Use Of Clinical Tests Research Articles

Cross-culturally modified University of Pennsylvania Smell Identification Test for a Turkish population.

The University of Pennsylvania Smell Identification Test (UPSIT) is a well-developed and popular olfactory test, which has been validated in various populations. However, there was only one study in a Turkish population, and this study indicated that the North American version of the UPSIT was not enough to evaluate the olfactory functions of Turkish population. So, we developed a cross-cultural adaptation of the UPSIT, the UPSIT-Turkish (UPSIT-T). Thus, the goal of this study was to investigate the applicability of the UPSIT-T in healthy Turkish subjects. The study included 51 healthy individuals who reported having normal olfactory function. The participants were administered the North American version of the UPSIT firstly, and then, after a day UPSIT-T was applied to the participants. The results of two smell tests were compared, and the applicability of UPSIT-T was evaluated. The mean (standard deviation) value for correctly identified odors was 27.2 ± 5.7 (range, 14-38) with the UPSIT application, whereas the mean (standard deviation) value for correctly identified odors was 35.9 ± 3.1 for UPSIT-T. There was a statistically significant increase in the scores of the participants when UPSIT-T was performed (p < 0.001). The identification rates of 10 test odorants were <80% for our study group, and 2 of 10 were <70% for the UPSIT-T. The UPSIT-T modification is an adequate olfactory test for clinical use in a Turkish population.

Availability and payer coverage of BRCA1/2 tests and gene panels.

Availability and payer coverage of BRCA1/2 tests and gene panels.

Real-Time, Quantitative Lighting-up Detection of Telomerase in Urines of Bladder Cancer Patients by AIEgens.

As a biomarker for early cancer diagnosis, telomerase are one of the promising targets for cancer therapeutics. Inspired by the fluorescent emission principle of aggregation-induced emission fluorogens, we creatively designed an AIE-based turn-on method to detect telomerase activity from cell extracts. A positively charged fluorogen (TPE-Z) is not fluorescent when freely diffused in solution. The fluorescence of TPE-Z is enhanced with the elongation of the DNA strand which could light up telomere elongation process. By exploitation of it, we can detect telomerase activity from different cell lines (E-J, HeLa, MCF-7, and HLF) with high sensitivity and specificity. Moreover, our method is successfully employed to demonstrate the applications in bladder cancer diagnosis (41 urine specimens from bladder cancer patients and 15 urine specimens from normal people are detected). The AIE-based method provides a simple one-pot technique for quantification and monitoring of the telomerase activity and shows great potential for future use in clinical tests.

Modeling Heterogeneous Bacterial Populations Exposed to Antibiotics: The Logistic-Dynamics Case.

In typical in vitro tests for clinical use or development of antibiotics, samples from a bacterial population are exposed to an antibiotic at various concentrations. The resulting data can then be used to build a mathematical model suitable for dosing regimen design or for further development. For bacterial populations that include resistant subpopulations-an issue that has reached alarming proportions-building such a model is challenging. In prior work, we developed a related modeling framework for such heterogeneous bacterial populations following linear dynamics when exposed to an antibiotic. We extend this framework to the case of logistic dynamics, common among strongly resistant bacterial strains. Explicit formulas are developed that can be easily used in parameter estimation and subsequent dosing regimen design under realistic pharmacokinetic conditions. A case study using experimental data from the effect of an antibiotic on a gram-negative bacterial population exemplifies the usefulness of the proposed approach.

Exploring the Use of Sonographic Evaluation of the Glenohumeral Labrum

Glenohumeral (GH) dislocations of the shoulder are prevalent in young male subjects and appear to be increasing in incidence. The classic diagnostic evaluation of labral injuries such as this involves the use of clinical testing as well as imaging modalities such as computed tomography, magnetic resonance imaging, and radiography, none of which provides a dynamic evaluation. This study was done to replicate sonographic evaluation of the glenoid labrum in a preclinical model and use the axillary scanning approach to specifically assess the integrity of the anterior inferior labrum. Using sonographic guidance, minimal amounts of differently colored dyes were injected into regions of the glenoid labrum. This replication was able to image only the posterior superior, posterior inferior, and anterior inferior aspects of the GH labrum. Bankart lesions were identified in two shoulders, confirmed on dissection. The presence of dyes in the target areas within and around the labrum was visually confirmed. These findings suggest that the concept of using an axillary scanning approach for detecting labral tears and Bankart lesions in patients with acute and chronic shoulder dislocation may be worthwhile to incorporate into a shoulder-scanning protocol.

