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Standard curves for Bundibugyo ebolavirus (BDBV) and mitochondrial circular DNA (mcirDNA) TaqMan RT-qPCR assays in singleplex and duplex formats. We generated standard curves for A) BDBV and B) the mcirDNA internal control assays using serial dilutions of synthetic RNA or DNA gene fragments. Data points show the mean and standard deviation of triplicate reactions. Solid lines indicate simple linear regressions, and dotted lines indicate the corresponding 95% confidence intervals. E denotes the RT-qPCR amplification efficiency.

Journal: medRxiv

Article Title: Rapid and portable reverse-transcription quantitative PCR assays for Bundibugyo ebolavirus detection

doi: 10.64898/2026.08.17.26360605

Figure Lengend Snippet: Standard curves for Bundibugyo ebolavirus (BDBV) and mitochondrial circular DNA (mcirDNA) TaqMan RT-qPCR assays in singleplex and duplex formats. We generated standard curves for A) BDBV and B) the mcirDNA internal control assays using serial dilutions of synthetic RNA or DNA gene fragments. Data points show the mean and standard deviation of triplicate reactions. Solid lines indicate simple linear regressions, and dotted lines indicate the corresponding 95% confidence intervals. E denotes the RT-qPCR amplification efficiency.

Article Snippet: Synthetic double stranded DNA gene fragments (Twist Bioscience, sequences provided in Table A4) targeting specific, unique regions of Z-EBOV (Viral Protein 24), S-EBOV (L segment, RNA-dependent RNA polymerase), BDBV (L segment, RNA-dependent RNA polymerase), and mcirDNA (Cytochrome C Oxidase Subunit III) were utilized as synthetic standards.

Techniques: Quantitative RT-PCR, Generated, Control, Standard Deviation, Amplification

Standard curves for Bundibugyo ebolavirus (BDBV), Zaire ebolavirus (Z-EBOV), Sudan ebolavirus (S-EBOV), and mitochondrial circular DNA (mcirDNA) TaqMan RT-qPCR assays in singleplex and four-target multiplex formats. We generated standard curves for A) BDBV, B) Z-EBOV, C) S-EBOV, and D) the mcirDNA internal control assays using serial dilutions of synthetic RNA or DNA gene fragments. Data points show the mean and standard deviation of triplicate reactions. Solid lines indicate simple linear regressions, and dotted lines indicate the corresponding 95% confidence intervals. E indicates the RT-qPCR amplification efficiency.

Journal: medRxiv

Article Title: Rapid and portable reverse-transcription quantitative PCR assays for Bundibugyo ebolavirus detection

doi: 10.64898/2026.08.17.26360605

Figure Lengend Snippet: Standard curves for Bundibugyo ebolavirus (BDBV), Zaire ebolavirus (Z-EBOV), Sudan ebolavirus (S-EBOV), and mitochondrial circular DNA (mcirDNA) TaqMan RT-qPCR assays in singleplex and four-target multiplex formats. We generated standard curves for A) BDBV, B) Z-EBOV, C) S-EBOV, and D) the mcirDNA internal control assays using serial dilutions of synthetic RNA or DNA gene fragments. Data points show the mean and standard deviation of triplicate reactions. Solid lines indicate simple linear regressions, and dotted lines indicate the corresponding 95% confidence intervals. E indicates the RT-qPCR amplification efficiency.

Article Snippet: Synthetic double stranded DNA gene fragments (Twist Bioscience, sequences provided in Table A4) targeting specific, unique regions of Z-EBOV (Viral Protein 24), S-EBOV (L segment, RNA-dependent RNA polymerase), BDBV (L segment, RNA-dependent RNA polymerase), and mcirDNA (Cytochrome C Oxidase Subunit III) were utilized as synthetic standards.

Techniques: Quantitative RT-PCR, Multiplex Assay, Generated, Control, Standard Deviation, Amplification

Journal: medRxiv

Article Title: Rapid and portable reverse-transcription quantitative PCR assays for Bundibugyo ebolavirus detection

doi: 10.64898/2026.08.17.26360605

Figure Lengend Snippet: Abridged Limit of detection (LOD) analysis for Bundibugyo ebolavirus (BDBV), Zaire ebolavirus (Z-EBOV), Sudan ebolavirus (S-EBOV), and mitochondrial circular DNA (mcirDNA) across TaqMan singleplex, duplex, four-target multiplex and SYBR Green RT-qPCR assay formats. We determined the 95% limit of detection (LOD 95 ) using synthetic RNA or DNA gene fragments quantified by digital PCR. We defined the LOD 95 as the lowest concentration detected in at least 95% of replicate reactions (n=21). Only results at the concentration meeting the LOD 95 criterion are shown; complete data are in Table A1.

