Use Of Clinical Tests Research Articles

Development and Analytical Validation of an Expanded Mutation Detection Panel for Next-Generation Sequencing of Thyroid Nodule Aspirates

Molecular analysis is used to evaluate the risk of malignancy for thyroid fine-needle aspirates, identified as indeterminate by microscopic cytology, on the basis of the detection of various oncogenic DNA mutations and fusion transcripts or on the use of various mRNAs or miRNA-based classifier algorithms. Our approach has been to use a combination test using the detection of oncogenic mutations/fusion transcripts and an miRNA expression-based classifier algorithm. To improve the performance of the combination test, the next-generation sequencing (NGS)-based mutational panel was expanded from the detection of 5 oncogenes to 10 oncogenes and tumor suppressor genes and the detection of fusion transcripts was increased from 6 to 38. Herein, we describe the assay development of the expanded panel NGS test and optimization of various steps for the library preparation of multiplexed target genes to maintain quality parameters for sequencing and to improve the robustness of the test for use in clinical testing in a College of American Pathologists/Clinical Laboratory Improvements Amendments-certified laboratory. Technical hurdles in NGS library preparation for the sequencing of both normal and high guanine-cytosine-rich regions, and balanced amplification of various amplicons in highly multiplexed PCRs, were successfully overcome. Analytical validation as a laboratory-developed test (ThyGeNEXT) included the demonstration of assay reproducibility, lower limit of detection, as well as other fundamental quality parameters.

VR/AR head-mounted display system based measurement and evaluation of dynamic visual acuity

This study evaluated the dynamic visual acuity of candidates by implementing a King–Devick (K-D) test chart in a virtual reality head-mounted display (VR HMD) and an augmented reality head-mounted display (AR HMD). Hard-copy KD (HCKD), VR HMD KD (VHKD), and AR HMD KD (AHKD) tests were conducted in 30 male and female candidates in the age of 10S and 20S and subjective symptom surveys were conducted. In the subjective symptom surveys, all except one of the VHKD questionnaire items showed subjective symptoms of less than 1 point. In the comparison between HCKD and VHKD, HCKD was measured more rapidly than VHKD in all tests. In the comparison between HCKD and AHKD, HCKD was measured more rapidly than AHKD in Tests 1, 2, and 3. In the comparison between VHKD and AHKD, AHKD was measured more rapidly than VHKD in Tests 1, 2, and 3. In the correlation analyses of test platforms, all platforms were correlated with each other, except for the correlation between HCKD and VHKD in Tests 1 and 2. There was no significant difference in the frequency of errors among Tests 1, 2, and 3 across test platforms.VHKD and AHKD, which require the body to be moved to read the chart, required longer measurement time than HCKD. In the measurements of each platform, AHKD was measured closer to HCKD than VHKD, which may be because the AHKD environment is closer to the actual environment than the VHKD environment. The effectiveness of VHKD and AHKD proposed in this research was evaluated experimentally. The results suggest that treatment and training could be performed concurrently through the use of clinical test and content development of VHKD and AHKD.

Immunophenotyping, plasticity tests and nanotagging of stem cells derived from adipose tissue of wild rodent agouti (Dasyprocta prymnolopha)

ABSTRACT There is a growing interest in the study of unspecialized mesenchymal stem cells, for there are still some discussions about their in vitro behavior. Regenerative medicine is a science undergoing improvement which develops treatments as cell therapy using somatic stem cells. In several studies, adipose tissue is presented as a source of multipotent adult cells that has several advantages over other tissue sources. This study aimed to characterize and evaluate the tagging of mesenchymal stem cells from the agoutis adipose tissue (Dasyprocta prymonolopha), with fluorescent intracytoplasmic nanocrystals. Fibroblast cells were observed, plastic adherent, with extended self-renewal, ability to form colonies, multipotency by differentiation into three lineages, population CD90 + and CD45 - expression, which issued high red fluorescence after the tagging with fluorescent nanocrystals by different paths and cryopreserved for future use. It is possible to conclude that mesenchymal stem cells from agouti adipose tissue have biological characteristics and in vitro behavior that demonstrate its potential for use in clinical tests.

LENGTH CHANGE OF THE ILIOFEMORAL LIGAMENT DURING TESTS FOR ANTERIOR MICROINSTABILITY OF THE HIP JOINT: A CADAVERIC VALIDITY STUDY.