Role of Physical Evaluation in the Early Identification of Axial Spondyloarthritis: A Research Proposal.

The aim of this study was to present a rationale to explore the use of clinical tests for the sacroiliac joints to detect early axial spondyloarthritis (SpA) and to suggest a protocol to validate these clinical tests. Based on the European Guidelines for Diagnosis and Treatments of Pelvic Girdle Pain, we propose a set of six clinical tests to identify the likely presence of inflammation in the sacroiliac joints associated with early axial SpA. As magnetic resonance imaging (MRI) is the current gold standard used to identify inflammation in the sacroiliac joints, the results of the proposed set of clinical tests are compared with those from the MRI examinations. We hypothesize that specific clinical tests, which combine pain provocation and functional tests, for assessing the sacroiliac joints will help to identify early active inflammation at the sacroiliac joints in axial SpA. If such tests prove to be sensitive and specific, they could add further value to the diagnostic classification criteria for axial SpA.

Validation of a next-generation-sequencing cancer panel for use in the clinical laboratory.

Next-generation sequencing allows for high-throughput processing and sensitive variant detection in multiple genes from small samples. For many diseases, including cancer, a comprehensive mutational profile of a targeted list of genes can be used to simultaneously inform patient care, establish eligibility for ongoing clinical trials, and further research. To validate a pan-cancer, next-generation-sequencing assay for use in the clinical laboratory. DNA was extracted from 68 clinical specimens (formalin-fixed, paraffin-embedded; fine-needle aspirates; peripheral blood; or bone marrow) and 5 normal controls. Sixty-four DNA samples (94%; 64 of 68) were successfully processed with the TruSeq Amplicon Cancer Panel (Illumina Inc, San Diego, California) and sequenced in 4 sequencing runs. The data were analyzed at 4 different filter settings for sequencing coverage and variant frequency cutoff. Libraries created from 40 specimens could be successfully sequenced in a single run and still yield sufficient coverage for robust data analysis of individual samples. Sensitivity for mutation detection down to 5% was demonstrated using dilutions of clinical specimens and control samples. The test was highly repeatable and reproducible and showed 100% concordance with clinically validated Sanger sequencing results. Comparison to an alternate next-generation sequencing technology was performed by also processing 9 of the specimens with the AmpliSeq Cancer Hotspot Panel (version 2; Life Technologies, Grand Island, New York). Thirty of the 31 (97%) TruSeq-detected variants covered by the designs of both panels were confirmed. A sensitive, high-throughput, pan-cancer mutation panel for sequencing of cancer hot-spot mutations in 42 genes was validated for routine use in clinical testing.

A PSA-guided approach for a better diagnosis of prostatic adenocarcinoma based on MALDI profiling and peptide identification

BackgroundProstate cancer (PCa) is the second cause of mortality in men worldwide. The prostate-specific antigen (PSA) test is routinely adopted in diagnosis; nevertheless more reliable biomarkers are continuously under investigation by monitoring the release of molecules into the bloodstream. The serum protein profiles appear to provide cancer-specific fingerprints that help to discriminate patients (especially with low PSA level) from controls, improving the performance of existing clinical tests. MethodsSamples from healthy controls and PCa patients with low (≤4ng/mL) and high PSA (>4ng/mL) levels were analyzed by MALDI profiling, and by a multi fractionation approach coupled to ESI-MS for peaks identification. ResultsMALDI profiling achieved to detect 10 and 14 changed peaks (p-value <0.05), respectively, in PCa patients with low and high PSA versus controls. In particular, a peak identified as C3f fragment, resulted overexpressed in low PSA PCa patients. ConclusionsPSA test, coupled to MALDI profiling, is able to detect changes, specifically related to PCa, in low molecular weight protein range. Furthermore, for the first time in prostate cancer research, the identification and quantification of the small peptide C3f appears to be relevant for the detection of false negatives, providing an additive diagnostic power to PSA (p<0.01) and suggesting its use in clinical tests.

