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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
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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