There is little information to support the use of clinical tests to assess for microinstability of the hip joint. The purpose of this study was to use a string model to describe and compare length changes of the iliofemoral ligament in the test positions commonly used to assess hip ligament laxity. Twelve hip joints from nine cadavers (4 male; 5 female) with lifespans of 57-84 years of age were studied. A string model representing the medial and lateral arms of the iliofemoral ligament was secured to the proximal and distal attachment points. The amount of length change of the string model was compared in four test positions: 1) external rotation, 2) hyperextension-external rotation 3) abduction-extension-external rotation, and 4) adduction--extension-external rotation. For the medial arm, the greatest change occurred in the adduction-extension-external rotation position (12.7mm). This was significantly greater than the external rotation (5.1mm; p=0.002) and abduction-extension-external rotation position (1.9mm; p<0.001). The lateral arm also had the greatest excursion in the adduction-extension-external rotation position (16.6mm). This length change was significantly greater than the external rotation position (8.6mm; p=0.002), the hyperextension-external rotation (11.1mm; p=0.047), and the abduction-extension-external rotation position (5.6mm; p<0.001). Tests used for hip instability cause various levels of tension through the iliofemoral ligament. The combination of hip extension and external rotation increased the length change of string model and was maximized with hip adduction. The least amount of change occurred with the addition of hip abduction to extension and external rotation. Clinicians may use the information to help interpret tests for instability of the hip and may consider the combined position of hip extension, external rotation, and adduction to elucidate involvement of the iliofemoral femoral ligament. 2b; Exploratory cohort study with good reference standards.

SUN-068 Clinical Validation Of A Novel Cardio-metabolic Diagnostic Technology

Hypercholesterolemia is a cardio-metabolic disease caused by a disruption in low density lipoprotein (LDL) cholesterol metabolism. Proprotein convertase subtilisin-kexin 9 (PCSK9) is an indirect regulator of plasma LDL levels by controlling the number of LDL receptor molecules expressed at the plasma membrane. Serum levels of PCSK9, along with associated mutations, dictate the severity of the hypercholesterolemic condition. Although several treatment options are available to treat hypercholesterolemia, a cost-effective and quick test to best qualify patients for appropriate treatment is currently lacking. We have developed and validated a diagnostic tool to be used in a clinical setting which measures PCSK9 levels in a currently unreported manner. The results from this tool will give predictive instructions to physicians as to which hypercholesterolemic treatment will best suite the patient, preventing the development of alternative diseases (such as Type 1 and Type 2 diabetes) currently being observed by the trial-and-error methods of treatment. We have used disease (diabetic) and non-disease (non-diabetic) state human samples to validate the use of our diagnostic test for clinical use. The human samples were used in clinical trials to measure the effectiveness of the diagnostic tools in personalized medicine. The results showed the diagnostic tool has the ability to determine which patients have genetic mutations in PCSK9 that will result in the progression of their diabetic condition under generalized Statin recommendations.

Retronasal Olfaction Test Methods: A Systematic Review

Background:This report produces a bibliographic study of psychophysical tests proposed clinical assessments of retronasal olfaction.Aims:We review how these tests can be utilized and discuss their methodological properties.Study Design:Systematic review.Methods:We undertook a systematic literature review investigating the retronasal olfaction test methods. PubMed, the free online MEDLINE database on biomedical sciences, was searched for the period from 1984 to 2015 using the following relevant key phrases: “retronasal olfaction”, “orthonasal olfaction”, “olfaction disorders”, and “olfaction test”. For each of the selected titles cited in this study, the full manuscript was read and analyzed by each of the three authors of this paper independently before collaborative discussion for summation and analytical reporting. Two reviewers independently read the abstracts and full texts and categorised them into one of three subgroups as follow, suitable, not-suitable, and unsure. Then they cross-checked the results, and a third reviewer decided assigned the group “unsure” to either the suitable group or the not-suitable group. Fifty eight studies revealed as suitable for review by two authors whereas 13 found not suitable for review. The total amount of 60 uncertain (unsure) or differently categorized articles were further examined by the third author which resulted in 41 approvals and 19 rejections. Hence 99 approved articles passed the next step. Exclusion criteria were reviews, case reports, animal studies, and the articles of which methodology was a lack of olfaction tests. By this way excluded 69 papers, and finally, 30 original human research articles were taken as the data.Results:The study found that the three most widely used and accepted retronasal olfaction test methods are the retronasal olfaction test, the candy smell test and odorant presentation containers. All of the three psychophysical retronasal olfaction tests were combined with orthonasal tests in clinical use to examine and understand the smell function of the patient completely. There were two limitations concerning testing: “the lack concentrations and doses of test materials” and “performing measurements within the supra-threshold zone”.Conclusion:The appropriate test agents and optimal concentrations for the retronasal olfaction tests remain unclear and emerge as limitations of the retronasal olfaction test technique. The first step to overcoming these limitations will probably require identification of retronasal olfaction thresholds. Once these are determined, the concept of retronasal olfaction and its testing methods may be thoroughly reviewed.