Fragmentation of sperm DNA using the TUNEL method

ObjectivesTo establish the validity of the TUNEL assay in determining sperm DNA fragmentation, the relationship between the degree of fragmentation and the seminal parameters and the sample needed to conduct the test. Material and methodsWe used semen samples from healthy fertile men (n=33), patients who consulted for infertility with a prescription for the TUNEL assay (n=77) and patients with intracytoplasmic sperm injection failure (n=20), analyzed according to the 2010 WHO. The TUNEL/propidium iodide test was performed by flow cytometry, on baseline and post-swim-up samples. ResultsThe cutoff value for the TUNEL assay (ROC curves) was 26%, with a sensitivity and specificity of 85% and 89%, respectively. The pre-swim-up and post-swim-up medians of the results from the TUNEL assay showed no significant differences (17.0% vs. 12.9%, respectively). However, 39.1% of the samples showed a difference greater than 15 in absolute value between the results of the baseline and post-swim-up TUNEL assays. The linear correlation study of the morphology, mobility and vitality using the post-swim-up TUNEL assay showed a greater correlation than preselection, with significant results (r: −0.394, p<.0001; r: −0.461, p<.0001; r: −0.526, p<.0001). ConclusionsThe TUNEL assay is a valid test for clinical use. DNA fragmentation is a factor independent from traditional semen tests. We found a greater susceptibility to damage generated in the laboratory procedures in the samples with lower quality. The sample of choice for evaluating DNA fragmentation will depend on whether the clinician is treating a natural or assisted fertilization.

Accuracy of clinical tests in the diagnosis of anterior cruciate ligament injury: a systematic review.

BackgroundNumerous clinical tests are used in the diagnosis of anterior cruciate ligament (ACL) injury but their accuracy is unclear. The purpose of this study is to evaluate the diagnostic accuracy of clinical tests for the diagnosis of ACL injury.MethodsStudy Design: Systematic review. The review protocol was registered through PROSPERO (CRD42012002069).Electronic databases (PubMed, MEDLINE, EMBASE, CINAHL) were searched up to 19th of June 2013 to identify diagnostic studies comparing the accuracy of clinical tests for ACL injury to an acceptable reference standard (arthroscopy, arthrotomy, or MRI). Risk of bias was appraised using the QUADAS-2 checklist. Index test accuracy was evaluated using a descriptive analysis of paired likelihood ratios and displayed as forest plots.ResultsA total of 285 full-text articles were assessed for eligibility, from which 14 studies were included in this review. Included studies were deemed to be clinically and statistically heterogeneous, so a meta-analysis was not performed. Nine clinical tests from the history (popping sound at time of injury, giving way, effusion, pain, ability to continue activity) and four from physical examination (anterior draw test, Lachman’s test, prone Lachman’s test and pivot shift test) were investigated for diagnostic accuracy. Inspection of positive and negative likelihood ratios indicated that none of the individual tests provide useful diagnostic information in a clinical setting. Most studies were at risk of bias and reported imprecise estimates of diagnostic accuracy.ConclusionDespite being widely used and accepted in clinical practice, the results of individual history items or physical tests do not meaningfully change the probability of ACL injury. In contrast combinations of tests have higher diagnostic accuracy; however the most accurate combination of clinical tests remains an area for future research.Clinical relevanceClinicians should be aware of the limitations associated with the use of clinical tests for diagnosis of ACL injury.

Validation of Human Clinical Genetic Tests

In the last ten years, a high amount of genetic assays has been developed for molecular biopathology and genetic laboratories of the hospitals, mainly developed and provided by external companies. In some cases, the specialized staff members of the hospitals (doctors, biopathologists, geneticists or pharmacists) develop their own methods. The validation of these methods is required before their use in clinical testing, in order to assess its reliability. Analytical methods are validated under the requirements of International Guidelines, but validation procedures for clinical genetic tests are under study and need clarifications. In this manuscript, the main information related to the field of genetic validation is revised, including statistics, explaining the difficulty of validation for some of the developed genetic tests. The provided information is in agreement with all the International Guides. The information could be useful by the workers daily performing this kind of analysis.

