Non-invasive Disease Diagnosis Research Articles

GFP-SCFV: expression and possible applications as a tool for experimental investigations of atherosclerosis.

Experimental studies on atherosclerosis are crucial for investigating its pathophysiology, defining new therapeutic targets, and developing new drugs and diagnostic tools. Thus, many imaging markers have been developed and introduced in experimental studies. The main advantage of these new tools is that they allow the noninvasive diagnosis of atherosclerotic vascular disease. Here, we describe the cloning, expression, purification, and stabilization of a chimeric protein specifically designed to probe cells and tissues for the presence of LDL(-), a relevant marker of atherosclerosis. The DNA sequence that encodes the anti-LDL(-) scFv, previously obtained from a hybridoma secreting an anti-LDL(-) monoclonal antibody, was inserted into the bacterial vector pET-28a(+) in tandem with a DNA sequence encoding GFP. The recombinant protein was expressed in high yields in E. coli as inclusion bodies. The applicability of GFP-scFv was assessed by ELISA, which determined its affinity for LDL(-) and confocal microscopy, that showed macrophage uptake of the protein along with LDL(-). In conclusion, our data suggest that the anti-LDL(-) GFP-scFv chimeric protein could be useful in studies on atherogenesis as well as for developing diagnostic tools for atherosclerosis.

Characterization of Exhaled Particles from the Human Lungs in Airway Obstruction

Human breath contains small particles that might be useful for the noninvasive diagnosis of lung disease. In this study, the impact of airway obstruction on particle emission was investigated. Particle number flux and particle size distribution were measured for healthy nonsmokers (n=16), healthy smokers (n=13), patients with chronic obstructive pulmonary disease (n=28, GOLD stage I-IV), and patients with asthma before and after methacholine challenge (n=10). The measurements were carried out using a condensation nucleus counter (TSI 3760) and a laser spectrometer (PMT LASAIR II-110). Particle number per breath showed high intrasubject reproducibility. However, there was a large intersubject variability in the number of emitted particles on the order of two magnitudes, with no influence of airway obstruction on emission level. Methacholine-induced airway obstruction, in subjects with allergic asthma, did not change the number of exhaled particles, when compared with prechallenge values. For the droplet size distribution averaged per breath, there was no difference between healthy subjects and subjects with airway obstruction. Airway obstruction does not change the number flux or size distribution of particles in exhaled breath. The high intersubject variability of particle emission supports the concept of online determination of aerosol properties (primarily number flux, during exhaled breath) during breath condensate sampling to properly normalize the results of biochemical analysis. As high dilution and variable dilution are the main challenges of biomarker assessment in exhaled breath condensate, this normalization procedure would significantly add to the value of the technique.

Fingerprinting breast cancer vs. normal mammary cells by mass spectrometric analysis of volatiles.

There is increasing interest in the development of noninvasive diagnostic methods for early cancer detection, to improve the survival rate and quality of life of cancer patients. Identification of volatile metabolic compounds may provide an approach for noninvasive early diagnosis of malignant diseases. Here we analyzed the volatile metabolic signature of human breast cancer cell lines versus normal human mammary cells. Volatile compounds in the headspace of conditioned culture medium were directly fingerprinted by secondary electrospray ionization-mass spectrometry. The mass spectra were subsequently treated statistically to identify discriminating features between normal vs. cancerous cell types. We were able to classify different samples by using feature selection followed by principal component analysis (PCA). Additionally, high-resolution mass spectrometry allowed us to propose their chemical structures for some of the most discriminating molecules. We conclude that cancerous cells can release a characteristic odor whose constituents may be used as disease markers.

Application of gas chromatography mass spectrometry (GC–MS) in conjunction with multivariate classification for the diagnosis of gastrointestinal diseases

Gastrointestinal diseases such as irritable bowel syndrome, Crohn’s disease (CD) and ulcerative colitis are a growing concern in the developed world. Current techniques for diagnosis are often costly, time consuming, inefficient, of great discomfort to the patient, and offer poor sensitivities and specificities. This paper describes the development and evaluation of a new methodology for the non-invasive diagnosis of such diseases using a combination of gas chromatography mass spectrometry (GC–MS) and chemometrics. Several potential sample matrices were tested: blood, breath, faeces and urine. Faecal samples provided the only statistically significant results, providing discrimination between CD and healthy controls with an overall classification accuracy of 85 % (78 % specificity; 93 % sensitivity). Differentiating CD from other diseases proved more challenging, with overall classification accuracy dropping to 79 % (83 % specificity; 68 % sensitivity). This diagnostic performance compares well with the gold standard technique of colonoscopy, suggesting that GC–MS may have potential as a non-invasive screening tool.

