Point-of-Care Disease Screening in Primary Care Using Saliva: A Biospectroscopy Approach for Lung Cancer and Prostate Cancer

Abstract

Saliva is a largely unexplored liquid biopsy that can be readily obtained noninvasively. Not dissimilar to blood plasma or serum, it contains a vast array of bioconstituents that may be associated with the absence or presence of a disease condition. Given its ease of access, the use of saliva is potentially ideal in a point-of-care screening or diagnostic test. Herein, we developed a swab "dip" test in saliva obtained from consenting patients participating in a lung cancer-screening programme being undertaken in north-west England. A total of 998 saliva samples (31 designated as lung-cancer positive and 17 as prostate-cancer positive) were taken in the order in which they entered the clinic (i.e., there was no selection of participants) during the course of this prospective screening programme. Samples (sterile Copan blue rayon swabs dipped in saliva) were analysed using attenuated total reflection Fourier-transform infrared (ATR-FTIR) spectroscopy. In addition to unsupervised classification on resultant infrared (IR) spectra using principal component analysis (PCA), a range of feature selection/extraction algorithms were tested. Following preprocessing, the data were split between training (70% of samples, 22 lung-cancer positive versus 664 other) and test (30% of samples, 9 lung-cancer positive versus 284 other) sets. The training set was used for model construction and the test set was used for validation. The best model was the PCA-quadratic discriminant analysis (QDA) algorithm. This PCA-QDA model was built using 8 PCs (90.4% of explained variance) and resulted in 93% accuracy for training and 91% for testing, with clinical sensitivity at 100% and specificity at 91%. Additionally, for prostate cancer patients amongst the male cohort (n = 585), following preprocessing, the data were split between training (70% of samples, 12 prostate-cancer positive versus 399 other) and test (30% of samples, 5 prostate-cancer positive versus 171 other) sets. A PCA-QDA model, again the best model, was built using 5 PCs (84.2% of explained variance) and resulted in 97% accuracy for training and 93% for testing, with clinical sensitivity at 100% and specificity at 92%. These results point to a powerful new approach towards the capability to screen large cohorts of individuals in primary care settings for underlying malignant disease.

Keywords: ATR-FTIR spectroscopy; cancer screening; chemometrics; saliva; swab; “dip” test.

Figure 1

Mid-infrared (IR) spectra derived using ATR-FTIR spectroscopy. (A) Raw IR spectra for all sample in each group (lung cancer or other conditions [OTHER]); (B) average raw IR spectra for each group (lung cancer or other conditions [OTHER]). (C) Preprocessed (SG 2nd derivative followed by vector normalisation) IR spectra for all samples in each group (lung cancer or other conditions [OTHER]); (D) average preprocessed IR spectra for each group (lung cancer or other conditions [OTHER]).

Figure 2

Difference-between-mean (DBM) spectrum and PCA loadings on the 8 PCs used to build the PCA-QDA model. The “TOTAL” stands for the sum of coefficients in the 8 PCs.

Figure 3

Mid-infrared (IR) spectra derived using ATR-FTIR spectroscopy. (A) Raw IR spectra for all samples in each group (prostate cancer [P-CA] or other conditions for male patients [OTHER]); (B) average raw IR spectra for each group (prostate cancer [P-CA] or other conditions for male patients [OTHER]). (C) Preprocessed (SG 2nd derivative followed by vector normalisation) IR spectra for all samples in each group (prostate cancer [P-CA] or other conditions for male patients [OTHER]); (D) average preprocessed IR spectra for each group (prostate cancer [P-CA] or other conditions for male patients [OTHER]).

Figure 4

Difference-between-mean (DBM) spectrum and PCA loadings on the 5 PCs used to build the PCA-QDA model. The “TOTAL” stands for the sum of coefficients in the 5 PCs.