LC-MS/MS determination of apixaban (BMS-562247) and its major metabolite in human plasma: an application of polarity switching and monolithic HPLC column.

apixaban (BMS-562247) (Eliquis(®)) is a novel, orally active, selective, direct, reversible inhibitor of the coagulation factor Xa (FXa). A sensitive and reliable method was developed and validated for the measurement of apixaban (BMS-562247) and its major circulating metabolite (BMS-730823) in human citrated plasma for use in clinical testing. A 0.100 ml portion of citrated plasma sample was extracted and analyzed by LC-MS/MS. Run times were approximately 3 min. The lower limit of quantification (LLOQ) was 1.00 ng/ml for BMS-562247 and 5.00 ng/ml for BMS-730823. Intra- and inter-assay precision values for replicate QC control samples were within ≤5.36% for both analytes (≤7.52% at the LLOQ). The accuracy for both analytes was within ±9.00%. The method was demonstrated to be sensitive, selective and robust, and was successfully used to support clinical studies.

Fragmentación del ADN espermático empleando el método de TUNEL

ObjectivesTo establish the validity of the TUNEL assay in determining sperm DNA fragmentation, the relationship between the degree of fragmentation and the seminal parameters and the sample needed to conduct the test. Material and methodsWe used semen samples from healthy fertile men (n=33), patients who consulted for infertility with a prescription for the TUNEL assay (n=77) and patients with intracytoplasmic sperm injection failure (n=20), analyzed according to the 2010 WHO. The TUNEL/propidium iodide test was performed by flow cytometry, on baseline and post-swim-up samples. ResultsThe cutoff value for the TUNEL assay (ROC curves) was 26%, with a sensitivity and specificity of 85% and 89%, respectively. The pre-swim-up and post-swim-up medians of the results from the TUNEL assay showed no significant differences (17.0% vs. 12.9%, respectively). However, 39.1% of the samples showed a difference greater than 15 in absolute value between the results of the baseline and post-swim-up TUNEL assays. The linear correlation study of the morphology, mobility and vitality using the post-swim-up TUNEL assay showed a greater correlation than preselection, with significant results (r: –0.394, P<.0001; r: –0.461, P<.0001; r: –0.526, P<.0001). ConclusionsThe TUNEL assay is a valid test for clinical use. DNA fragmentation is a factor independent from traditional semen tests. We found a greater susceptibility to damage generated in the laboratory procedures in the samples with lower quality. The sample of choice for evaluating DNA fragmentation will depend on whether the clinician is treating a natural or assisted fertilization.

690 The use of clinical tests for diagnosing detrusor underactivity in adult men – results of a pilot study

690 The use of clinical tests for diagnosing detrusor underactivity in adult men – results of a pilot study

Addressing the needs of traumatic brain injury with clinical proteomics

BackgroundNeurotrauma or injuries to the central nervous system (CNS) are a serious public health problem worldwide. Approximately 75% of all traumatic brain injuries (TBIs) are concussions or other mild TBI (mTBI) forms. Evaluation of concussion injury today is limited to an assessment of behavioral symptoms, often with delay and subject to motivation. Hence, there is an urgent need for an accurate chemical measure in biofluids to serve as a diagnostic tool for invisible brain wounds, to monitor severe patient trajectories, and to predict survival chances. Although a number of neurotrauma marker candidates have been reported, the broad spectrum of TBI limits the significance of small cohort studies. Specificity and sensitivity issues compound the development of a conclusive diagnostic assay, especially for concussion patients. Thus, the neurotrauma field currently has no diagnostic biofluid test in clinical use.ContentWe discuss the challenges of discovering new and validating identified neurotrauma marker candidates using proteomics-based strategies, including targeting, selection strategies and the application of mass spectrometry (MS) technologies and their potential impact to the neurotrauma field.SummaryMany studies use TBI marker candidates based on literature reports, yet progress in genomics and proteomics have started to provide neurotrauma protein profiles. Choosing meaningful marker candidates from such ‘long lists’ is still pending, as only few can be taken through the process of preclinical verification and large scale translational validation. Quantitative mass spectrometry targeting specific molecules rather than random sampling of the whole proteome, e.g., multiple reaction monitoring (MRM), offers an efficient and effective means to multiplex the measurement of several candidates in patient samples, thereby omitting the need for antibodies prior to clinical assay design. Sample preparation challenges specific to TBI are addressed. A tailored selection strategy combined with a multiplex screening approach is helping to arrive at diagnostically suitable candidates for clinical assay development. A surrogate marker test will be instrumental for critical decisions of TBI patient care and protection of concussion victims from repeated exposures that could result in lasting neurological deficits.

