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Pre-processing is a crucial step in analyzing spectra from Fourier transform infrared (FTIR) spectroscopy because it can reduce unwanted noise and enhance system performance. Here, we present the results of pre-processing technique optimization to facilitate the detection of pepper yellow leaf curl virus (PYLCV)-infected chilli plants using FTIR spectroscopy. Optimization of a range of pre-processing techniques was undertaken, namely baseline correction, normalization (standard normal variate, vector, and min–max), and de-noising (Savitzky-Golay (SG) smoothing, 1st and 2 derivatives). The pre-processing was applied to the mid-infrared spectral range (4000 – 400 cm−1) and the biofingerprint region (1800 – 900 cm−1) then the discrete wavelet transform (DWT) was used for dimension reduction. The pre-processed data were then used as an input for classification using a multilayer perceptron neural network, a support vector machine, and linear discriminant analysis. The pre-processing method with the highest classification model accuracy was selected for the further use in the processing. It was seen that only the SG 1st derivative method applied to both wavenumber ranges could produce 100% accuracy. This result was supported by principal component analysis clustering. Thus, we have demonstrated that by using the right pre-processing technique, classification success can be increased, and the process simplified by optimization and minimization of the technique used.