DIMETER: A Haptic Master Device for Tremor Diagnosis in Neurodegenerative Diseases

In this study, a device based on patient motion capture is developed for the reliable and non-invasive diagnosis of neurodegenerative diseases. The primary objective of this study is the classification of differential diagnosis between Parkinson's disease (PD) and essential tremor (ET). The DIMETER system has been used in the diagnoses of a significant number of patients at two medical centers in Spain. Research studies on classification have primarily focused on the use of well-known and reliable diagnosis criteria developed by qualified personnel. Here, we first present a literature review of the methods used to detect and evaluate tremor; then, we describe the DIMETER device in terms of the software and hardware used and the battery of tests developed to obtain the best diagnoses. All of the tests are classified and described in terms of the characteristics of the data obtained. A list of parameters obtained from the tests is provided, and the results obtained using multilayer perceptron (MLP) neural networks are presented and analyzed.

Capillary electrophoresis with laser-induced fluorescence detection of main polyamines and precursor amino acids in saliva

Abstract A hollow-fiber liquid-phase microextraction (HF-LPME) method has been developed for the purification and preconcentration of biogenic polyamines and their precursor amino acids in human saliva. Putrescine (Put), cadaverine (Cad), spermidine (Spe), ornithine (Orn), lysine (Lys), and arginine (Arg) were determined by the CE-LIF detection after microextraction. Several factors that affect extraction efficiency, separation, and detection were investigated. Under the optimum conditions, six analytes could achieve baseline separation within 30 min, exhibiting a linear calibration at three orders of magnitude ( r 2 > 0.998); the obtained enrichment factors of HF-LPME were between 19 (for Orn) and 218 (for Cad), and the LODs were in the range of 0.0072–0.26 nmol/L. The proposed HF-LPME/CE-LIF method has been successfully applied for the sensitive analyses of the real-world saliva samples collected from healthy volunteers and different patients with oral diseases, providing a potential method for primary non-invasive diagnosis of some oral diseases.

Macular diseases: epiretinal membrane.

Epiretinal membrane (ERM) is a relatively common macular disease, which forms along the surface of the internal limiting membrane of the retina. Currently, microincision vitrectomy surgery (MIVS) has been confirmed as a minimally invasive and very safe modality of treatment that is useful for improving vision. However, some cases have shown poor visual recovery even after complete removal of the ERM. Since the introduction of spectral domain optical coherence tomography (SD-OCT) was demonstrated to be useful for the noninvasive diagnosis and monitoring of macular diseases, the appearance of the photoreceptor layer on SD-OCT images has been found to be an important prognostic factor for visual recovery and postoperative best-corrected visual acuity after MIVS. Although deciding when to perform a vitrectomy can be difficult, it might be better to schedule vitrectomy surgery while the photoreceptor inner segment/outer segment (IS/OS) junction remains intact. If the preoperative IS/OS junction has already been disrupted, the surgery should be scheduled as early as possible to avoid further disturbances of visual improvement.

Power of proteomics in linking oxidative stress and female infertility.

Endometriosis, PCOS, and unexplained infertility are currently the most common diseases rendering large numbers of women infertile worldwide. Oxidative stress, due to its deleterious effects on proteins and nucleic acids, is postulated to be the one of the important mechanistic pathways in differential expression of proteins and in these diseases. The emerging field of proteomics has allowed identification of proteins involved in cell cycle, as antioxidants, extracellular matrix (ECM), cytoskeleton, and their linkage to oxidative stress in female infertility related diseases. The aim of this paper is to assess the association of oxidative stress and protein expression in the reproductive microenvironments such as endometrial fluid, peritoneal fluid, and follicular fluid, as well as reproductive tissues and serum. The review also highlights the literature that proposes the use of the fertility related proteins as potential biomarkers for noninvasive and early diagnosis of the aforementioned diseases rather than utilizing the more invasive methods used currently. The review will highlight the power of proteomic profiles identified in infertility related disease conditions and their linkage with underlying oxidative stress. The power of proteomics will be reviewed with regard to eliciting molecular mechanisms for early detection and management of these infertility related conditions.

