Exploring pre-analytical factors for the optimisation of serum diagnostics: Progressing the clinical utility of ATR-FTIR spectroscopy

Cameron, James M.; Butler, Holly J.; Anderson, David J.; Christie, Loren; Confield, Lily Rose; Spalding, Katie E.; Finlayson, Duncan; Murray, Stuart; Panni, Zanib; Rinaldi, Christopher A.; Sala, Alexandra; Theakstone, Ashton G.; Baker, Matthew J.

doi:10.1016/j.vibspec.2020.103092

Cited by 16 publications

(18 citation statements)

References 48 publications

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“…Likewise, FTIR can detect and stratify brain malignancies through the analysis of resected tissue sections, mainly focused on transmission techniques [33][34][35][36]. On the other hand, attenuated total reflection (ATR)-FTIR is well-suited to biological fluids, such as blood serum [37,38]. The technique provides for the qualitative interrogation of all infrared active macromolecular constituents of blood serum, and it is well established that biomolecular imbalances in biofluids can give an indication of disease states [28].…”

Section: Glioma Entity Who Grade Idh1 Mutationmentioning

confidence: 99%

Interrogation of IDH1 Status in Gliomas by Fourier Transform Infrared Spectroscopy

Cameron

Conn

Rinaldi

et al. 2020

Cancers

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Mutations in the isocitrate dehydrogenase 1 (IDH1) gene are found in a high proportion of diffuse gliomas. The presence of the IDH1 mutation is a valuable diagnostic, prognostic and predictive biomarker for the management of patients with glial tumours. Techniques involving vibrational spectroscopy, e.g., Fourier transform infrared (FTIR) spectroscopy, have previously demonstrated analytical capabilities for cancer detection, and have the potential to contribute to diagnostics. The implementation of FTIR microspectroscopy during surgical biopsy could present a fast, label-free method for molecular genetic classification. For example, the rapid determination of IDH1 status in a patient with a glioma diagnosis could inform intra-operative decision-making between alternative surgical strategies. In this study, we utilized synchrotron-based FTIR microanalysis to probe tissue microarray sections from 79 glioma patients, and distinguished the positive class (IDH1-mutated) from the IDH1-wildtype glioma, with a sensitivity and specificity of 82.4% and 83.4%, respectively. We also examined the ability of attenuated total reflection (ATR)-FTIR spectroscopy in detecting the biomolecular events and global epigenetic and metabolic changes associated with mutations in the IDH1 enzyme, in blood serum samples collected from an additional 72 brain tumour patients. Centrifugal filtration enhanced the diagnostic ability of the classification models, with balanced accuracies up to ~69%. Identification of the molecular status from blood serum prior to biopsy could further direct some patients to alternative treatment strategies.

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Section: Glioma Entity Who Grade Idh1 Mutationmentioning

confidence: 99%

Interrogation of IDH1 Status in Gliomas by Fourier Transform Infrared Spectroscopy

Cameron

Conn

Rinaldi

et al. 2020

Cancers

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“…Biofluids should be stored in a −80°C freezer in cryovials or Eppendorf tubes after collection from pathology laboratories or biobanks. The period of time concerning long‐term storage of biofluid samples does not appear to influence acquired spectral data as established by previous work exploring pre‐analytical factors with human serum samples [123]. Whole biofluid samples are most often used, but some studies may involve dilution [86], filtration or centrifugation (eg, to examine the low molecular weight fraction of serum) [68, 87].…”

Section: Methodsmentioning

confidence: 99%

“…Similarly, whole blood samples are commonly subjected to centrifugation processes to extract serum or plasma constituents prior to clinical testing. Differences in centrifugation speeds and time intervals between protocols have not been found to influence spectral data, although it is important to highlight that minor spectral differences have been observed between different serum collection tube types [123]. Volumes of required sample are dependent on mode of spectral collection.…”

