus DSLLAV1 and virophage DSLV8 in natural and laboratory samples. Chu, Ting Yu, Jiabei Wang, Qinran Hu, Chen Chen, Lanming Yu, Yongxin Wang, Yongjie Sensitivity and Specificity Virophages Lakes DNA Primers Polymerase Chain Reaction Giant Viruses Real-Time Polymerase Chain Reaction Genome, Viral Duplex droplet digital PCR enables simultaneous quantification of algal giant virus DSLLAV1 and virophage DSLV8 in natural and laboratory samples. Chu, Ting Yu, Jiabei Wang, Qinran Hu, Chen Chen, Lanming Yu, Yongxin Wang, Yongjie Sensitivity and Specificity Virophages Lakes DNA Primers Polymerase Chain Reaction Giant Viruses Real-Time Polymerase Chain Reaction Genome, Viral Cell-virus-virophage (CVv) systems involve virophages parasitizing giant viruses within eukaryotic hosts, forming unique virus-virus interactions with complex ecological implications. However, quantitative tools for studying such systems-particularly in freshwater algae-remain limited. In this study, we developed and optimized a duplex droplet digital PCR (ddPCR) assay to simultaneously detect and quantify Dishui Lake Large Algal Virus 1 (DSLLAV1), a Mimiviridae-like algal giant virus, and its associated Dishui Lake virophage 8 (DSLV8) in the Dishui Lake ecosystem. Target-specific primers and TaqMan probes were designed based on viral genomic sequences, and assay conditions were optimized for annealing temperature, primer/probe concentrations, and droplet separation. The established assay demonstrated high specificity and sensitivity, with detection limits of 0.13 and 0.16 copies/µL for DSLLAV1 and DSLV8, respectively. The method outperformed qPCR in sensitivity and maintained stability across environmental and infection-derived samples. This ddPCR method provides a robust platform for monitoring virus-virophage dynamics and offers new opportunities for investigating the ecological and evolutionary roles of CVv systems in aquatic environments. |
| title | Duplex droplet digital PCR enables simultaneous quantification of algal giant virus DSLLAV1 and virophage DSLV8 in natural and laboratory samples. |
| topic | Sensitivity and Specificity Virophages Lakes DNA Primers Polymerase Chain Reaction Giant Viruses Real-Time Polymerase Chain Reaction Genome, Viral |
| url | https://pubmed.ncbi.nlm.nih.gov/41047082/ |