Non-invasive diagnosis of alcoholic liver disease.

Alcoholic liver disease (ALD) is the most common liver disease in the Western world. For many reasons, it is underestimated and underdiagnosed. An early diagnosis is absolutely essential since it (1) helps to identify patients at genetic risk for ALD; (2) can trigger efficient abstinence namely in non-addicted patients; and (3) initiate screening programs to prevent life-threatening complications such as bleeding from varices, spontaneous bacterial peritonitis or hepatocellular cancer. The two major end points of ALD are alcoholic liver cirrhosis and the rare and clinically-defined alcoholic hepatitis (AH). The prediction and early diagnosis of both entities is still insufficiently solved and usually relies on a combination of laboratory, clinical and imaging findings. It is not widely conceived that conventional screening tools for ALD such as ultrasound imaging or routine laboratory testing can easily overlook ca. 40% of manifest alcoholic liver cirrhosis. Non-invasive methods such as transient elastography (Fibroscan), acoustic radiation force impulse imaging or shear wave elastography have significantly improved the early diagnosis of alcoholic cirrhosis. Present algorithms allow either the exclusion or the exact definition of advanced fibrosis stages in ca. 95% of patients. The correct interpretation of liver stiffness requires a timely abdominal ultrasound and actual transaminase levels. Other non-invasive methods such as controlled attenuation parameter, serum levels of M30 or M65, susceptometry or breath tests are under current evaluation to assess the degree of steatosis, apoptosis and iron overload in these patients. Liver biopsy still remains an important option to rule out comorbidities and to confirm the prognosis namely for patients with AH.

Fractional Flow Reserve Derived from Coronary Imaging and Computational Fluid Dynamics.

The assessment of functional severity of atherosclerotic stenoses in patients with coronary artery disease by invasive fractional flow reserve (FFR) measurement requires coronary artery cannulation, advancement of a wire and intravenous adenosine infusion with inherent procedure-related risk and costs. Coronary computed tomographic angiography (CCTA) and rotational coronary angiography (RA) have been recently used in conjunction with computational fluid dynamics (CFD) and image-based modelling for the determination of FFR without the need for additional imaging, modification of acquisition protocols or administration of medication. FFR derived from CCTA was demonstrated as superior to measures of CCTA stenosis severity for determination of lesion-specific ischaemia. Estimation of FFR from RA images and CFD provides a less invasive alternative to conventional FFR measurement while estimated values are in agreement with measured values. These new, combined anatomic-functional assessments have the potential to simplify the noninvasive diagnosis of coronary artery disease with a single study to identify patients with ischaemia-causing stenosis who may benefit from revascularisation.

Nonalcoholic Fatty Liver Disease: Intravoxel Incoherent Motion Diffusion-weighted MR Imaging—An Experimental Study in a Rabbit Model

To evaluate the feasibility of using intravoxel incoherent motion (IVIM) diffusion-weighted imaging with multiple b values for the noninvasive diagnosis of nonalcoholic fatty liver disease (NAFLD). This study was approved by the institutional animal care and use committee. Twenty-seven 8-week-old rabbits were fed a variety of diets (from a standard diet to a high-fat, high-cholesterol diet) before IVIM diffusion-weighted imaging was performed with seven b values by using a 3-T magnetic resonance (MR) imaging unit. At histologic analysis of the animals, livers were categorized by NAFLD severity as normal, NAFLD, borderline nonalcoholic steatohepatitis (NASH), or NASH. The apparent diffusion coefficient and IVIM-derived parameters including true diffusion coefficient, pseudodiffusion coefficient, and perfusion fraction of the liver parenchyma were measured. Each parameter was correlated with NAFLD severity, and optimal cutoff values were determined by means of receiver operating characteristics analysis. Perfusion fraction was significantly lower in rabbits with NAFLD than in those with a normal liver, and it decreased further as severity of NAFLD increased, with medians of 22.2%, 14.8%, 11.3%, and 9.5% in the rabbits in the normal, NAFLD, borderline, and NASH groups, respectively (ρ = -0.83, P < .001). Apparent diffusion coefficient, true diffusion coefficient, and pseudodiffusion coefficient were not significantly different between the NAFLD severity groups. In terms of the diagnostic performance of perfusion fraction, area under the curve values were 0.984 (normal vs NAFLD or more severe disease), 0.959 (NAFLD or less severe vs borderline or more severe disease), and 0.903 (borderline or less severe vs NASH) with optimal cutoff values of 15.2%, 13.2%, and 11.0%, respectively. Perfusion fractions extracted from IVIM diffusion-weighted imaging may help in the differentiation of early stage NASH from simple steatosis.