Section: Methodsmentioning

confidence: 99%

Fourier‐transform infrared spectroscopy of biofluids: A practical approach

Theakstone

Rinaldi

Butler³

et al. 2021

Transl Biophotonics

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Biofluid spectroscopy is an emerging technology in the field of clinical investigation, providing a simple way to extract diagnostic and observational information from easy to acquire samples. Infrared spectroscopy is well suited to analyse a large range of biofluid samples, including blood and its derivatives, due to flexible sampling modes and high sensitivity to subtle biological changes. As the technology advances towards the clinic, factors influencing successful clinical translation are becoming apparent. Here, we provide a tutorial for effective biofluid spectroscopy study design, discussing sample and instrument parameters, as well as clinical considerations. The aim is to present the current understanding of clinical translation in the field of biofluid spectroscopy, and to facilitate other clinical applications to advance to the clinic.

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“…Aliquots of 1 μL of serum (1:3 in ultrapure water) were deposited in platinum sample holder and dried at 80% relative moisture under a desicator with NaCl saturated solution in order to avoid cofee ring effect and obtain a homogeneous biofilm [26]. The final droplet presented an average radius of 1000 +/-150 mm.…”

Section: Fourier-transform Infrared Absorption (Ftir) Spectroscopymentioning

confidence: 99%

“…Then only a single patient could be tested at once. This process demands usually > 5 hours[26]. It is not well reported whether the proteins degradation over this time would impact on quality of diagnosis.…”

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confidence: 99%

Micro-Fourier-Transform Infrared Reflectance Spectroscopy as Tool for Probing IgG Glycosylation in COVID-19 Patients

Bandeira

Madureira

Rossi

et al. 2021

Preprint

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It has been reported that patients diagnosed with COVID-19 become critically ill primarily around the time of activation of the adaptive immune response. However the role of antibodies in the worsening of disease is not obvious. Higher titers of anti-spike immunoglobulin IgG1 associated to low fucosylation of the antibody Fc tail have been associated to excessive inflammatory response. In contrast it has been also reported in literature that NP-, S-, RBD- specific IgA, IgG, IgM are not associated with SARS-CoV-2 viral load, indicating that there is no obvious correlation between antibody response and viral antigen detection. In the present work the micro-Fourier-Transform Infrared reflectance spectroscopy (micro-FTIR) was employed to investigate blood serum samples of healthy and COVID-19 (mild or oligosynthomatic) individuals (82 healthcare workers volunteers in “Instituto de Infectologia Emilio Ribas”, São Paulo, Brazil). The molecular-level-sensitive, multiplexing quantitative and qualitative FTIR data probed on 1 mL of dryed biofluidwas compared to Signal-to-Cutoff index of chemiluminescent immunoassays CLIA and ELISA (IgG antibodies against SARS-CoV-2). Our main result indicated that 1702-1785 cm-1 spectral window (carbonyl C=O vibration) appeared to be a spectral marker of the degree of IgG glycosylation, allowing to probe distinctive subpopulations of COVID-19 patients, depending on their degree of severity. The specificity was 87.5 % while the detection rate of true positive was 100%. The computed area under the receiver operating curve was equivalent to CLIA, ELISA and other ATR-FTIR methods (> 0.85). In summary, overall discrimination of healthy and COVID-19 individuals and severity prediction as well could also be potentially implemented using micro-FTIR reflectance spectroscopy on blood serum samples. Considering the minimal and reagent-free sample preparation procedures combined to fast (few minutes) outcome of FTIR we can state that this technology is very suitable for fast screening of immune response of individuals with COVID-19. It would be an important tool in prospective studies, helping investigate the physiology of the asymptomatic, oligosymptomatic, or severe individuals and measure the extension of infection dissemination in patients.

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Exploring pre-analytical factors for the optimisation of serum diagnostics: Progressing the clinical utility of ATR-FTIR spectroscopy

Cited by 16 publications

References 48 publications

Interrogation of IDH1 Status in Gliomas by Fourier Transform Infrared Spectroscopy

Interrogation of IDH1 Status in Gliomas by Fourier Transform Infrared Spectroscopy

Fourier‐transform infrared spectroscopy of biofluids: A practical approach

Micro-Fourier-Transform Infrared Reflectance Spectroscopy as Tool for Probing IgG Glycosylation in COVID-19 Patients

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