Determination of indoles in plasma of polycystic ovarian syndrome patients by high performance liquid chromatography-fluorescence detection

A new method was established for the determination of indoles (indole (IND), 3-methylindole (3-MI), indolyl-3-acetic acid (IAA) and indolyl-3-propionic acid (IPA)) in plasma by high performance liquid chromatography-fluorescence detection (HPLC-FLD). The analytes were separated simultaneously on a Shim-pack VP-ODS column (150 mm×4.6 mm, 4.6 μm) using 15 mmol/L sodium dihydrogen phosphate solution and methanol (48:52, v/v) as the mobile phases. The column temperature was 30 ℃, and the flow rate was 0.8 mL/min. The calibration curves of IND, 3-MI, IPA and IAA showed good linear relationships. The intra-day and inter-day relative standard deviations (RSDs) for the analytes were both less than 6.31%. The average recoveries of the analytes in plasma were 97.5%-107.0%. The method was successfully applied to the analysis of indoles in the plasma of healthy women of reproductive age (n=25) as controls and patients with polycystic ovarian syndrome (n=61). The results showed that the concentrations of indoles in the plasma were significantly different between the two groups, and IND was found to be a risk factor and a potential diagnostic biomarker for polycystic ovarian syndrome (PCOS). The method is simple, sensitive and suitable for use in clinical testing and laboratory research.

OC73 SYSTEMATIC REVIEW OF THE PREDICTIVE ACCURACY OF POINT OF CARE TESTS OF COAGULOPATHY IN CARDIAC SURGERY

Abstract Background: Coagulopathic haemorrhage is a common and serious complication of cardiac surgery. Timely diagnosis and treatment of coagulopathy is considered an important determinant of outcome. The aims of the review were to quantify and compare the predictive accuracy of POC tests in clinical use. Methods: Observational studies and randomized controlled trials that report measures of predictive accuracy for POC tests that are, or have been used clinically for the diagnosis and treatment of coagulopathic haemorrhage. Results: 56 studies recruiting a total of 9123 participants, met the inclusion criteria for the qualitative analysis and 15 studies were included for quantitative analysis. We have shown that the existing evidence is of insufficient quality to determine the predictive accuracy of point of care diagnostic tests. The point estimates of the Summary AUCROC analyses and the analysis of sensitivity and specificity estimates from individual studies suggested that: Viscoelastic tests had low sensitivity and high specificity, Platelet Function Tests had low sensitivity and moderate specificity, and combined Viscoelastic and Platelet Function tests had high sensitivity and low specificity. However there was great uncertainty around all of these estimates and the 95% margins of uncertainty included values less 0.5 in every analysis. Calculated diagnostic odd ratios also indicated poor predictive accuracy for all of the groups. Conclusions: Coagulopathic haemorrhage remains a common and severe complication of cardiac surgery. A systematic review of available evidence did not demonstrate predictive accuracy for point-of-care tests in clinical use for the diagnosis of coagulopathy in cardiac surgery.

How to write an Interpretation

Every day we interpret examination findings and clinical tests with the aim of coming to a diagnosis. But how well do we interpret these tests? Whether it is a traditional...

Clinical assessment and patient-reported outcome measures in low-back pain – a survey among primary health care physiotherapists