Sensitive determination of endogenous hexanal and heptanal in urine by hollow-fiber liquid-phase microextraction prior to capillary electrophoresis with amperometric detection

Hexanal (Hex) and heptanal (Hep) in human blood have been regarded as potential biomarkers of lung cancer. In this work, a hollow-fiber liquid-phase microextraction (HF-LPME) method has been developed for the preconcentration of these trace aldehydes in urine samples. After derivatization with an electroactive compound 2-thiobarbituric acid, these two non-electroactive aldehydes were converted to electroactive adducts, therefore detectable by capillary zone electrophoresis with amperometric detection (CZE-AD) approach. Experimental conditions of derivatization, extraction, electrophoretic separation and detection were optimized. Under the optimum conditions, the enrichment factors for Hex and Hep could reach 320 and 355, respectively. The limits of detection for Hex and Hep were 2.7 and 0.97 nM, respectively; the average recoveries were in the range of 61-95% and relative standard deviation (RSD) values less than 8.5%. The present method has been applied to quantitative analysis of two biomarkers in human urine in lieu of blood samples, and the assay results showed that the contents of Hex (0.99-6.7 μM) and Hep (2.5-6.4 μM) found in the urine sample of the lung cancer patients were significantly higher than those in the healthy volunteers, liver cancer patients, as well as diabetics. The proposed HF-LPME/CZE-AD method may provide a potential alternative for early non-invasive diagnosis of lung cancer disease.

Nonalcoholic Fatty Liver Disease: Intravoxel Incoherent Motion Diffusion-weighted MR Imaging-An Experimental Study in a Rabbit Model.

Purpose To evaluate the feasibility of using intravoxel incoherent motion (IVIM) diffusion-weighted imaging with multiple b values for the noninvasive diagnosis of nonalcoholic fatty liver disease (NAFLD). Materials and Methods This study was approved by the institutional animal care and use committee. Twenty-seven 8-week-old rabbits were fed a variety of diets (from a standard diet to a high-fat, high-cholesterol diet) before IVIM diffusion-weighted imaging was performed with seven b values by using a 3-T magnetic resonance (MR) imaging unit. At histologic analysis of the animals, livers were categorized by NAFLD severity as normal, NAFLD, borderline nonalcoholic steatohepatitis (NASH), or NASH. The apparent diffusion coefficient and IVIM-derived parameters including true diffusion coefficient, pseudodiffusion coefficient, and perfusion fraction of the liver parenchyma were measured. Each parameter was correlated with NAFLD severity, and optimal cutoff values were determined by means of receiver operating characteristics analysis. Results Perfusion fraction was significantly lower in rabbits with NAFLD than in those with a normal liver, and it decreased further as severity of NAFLD increased, with medians of 22.2%, 14.8%, 11.3%, and 9.5% in the rabbits in the normal, NAFLD, borderline, and NASH groups, respectively (ρ = -0.83, P < .001). Apparent diffusion coefficient, true diffusion coefficient, and pseudodiffusion coefficient were not significantly different between the NAFLD severity groups. In terms of the diagnostic performance of perfusion fraction, area under the curve values were 0.984 (normal vs NAFLD or more severe disease), 0.959 (NAFLD or less severe vs borderline or more severe disease), and 0.903 (borderline or less severe vs NASH) with optimal cutoff values of 15.2%, 13.2%, and 11.0%, respectively. Conclusion Perfusion fractions extracted from IVIM diffusion-weighted imaging may help in the differentiation of early stage NASH from simple steatosis. © RSNA, 2013.

Diagnosing coronary artery disease with hybrid PET/CT: It takes two to tango

The noninvasive diagnosis of coronary artery disease (CAD) is a challenging task. Although a large armamentarium of imaging modalities is available to evaluate the functional consequences of the extent and severity of CAD, cardiac perfusion positron emission tomography (PET) is considered the gold standard for this purpose. Alternatively, noninvasive anatomical imaging of coronary atherosclerosis with coronary computed tomography angiography (CCTA) has recently been successfully implemented in clinical practice. Although each of these diagnostic approaches has its own merits and caveats, functional and morphological imaging techniques provide fundamentally different insights into the disease process and should be considered to be complementary rather than overlapping. Hybrid imaging with PET/CT offers the possibility to evaluate both aspects nearly simultaneously, and studies have demonstrated that such a comprehensive assessment results in superior diagnostic accuracy, better prognostication, and helps in guiding clinical patient management. The aim of this review is to discuss the value of stand-alone CCTA and PET in CAD, and to summarize the available data on the surplus value of hybrid PET/CT including its strengths and limitations.