FDA‐approved Next‐Generation sequencing system could expand clinical genomic testing

FDA‐approved Next‐Generation sequencing system could expand clinical genomic testing

Review article: next generation diagnostic modalities in gastroenterology – gas phase volatile compound biomarker detection

The detection of airborne gas phase biomarkers that emanate from biological samples like urine, breath and faeces may herald a new age of non-invasive diagnostics. These biomarkers may reflect status in health and disease and can be detected by humans and other animals, to some extent, but far more consistently with instruments. The continued advancement in micro and nanotechnology has produced a range of compact and sophisticated gas analysis sensors and sensor systems, focussed primarily towards environmental and security applications. These instruments are now increasingly adapted for use in clinical testing and with the discovery of new gas volatile compound biomarkers, lead naturally to a new era of non-invasive diagnostics. To review current sensor instruments like the electronic nose (e-nose) and ion mobility spectroscopy (IMS), existing technology like gas chromatography-mass spectroscopy (GC-MS) and their application in the detection of gas phase volatile compound biomarkers in medicine - focussing on gastroenterology. A systematic search on Medline and Pubmed databases was performed to identify articles relevant to gas and volatile organic compounds. E-nose and IMS instruments achieve sensitivities and specificities ranging from 75 to 92% in differentiating between inflammatory bowel disease, bile acid diarrhoea and colon cancer from controls. For pulmonary disease, the sensitivities and specificities exceed 90% in differentiating between pulmonary malignancy, pneumonia and obstructive airways disease. These sensitivity levels also hold true for diabetes (92%) and bladder cancer (90%) when GC-MS is combined with an e-nose. The accurate reproducible sensing of volatile organic compounds (VOCs) using portable near-patient devices is a goal within reach for today's clinicians.

Accuracy of clinical tests in the diagnosis of anterior cruciate ligament injury: a systematic review

Numerous clinical tests are used in the diagnosis of anterior cruciate ligament (ACL) injury but their accuracy is unclear. The purpose of this study is to evaluate the diagnostic accuracy of clinical tests for the diagnosis of ACL injury. Study Design: Systematic review. The review protocol was registered through PROSPERO (CRD42012002069). Electronic databases (PubMed, MEDLINE, EMBASE, CINAHL) were searched up to 19th of June 2013 to identify diagnostic studies comparing the accuracy of clinical tests for ACL injury to an acceptable reference standard (arthroscopy, arthrotomy, or MRI). Risk of bias was appraised using the QUADAS-2 checklist. Index test accuracy was evaluated using a descriptive analysis of paired likelihood ratios and displayed as forest plots. A total of 285 full-text articles were assessed for eligibility, from which 14 studies were included in this review. Included studies were deemed to be clinically and statistically heterogeneous, so a meta-analysis was not performed. Nine clinical tests from the history (popping sound at time of injury, giving way, effusion, pain, ability to continue activity) and four from physical examination (anterior draw test, Lachman’s test, prone Lachman’s test and pivot shift test) were investigated for diagnostic accuracy. Inspection of positive and negative likelihood ratios indicated that none of the individual tests provide useful diagnostic information in a clinical setting. Most studies were at risk of bias and reported imprecise estimates of diagnostic accuracy. Despite being widely used and accepted in clinical practice, the results of individual history items or physical tests do not meaningfully change the probability of ACL injury. In contrast combinations of tests have higher diagnostic accuracy; however the most accurate combination of clinical tests remains an area for future research. Clinicians should be aware of the limitations associated with the use of clinical tests for diagnosis of ACL injury.