Purpose: We aimed to map the physiotherapy practice in Sweden of clinical tests and patient-reported outcome measures in low-back pain (LBP), and to study advantages and barriers in using patient-reported outcome measures.Methods: An online survey was mailed to 4934 physiotherapists in primary health care in Sweden. Multiple choice questions investigated the use of clinical tests and patient-reported outcome measures in assessing patients with LBP. Open questions investigating the advantages and barriers to the use of patient-reported outcome measures were analyzed with content analysis.Results: The response rate was 25% (n = 1217). Clinical tests were used “always/often” by >60% of the participants, while most patient-reported outcome measures were used by <15%. Advantages in using patient-reported outcome measures were: the clinical reasoning process, to increase the quality of assessment, to get the patient’s voice, education and motivation of patients, and communication with health professionals. Barriers were lack of time and knowledge, administrative aspects, the interaction between physiotherapist and patient and, the applicability and validity of the patient-reported outcome measures.Conclusion: Our findings show that physiotherapists working in primary health care use clinical testing in LBP to a great extent, while various patient-reported outcome measures are used to a low-to-very-low extent. Several barriers to the use of patient-reported outcome measures were reported such as time, knowledge, and administrative issues, while important findings on advantages were to enhance the clinical reasoning process and to educate and motivate the patient. Barriers might be changed through education or organizational change-work. To enhance the use of patient-reported outcome measures and thus person-centered care in low-back pain, recommendation, and education on various patient-reported outcome measures need to be advocated.Implications for rehabilitationTo increase the effects of rehabilitation in low-back pain, yellow flags, and other factors need to be taken into the consideration in the assessment which means the use of patient-reported outcome measures in addition to clinical testing.The use of patient-reported outcome measures is an advantage in the clinical reasoning process to enhance the quality of assessment and to educate and motivate the patient.Barriers to use patient-reported outcome measures are mainly lack of time and knowledge, and administrative aspects.Through education or organizational change-work, barriers to the use of patient-reported outcome measures might be changed.

Preoperative Molecular Markers in Thyroid Nodules.

The need for distinguishing benign from malignant thyroid nodules has led to the pursuit of differentiating molecular markers. The most common molecular tests in clinical use are Afirma® Gene Expression Classifier (GEC) and Thyroseq® V2. Despite the rapidly developing field of molecular markers, several limitations exist. These challenges include the recent introduction of the histopathological diagnosis “Non-Invasive Follicular Thyroid neoplasm with Papillary-like nuclear features”, the correlation of genetic mutations within both benign and malignant pathologic diagnoses, the lack of follow-up of molecular marker negative nodules, and the cost-effectiveness of molecular markers. In this manuscript, we review the current published literature surrounding the diagnostic value of Afirma® GEC and Thyroseq® V2. Among Afirma® GEC studies, sensitivity (Se), specificity (Sp), positive predictive value (PPV), and negative predictive value (NPV) ranged from 75 to 100%, 5 to 53%, 13 to 100%, and 20 to 100%, respectively. Among Thyroseq® V2 studies, Se, Sp, PPV, and NPV ranged from 40 to 100%, 56 to 93%, 13 to 90%, and 48 to 97%, respectively. We also discuss current challenges to Afirma® GEC and Thyroseq® V2 utility and clinical application, and preview the future directions of these rapidly developing technologies.

Hepatic encephalopathy: current challenges and future prospects.

Hepatic encephalopathy (HE) is a common complication of liver dysfunction, including acute liver failure and liver cirrhosis. HE presents as a spectrum of neuropsychiatric symptoms ranging from subtle fluctuating cognitive impairment to coma. It is a significant contributor of morbidity in patients with liver disease. HE is observed in acute liver failure, liver bypass procedures, for example, shunt surgry and transjugular intrahepatic portosystemic shunt, and cirrhosis. These are classified as Type A, B and C HE, respectively. HE can also be classified according to whether its presence is overt or covert. The pathogenesis is linked with ammonia and glutamine production, and treatment is based on mechanisms to reduce the formation and/or removal of these compounds. There is no specific diagnostic test for HE, and diagnosis is based on clinical suspicion, excluding other causes and use of clinical tests that may support its diagnosis. Many tests are used in trials and experimentally, but have not yet gained universal acceptance. This review focuses on the definitions, pathogenesis and treatment of HE. Consideration will be given to existing treatment, including avoidance of precipitating factors and novel therapies such as prebiotics, probiotics, antibiotics, laxatives, branched-chain amino acids, shunt embolization and the importance of considering liver transplant in appropriate cases.

Fifty Years of Prefrontal Cortex Research: Impact on Assessment.