Distinctive features of microsaccades in Alzheimer's disease and in mild cognitive impairment.

During visual fixation, the eyes are never completely still, but produce small involuntary movements, called "fixational eye movements," including microsaccades, drift, and tremor. In certain neurological disorders, attempted fixation results in abnormal fixational eye movements with distinctive characteristics. Thus, determining how normal fixation differs from pathological fixation has the potential to aid early and differential noninvasive diagnosis of neurological disease as well as the quantification of its progression and response to treatment. Here, we recorded the eye movements produced by patients with Alzheimer's disease, patients with mild cognitive impairment, and healthy age-matched individuals during attempted fixation. We found that microsaccade magnitudes, velocities, durations, and intersaccadic intervals were comparable in the three subject groups, but microsaccade direction differed in patients versus healthy subjects. Our results indicate that microsaccades are more prevalently oblique in patients with Alzheimer's disease or mild cognitive impairment than in healthy subjects. These findings extended to those microsaccades paired in square-wave jerks, supporting the hypothesis that microsaccades and square-wave jerks form a continuum, both in healthy subjects and in neurological patients.

Multiparametric magnetic resonance for the non-invasive diagnosis of liver disease

Background & AimsWith the increasing prevalence of liver disease worldwide, there is an urgent clinical need for reliable methods to diagnose and stage liver pathology. Liver biopsy, the current gold standard, is invasive and limited by sampling and observer dependent variability. In this study, we aimed to assess the diagnostic accuracy of a novel magnetic resonance protocol for liver tissue characterisation.MethodsWe conducted a prospective study comparing our magnetic resonance technique against liver biopsy. The individual components of the scanning protocol were T1 mapping, proton spectroscopy and T2⁎ mapping, which quantified liver fibrosis, steatosis and haemosiderosis, respectively. Unselected adult patients referred for liver biopsy as part of their routine care were recruited. Scans performed prior to liver biopsy were analysed by physicians blinded to the histology results. The associations between magnetic resonance and histology variables were assessed. Receiver-operating characteristic analyses were also carried out.ResultsPaired magnetic resonance and biopsy data were obtained in 79 patients. Magnetic resonance measures correlated strongly with histology (rs = 0.68 p <0.0001 for fibrosis; rs = 0.89 p <0.001 for steatosis; rs = −0.69 p <0.0001 for haemosiderosis). The area under the receiver operating characteristic curve was 0.94, 0.93, and 0.94 for the diagnosis of any degree of fibrosis, steatosis and haemosiderosis respectively.ConclusionThe novel scanning method described here provides high diagnostic accuracy for the assessment of liver fibrosis, steatosis and haemosiderosis and could potentially replace liver biopsy for many indications. This is the first demonstration of a non-invasive test to differentiate early stages of fibrosis from normal liver.

Analysis of the Exhalome: A Diagnostic Tool of the Future

Investigations on breath analysis have provided preliminary data on its potential in the noninvasive diagnosis of lung diseases. Although the conventional comparisons of exhaled breath in study populations (ie, diseased vs healthy) may help to identify patients with various lung diseases, we believe that the analysis of exhaled breath holds promise beyond this scenario. On the basis of preliminary findings, we hypothesize that breath analysis (1) could be applied not only to identify patients with lung disease but also to better phenotype healthy subjects at risk and patients with a particular disease, which is in-line with current efforts toward individualized medicine; (2) could be useful in estimating internal body time to determine the optimal time of drug administration, thereby maximizing drug activity and reducing toxicity (chronopharmacology); and (3) could be applied to monitor drugs or drug metabolites, thus, enhancing adherence to prescribed medications and enabling studies on pharmacokinetics.

