Ni assays troubleshooting analysis of curve fitting
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NI Assays: Troubleshooting & Analysis of Curve Fitting. Graph 1: Standard clinical isolate Good NA activity S shaped curve (observed points) IC50 in expected range for susceptible B Good fit of calculated curve to observed points Conclusion: VALID sensitive isolate.

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NI Assays: Troubleshooting & Analysis of Curve Fitting

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Ni assays troubleshooting analysis of curve fitting

NI Assays: Troubleshooting & Analysis of Curve Fitting

Graph 1:Standard clinical isolate

  • Good NA activity

  • S shaped curve (observed points)

  • IC50 in expected range for susceptible B

  • Good fit of calculated curve to observed points

    Conclusion: VALID sensitive isolate


Ni assays troubleshooting analysis of curve fitting1

NI Assays: Troubleshooting & Analysis of Curve Fitting

Graph 2:Known resistant mutant

  • Low NA activity

  • Known resistant virus

  • High IC50

  • S shaped curve, but displaced right because of drug inhibition

  • Good fit curve to observed points

    Conclusion: VALID resistant isolate


Ni assays troubleshooting analysis of curve fitting2

NI Assays: Troubleshooting & Analysis of Curve Fitting

Graph 3:Clinical isolate with low IC50

  • Low NA activity

  • Poor fit observed points to calculated curve

  • IC50 value low

    Conclusion: Probably isolate with normal IC50 → REPEAT


Ni assays troubleshooting analysis of curve fitting3

NI Assays: Troubleshooting & Analysis of Curve Fitting

Graph 4:Clinical isolate with high IC50

  • Low NA activity

  • Poor fit observed points to curve

  • IC50 displaced to right

  • High IC50

    Conclusion: Probably resistant isolate REPEAT to check


Ni assays troubleshooting analysis of curve fitting4

NI Assays: Troubleshooting & Analysis of Curve Fitting

Graph 5:Clinical isolate with low IC50

  • Good NA activity

  • Poor fit observed values to curve especially at high drug concentrations

  • Non sigmoidal shape of observed curve

  • Low IC50

    Conclusion: Probable error drug dilution for normal iC50 invalid → REPEAT


Ni assays troubleshooting analysis of curve fitting5

NI Assays: Troubleshooting & Analysis of Curve Fitting

Graph 6:Clinical isolate with high IC50

  • Low NA activity

  • Non sigmoidal observed points

  • High IC50

  • Bimodal distribution observed points

  • Poor fit between observed points and curve

    Conclusion: Possible resistant but technical errors → REPEAT


Ni assays troubleshooting analysis of curve fitting6

NI Assays: Troubleshooting & Analysis of Curve Fitting

Graph 7:Clinical isolate with normal IC50

  • Good NA activity

  • Increase in drug concentration associated with increase NA activity (Biologically implausible)

  • Non sigmoidal observed points

  • Poor fit observed points and curve

    Conclusion: Invalid analysis but probably normal IC50. Technical errors → REPEAT


Ni assays troubleshooting analysis of curve fitting7

NI Assays: Troubleshooting & Analysis of Curve Fitting

Graph 8:Clinical isolate with low IC50

  • Low NA activity

  • Increase NA activity with drug concentration (Biologically implausible)

  • Bimodal distribution observed points

  • Poor fit observed points to curve

  • Low IC50

    Conclusion: INVALID analysis → REPEAT as probably normal IC50


Ni assays troubleshooting analysis of curve fitting8

NI Assays: Troubleshooting & Analysis of Curve Fitting

Graph 9:Clinical isolate with low IC50

  • Good NA activity

  • NA activity increases with drug concentration

  • Bimodal distribution observed points

  • Poor fit curves and observed points

  • Low IC50

    Conclusion: Invalid analysis → REPEAT as probably normal IC50


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