Article Snippet: Synthetic double stranded DNA gene fragments (Twist Bioscience, sequences provided in Table A4) targeting specific, unique regions of Z-EBOV (Viral Protein 24), S-EBOV (L segment, RNA-dependent RNA polymerase), BDBV (L segment, RNA-dependent RNA polymerase), and mcirDNA (Cytochrome C Oxidase Subunit III) were utilized as synthetic standards.

Techniques: Multiplex Assay, SYBR Green Assay, Digital PCR, Concentration Assay

Standard curves for Bundibugyo ebolavirus (BDBV) and mitochondrial circular DNA (mcirDNA) TaqMan RT-qPCR assays in singleplex and duplex formats on the Mic qPCR cycler under rapid-cycling conditions. We generated standard curves for A) BDBV and B) mcirDNA internal control using serial dilutions of synthetic RNA or DNA gene fragments with the modified Mic qPCR protocol. Data points show the mean and standard deviation of triplicate reactions. Solid lines indicate simple linear regressions, and dotted lines indicate the corresponding 95% confidence interval. E denotes the RT-qPCR amplification efficiency.

Journal: medRxiv

Article Title: Rapid and portable reverse-transcription quantitative PCR assays for Bundibugyo ebolavirus detection

doi: 10.64898/2026.08.17.26360605

Figure Lengend Snippet: Standard curves for Bundibugyo ebolavirus (BDBV) and mitochondrial circular DNA (mcirDNA) TaqMan RT-qPCR assays in singleplex and duplex formats on the Mic qPCR cycler under rapid-cycling conditions. We generated standard curves for A) BDBV and B) mcirDNA internal control using serial dilutions of synthetic RNA or DNA gene fragments with the modified Mic qPCR protocol. Data points show the mean and standard deviation of triplicate reactions. Solid lines indicate simple linear regressions, and dotted lines indicate the corresponding 95% confidence interval. E denotes the RT-qPCR amplification efficiency.

Article Snippet: Synthetic double stranded DNA gene fragments (Twist Bioscience, sequences provided in Table A4) targeting specific, unique regions of Z-EBOV (Viral Protein 24), S-EBOV (L segment, RNA-dependent RNA polymerase), BDBV (L segment, RNA-dependent RNA polymerase), and mcirDNA (Cytochrome C Oxidase Subunit III) were utilized as synthetic standards.

Techniques: Quantitative RT-PCR, Generated, Control, Modification, Standard Deviation, Amplification

Standard curves for Bundibugyo ebolavirus (BDBV) and mitochondrial circular DNA (mcirDNA) TaqMan RT-qPCR assays in singleplex and duplex formats using lyophilized RT-qPCR mastermix on the QuantStudio 6 Flex and Mic qPCR cycler. We generated standard curves for A) BDBV on QuantStudio 6 Flex, B) the mcirDNA internal control on QuantStudio 6 Flex, C) BDBV on Mic qPCR cycler, and D) mcirDNA on the Mic qPCR cycler using serial dilutions of synthetic RNA gene fragments with lyophilized RT-qPCR mastermix. Data points show the mean and standard deviation of duplicate reactions. Solid lines indicate simple linear regressions, and dotted lines indicate the corresponding 95% confidence intervals. E denotes the RT-qPCR standard curve efficiency.

Journal: medRxiv

Article Title: Rapid and portable reverse-transcription quantitative PCR assays for Bundibugyo ebolavirus detection

doi: 10.64898/2026.08.17.26360605

Figure Lengend Snippet: Standard curves for Bundibugyo ebolavirus (BDBV) and mitochondrial circular DNA (mcirDNA) TaqMan RT-qPCR assays in singleplex and duplex formats using lyophilized RT-qPCR mastermix on the QuantStudio 6 Flex and Mic qPCR cycler. We generated standard curves for A) BDBV on QuantStudio 6 Flex, B) the mcirDNA internal control on QuantStudio 6 Flex, C) BDBV on Mic qPCR cycler, and D) mcirDNA on the Mic qPCR cycler using serial dilutions of synthetic RNA gene fragments with lyophilized RT-qPCR mastermix. Data points show the mean and standard deviation of duplicate reactions. Solid lines indicate simple linear regressions, and dotted lines indicate the corresponding 95% confidence intervals. E denotes the RT-qPCR standard curve efficiency.

Article Snippet: Synthetic double stranded DNA gene fragments (Twist Bioscience, sequences provided in Table A4) targeting specific, unique regions of Z-EBOV (Viral Protein 24), S-EBOV (L segment, RNA-dependent RNA polymerase), BDBV (L segment, RNA-dependent RNA polymerase), and mcirDNA (Cytochrome C Oxidase Subunit III) were utilized as synthetic standards.

Techniques: Quantitative RT-PCR, Generated, Control, Standard Deviation