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ResultsColumnDefinitionPossible Entries
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Sample #Known ValueSample TypeSample SourceKnown ValueKnown UnitsResult ValueResult UnitsResult InterpretationNotesSample #Unique number for sampleNumeric (1-number of samples tested)
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NumericTextTextTextNumericTextNumericTextTextTextKnown ValueSAR-CoV-2 sample statusPositive, Negative
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Known ValueIf Clinical sample what is the result from the clinical test; If contrived what amount of virus is used in units of copies per uLNumeric
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Blinded real saliva samples, spiked with zeptometrix inactivated virions into raw sample before any processing
Known UnitsIf Clinical sample what is the units from the clinical test; If contrived this must be copies/uLText (Ct, copies (of virus in reaction))
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Run#39-1NegativeReal UnspikedSalivaNegativeVisual binaryCorrectAs1e and N2 Primer Set ComboResult ValueSample up to 1 decimal pointNumeric
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Run#39-2NegativeReal UnspikedSalivaNegativeVisual binaryCorrectAs1e and N2 Primer Set ComboResult UnitsUnits used for reportingText (Ct, fluorescence)
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Run#39-3Positive
Real, Raw Zepto Spiked
Saliva5E+03virions/mlPositiveVisual binaryCorrectAs1e and N2 Primer Set ComboResult InterpretationBased off of this result would you declare this positive or negativePositive, Negative
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Run#39-4NegativeReal UnspikedSalivaNegativeVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#39-5Positive
Real, Raw Zepto Spiked
Saliva2E+04virions/mlPositiveVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#39-6NegativeReal UnspikedSalivaNegativeVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#39-7NegativeReal UnspikedSalivaNegativeVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#39-8Positive
Real, Raw Zepto Spiked
Saliva1E+04virions/mlPositiveVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#39-9NegativeReal UnspikedSalivaNegativeVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#39-10NegativeReal UnspikedSalivaNegativeVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#39-NTCNegativeControlPBSNegativeVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#39-TWCPositiveContrivedSaliva2,500copies (est)PositiveVisual binaryCorrectAs1e and N2 Primer Set Combo
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Twist LOD run on 2ml volume of inactivated sample, Twist positive control RNA spiked after sample inactivation before purification, same results in duplicate
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Run#37-8KPositiveContrivedSaliva4,000copies (est)PositiveVisual binaryAt or above LODAs1e and N2 Primer Set Combo
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Run#37-4KPositiveContrivedSaliva2,000copies (est)PositiveVisual binaryAt or above LODAs1e and N2 Primer Set Combo
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Run#37-2KPositiveContrivedSaliva1,000copies (est)PositiveVisual binaryAt or above LODAs1e and N2 Primer Set Combo
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Run#37-1KPositiveContrivedSaliva500copies (est)NegativeVisual binaryBelow LODAs1e and N2 Primer Set Combo
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Run#37-0NegativeControlSaliva0NegativeVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#37-0Positive
Control (Internal RNAseP)
SalivaHIGHPositiveVisual binaryCorrectRNAseP Primers
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Run#37-NTCNegativeControlPBS0NegativeVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#37-NTCNegative
Control (Internal RNAseP)
PBS0NegativeVisual binaryCorrectRNAseP Primers
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Real Sample run on individual for a screen, 2 nasal samples split and 25K zeptos spiked into 2.5ml
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Run#86-1A?RealNasalNegativeVisual binary
No Finding of Potential Clinical Significance
As1e and N2 Primer Set Combo
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Run#86-1BPositive
Real, Raw Zepto Spiked
Nasal1E+04virions/mlPositiveVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#86-2ANegativeRealNasalNegativeVisual binary
No Finding of Potential Clinical Significance
As1e and N2 Primer Set Combo
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Run#86-2BPositive
Real, Raw Zepto Spiked
Nasal1E+04virions/mlPositiveVisual binaryCorrectAs1e and N2 Primer Set Combo
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Run#86-3NegativeRealNasalNegativeVisual binary
No Finding of Potential Clinical Significance
As1e and N2 Primer Set Combo
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Run#86-4NegativeRealNasalNegativeVisual binary
No Finding of Potential Clinical Significance
As1e and N2 Primer Set Combo
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Run#86-5?RealSalivaNegativeVisual binary
No Finding of Potential Clinical Significance
As1e and N2 Primer Set Combo
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Run#86-NTCNegativeControlPBSNegativeVisual binaryCorrectAs1e and N2 Primer Set Combo
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These are a few example runs, we have done over 100 runs in the last 2 months, 1000+ LAMP reactions
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Many have been chacterization and troubleshooting runs, as we've moved labs 3 times
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We now have great exclusive use lab space at MBC Biolabs with:
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a dedicated room with 2 biosafety cabinets where we will run the silica purification on inactivated samples
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4 lab benches where we'll install 2 PCR hood for setting up LAMP reactions
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1 5' chemical fume hood for preparing stock solutions invloving acids and bases (Inactivation Solution, Binding Solution and Glass Milk)
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We have access to a GloMAX plate reader and ABI Quanstudio7 qPCR machine which we intend to use for comparison and characterization, and perhaps as part of our screening service offering
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We have done many successful runs spiking Zeptometrix virions into raw samples at 1e4/ml level.
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The challenge is not getting good results on one run, but setting up a system to run continously under control, especially eliminating failures due to contamination.
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We've moved to single use RNA controls, and aliquoting of many components in specific volumes in preparation for pilot runs.
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