Przydatność testów klinicznych oceniających ból o charakterze korzeniowym i pseudokorzeniowym w rozpoznaniu rwy kulszowej – badania pilotażowe

&lt;b&gt;Cel pracy:&lt;/b&gt; Zróżnicowana i złożona etiologia bólu z okolicy lędźwiowo-krzyżowej z promieniowaniem do kończyny dolnej wzbudza ciągłe spory i kontrowersje wokół kwestii diagnozowania i leczenia tej grupy chorych. Pacjenci, u których występuje ból odcinka lędźwiowo--krzyżowego pleców z promieniowaniem do kończyny dolnej, są dzieleni na tych, u których dolegliwości mają charakter korzeniowy i pseudokorzeniowy. Brak jednoznacznych kryteriów różnicujących te dwa komponenty bólu utrudnia ustalenie pochodzenia tych objawów. Celem pracy było zbadanie, które testy diagnostyczne mogą być przydatne w ocenie lokalizacji bólu i udziału tkanek miękkich w patomechanizmie rwy kulszowej. &lt;br /&gt; &lt;b&gt;Materiał i metody:&lt;/b&gt; Badaniu poddano 50 chorych, u których stwierdzono objawy rwy kulszowej i przeprowadzono test Lasegue’a, skrzyżowany test Lasegue’a (xSLR), test Bragarda oraz pięć testów oceniających stawy krzyżowo-biodrowe (sacroiliac joints – SI): test Yeomana, test Patricka, test Menela, test wypchnięcia i test kolców. &lt;br /&gt; &lt;b&gt;Wyniki: &lt;/b&gt;Wyniki testu Lasegue'a i testu Bragarda wykazywały istotną korelację u pacjentów z przepukliną krążka. U wszystkich pacjentów ze zmianami degeneracyjnymi krążka międzykręgowego (disc bulging) wynik testu Bragarda był negatywny. Skrzyżowany objaw Lasegue’a był dodatni tylko u pacjentów z przepukliną jądra miażdżystego. U wszystkich pacjentów, u których dodatni był wynik skrzyżowanego testu Lasegue'a, także wyniki testu Bragarda były dodatnie. Ponadto u 98% badanych dodatni był przynajmniej jeden z pięciu testów oceniających staw krzyżowo-biodrowy, wynik dla czterech pozytywnych testów stwierdzono u 20% badanych. &lt;br /&gt; &lt;b&gt;Wnioski&lt;/b&gt;: Wyniki badania pozwalają stwierdzić, że przeprowadzanie jednocześnie testów Lasegue’a i Bragarda jest uzasadnione, ale niewystarczające do potwierdzenia korzeniowego charakteru bólu rwy kulszowej. Pacjenci, u których przewlekle występują objawy rwy kulszowej wiążące się z uwypukleniem krążka, powinni być również ocenieni pod kątem zajęcia stawów krzyżowo-biodrowych.

A Validated RT-PCR Based Gene Signature for Tolerance in Kidney Transplant Recipients

Long-life immunosuppression is an important factor that contributes to shorten the long-term survival of kidney transplant recipients. Identifying patients in whom donor-specific tolerance has developed would constitute a major advance in their care. Since this ability would allow the minimization or even withdrawal of immunosuppressive therapy in selected patients. We have previously identified a unique set of 14 biomarkers of transplantation tolerance (Sagoo et al JCI 2010). Aims: The GAMBIT study aims to translate these defined biomarkers of tolerance into clinically useful identities. Methods: To simplify the test for clinical use we performed best subset selection using samples from two independent kidney transplant recipient cohorts (Sagoo et al, and Newel et al JCI 2010). This was approached by fitting all possible models from size 1 to 14, and the optimal model was selected via cross-validation using the area under the curve as indicator of performance. One cohort was used for discovery, and the second for replication with microarray data. Platform transition from expression observed in microarray to quantitative RT-PCR has been performed in the GAMBIT study. Housekeeping gene of reference used was HPRT. Results: Best-subset selection suggested that the expression of 3 genes (PNOC, SH2DB1 and TLR5) provided the best and most stable performance. The 3 genes signature could be used to classify tolerant recipients with a specificity and sensitivity of 0.82 and 0.84 respectively in the replication cohort. Using qRT-PCR, the 3 genes signature we found a sensitivity of 0.83 and specificity of 0.98 (AUC=0.95) to predict tolerance, on a third independent cohort of 6 tolerant, 44 patients with stable function, and 13 chronic rejectors.Figure: [3 Genes ROC Curve]Conclusions: Tolerant recipients can be identified using peripheral blood gene expression of just 3 genes measured by RT-PCR with high specificity and sensitivity. Validation for routine clinical use of these biomarkers would bring to the fore the possibility of personalized medicine.