Stress echocardiography: safety and tolerability

BackgroundStress echocardiography is a valuable tool for the noninvasive diagnosis of ischemic heart disease. Despite its widely use in the clinical practice, safety and side effects profile have never been evaluated in Moroccans.The aimTo assess the safety and tolerability of the two stress echo modalities in Moroccans.MethodsThe study was made by 311 patients with known or suspected coronary artery disease, 203 underwent exercise echocardiography and 108 underwent dobutamine echocardiography, major and minor rhythmic complications and side effects were recorded for the two groups.ResultsWe registered 3 (2, 8%) major rhythmic events in the dobutamine group (2 sustained supraventricular tachycardia and 1 sustained ventricular tachycardia), there was no major rhythmic events in the exercise group. Minor rhythmic events were frequent (43, 5% in the dobutamine group and 19, 2% in the exercise group with a p = 0, 0001). Severe hypotension occurs in 4 (3, 7%) patients during a dobutamine stress, there was no significant drop in the blood pressure during exercise stress procedures. Non cardiac side effects were more common among patients who underwent a dobutamine stress echo (13, 9% vs. 3, 4% with p = 0,001).ConclusionExercise is safer than dobutamine stress echocardiography, complications and adverse effects with the use of dobutamine are usually minor and self-limiting.

Non-invasive prenatal diagnosis of genetic diseases by advanced technologies

The performance of three point of care methods for pleural fluid pH analysis was compared to a currently validated blood gas analyzer.An ABL 725 (Radiometer America, Westlake, OH) was used as the reference method to evaluate three cartridge-based assays: ABL 90 FLEX (Radiometer), and i-STAT 1 (Abbott Point of Care, Abbott Park, IL) CG4 + and G3 + cartridges for pleural fluid pH analysis. Pooled residual pleural fluid samples and quality control material were analyzed to determine intra- and inter-assay precision. Method comparison was performed with spiked (n = 40) and clinically-ordered (n = 10) pleural fluid samples across the analytical measuring range.All methods demonstrated inter-assay CVs < 0.1% at pH values of 7.1 and 7.6, and intra-assay CVs < 0.3% at pH values of 7.2 and 7.7. Bland–Altman plots demonstrated clinically significant bias between ABL 725 and each cartridge-based method only at pH > 7.6. For samples with pH < 7.6 mean bias vs. ABL 725 was − 0.01 for ABL 90 FLEX and 0.03 for i-STAT 1 CG4 + and G3 + cartridges. Clinical concordance using a decision limit of pH 7.2 was 96–98% for the three methods.Analytical and clinical performance of the three cartridge-based methods was comparable to a validated blood gas analyzer for pleural fluid pH analysis. Cartridge-based pH methods offer the advantage of easier troubleshooting for clots and clogs as they use disposable electrodes. However cartridge-based methods are not currently FDA-approved for pleural fluid samples, such that additional validation would be required for this specimen type.

Feasibility and reproducibility of spleen transient elastography and its role in combination with liver transient elastography for predicting the severity of chronic viral hepatitis

Liver transient elastography (L-TE) is a reliable, noninvasive predictor of disease severity in chronic liver disease of viral aetiology (CLD). Owing to the relationships among severity of CLD, portal hypertension and spleen involvement, the assessment of splenic stiffness (S-TE) may have an added value in staging CLD. Of 132 CLD patients of viral aetiology, 48 with myeloproliferative disorders (MD) and 64 healthy volunteers (HV), were concurrently investigated by both L-TE and S-TE. Liver disease severity was staged by liver biopsy (LB; Metavir) taken concurrently with TE examination and upper gastrointestinal tract endoscopy for gastro-oesophageal varices. The S-TE inter-observer agreement was analysed by an intra-class correlation coefficient (ICC); L-TE and S-TE accuracy was evaluated by receiver operating characteristic (ROC) curve analysis. Logistic regression analysis assessed the independent effect of L-TE and S-TE as predictors of hepatic fibrosis stage. S-TE failed in 22 CLD (16.6%), 12 (25%) MD and 12 (18%) HV. In the three groups, the ICC was 0.89 (0.84-0.92), 0.90 (0.85-0.94) and 0.86(0.80-0.91), respectively. In the CLD group, L-TE and S-TE independently predicted significant fibrosis (OR 5.2 and 4.6) and cirrhosis (OR 7.8 and 9.1), but at variance from L-TE, S-TE was independent from liver necroinflammation and steatosis. The NPV of S-TE for gastro-oesophageal varices was 100% using a 48 kPa cut-off. In CLD, spleen stiffness alone or in combination with hepatic stiffness can be reliably and reproducibly assessed by TE with the added value of improving the noninvasive diagnosis of severe liver disease and excluding the presence of oesophageal varices.