Detection Limit Determination using the Four-Parameter Logistic Model for the Ultrasensitive Detection of Vibrio cholerae DNA with Polystyrene-coacrylic Acid Composite Nanospheres
Keywords:Detection limit, Four-Parameter Logistic Model, Pooled Standard Deviation, Biosensor, Vibrio cholerae
Standard laboratory techniques for isolating and identifying V. cholerae can take up to three days and need a well-equipped laboratory with highly experienced personnel. If cholera epidemics go unnoticed for a lengthy period of time, inadequate public health measures, disease spread, and increased death and morbidity are all possible. Cholera cases must be detected and confirmed as soon as feasible using sustainable and precise approaches in order to provide necessary support. Biochemical diagnostic techniques for this pathogen, such as antigen binding, rely on biomolecular interactions as its diagnostic modality, resulting in more complex calibration curves. Additionally, these tests frequently employ sigmoidal curves. When there is asymmetry, a logistic (5PL) or logistic (4PL) curve may be the best representation of a separate sigmoidal relationship. It is possible that linearizing an otherwise nonlinear connection by log transformation disrupts the curve's error structure, having the opposite effect of reducing or even eliminating error in the relationship. The aim of this study is the remodel the data from a calibration curve for the detection of Vibrio cholerae DNA by means of polystyrene-coacrylic acid composite nanospheres using the standard 4-PL model and to determine the Limits of Detection (LOD) based on two methods; the classical definition of limits of detection method and the method based on pooled standard deviation (PSD). The LOD value obtained through the 4PL modelling exercise based on the classical method was 0.255 pM (95% confidence interval of 0.167 to 0.379) while the PSD method yielded an LOD value of 0.035 pM (95% confidence interval of 0.011 to 0.067), which indicates that the PSD method was superior. The classical method was higher than the rough estimation employed originally with an LOD value of 0.125 pM. The use of the 4PL method based on the PSD method was more reliable and robust in estimating the LOD values.
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