Our knowledge of the functions of the prefrontal cortex, often called executive, supervisory, or control, has been transformed over the past 50 years. After operationally defining terms for clarification, we review the impact of advances in functional, structural, and theoretical levels of understanding upon neuropsychological assessment practice as a means of identifying 11 principles/challenges relating to assessment of executive function. Three of these were already known 50 years ago, and 8 have been confirmed or emerged since. Key themes over this period have been the emergence of the use of naturalistic tests to address issues of "ecological validity"; discovery of the complexity of the frontal lobe control system; invention of new tests for clinical use; development of key theoretical frameworks that address the issue of the role of prefrontal cortex systems in the organization of human cognition; the move toward considering brain systems rather than brain regions; the advent of functional neuroimaging, and its emerging integration into clinical practice. Despite these huge advances, however, practicing neuropsychologists are still desperately in need of new ways of measuring executive function. We discuss pathways by which this might happen, including decoupling the two levels of explanation (information processing; brain structure) and integrating very recent technological advances into the neuropsychologist's toolbox. (JINS, 2017, 23, 755-767).

Clinical Assessment of Physical Examination Maneuvers for Superior Labral Anterior to Posterior Lesions.

Purpose Shoulder pain and disability pose a diagnostic challenge owing to the numerous etiologies and the potential for multiple disorders to exist simultaneously. The evidence to support the use of clinical tests for superior labral anterior to posterior complex (SLAP) is weak or absent. The purpose of this study is to determine the diagnostic validity of physical examination maneuvers for SLAP lesions by performing a methodologically rigorous, clinically applicable study. Methods We recruited consecutive new shoulder patients reporting pain and/or disability. The physician took a history and indicated their certainty about each possible diagnosis (“certain the diagnosis is absent/present,” or “uncertain requires further testing”). The clinician performed the physical tests for diagnoses where uncertainty remained. Magnetic resonance imaging arthrogram and arthroscopic examination were the gold standards. We calculated sensitivity, specificity, and likelihood ratios (LRs) and investigated whether combinations of the top tests provided stronger predictions. Results Ninety-three patients underwent physical examination for SLAP lesions. When using the presence of a SLAP lesion (Types I–V) as disease positive, none of the tests was sensitive (10.3–33.3) although they were moderately specific (61.3–92.6). When disease positive was defined as repaired SLAP lesion (including biceps tenodesis or tenotomy), the sensitivity (10.5–38.7) and specificity (70.6–93.8) of tests improved although not by a substantial amount. None of the tests was found to be clinically useful for predicting repairable SLAP lesions with all LRs close to one. The compression rotation test had the best LR for both definitions of disease (SLAP tear present = 1.8 and SLAP repaired = 1.67). There was no optimal combination of tests for diagnosing repairable SLAP lesions, with at least two tests positive providing the best combination of measurement properties (sensitivity 46.1% and specificity 64.7%). Conclusion Our study demonstrates that the physical examination tests for SLAP lesions are poor diagnostic indicators of disease. Performing a combination of tests will likely help, although the magnitude of the improvement is minimal. These authors caution clinicians placing confidence in the physical examination tests for SLAP lesions rather we suggest that clinicians rely on diagnostic imaging to confirm this diagnosis.

Assessing Genetic Expression Profiles in Melanoma Diagnosis.

Most melanocytic tumors can be characterized as a benign nevus or a melanoma by a trained pathologist using traditional histopathological methods. However, a minority demonstrates ambiguous features and continues to be a diagnostic challenge. Genetic expression profiling (GEP) assays have been developed in an effort to resolve this dilemma. These assays measure mRNA levels of specified genes using reverse transcription quantitative polymerase chain reaction technology. The development of GEP assays, methodology, challenges associated with GEP validation and testing, and the suitability of a currently available GEP test for clinical use are reviewed.

FDA-Cleared versus Laboratory-Developed Tests: Why Start from Scratch When Kits Are Available?

One of the most common questions facing laboratories when bringing up new clinical tests is deciding which specific assay best suits the needs of the lab, its patients, and its providers. At first glance, assays cleared by the Food and Drug Administration (FDA)2 for use in clinical testing would seem to have some obvious advantages: the validation requirements are less onerous than for laboratory-developed tests (LDTs) (1), and much of the legwork, e.g., sensitivity and specificity testing, is done by …

Reliability and validity of clinical tests to assess the anatomical integrity of the cervical spine in adults with neck pain and its associated disorders: Part 1-A systematic review from the Cervical Assessment and Diagnosis Research Evaluation (CADRE) Collaboration.

To determine the reliability and validity of clinical tests to assess the anatomical integrity of the cervical spine in adults with neck pain and its associated disorders. We updated the systematic review of the 2000-2010 Bone and Joint Decade Task Force on Neck Pain and its Associated Disorders. We also searched the literature to identify studies on the reliability and validity of Doppler velocimetry for the evaluation of cervical arteries. Two independent reviewers screened and critically appraised studies. We conducted a best evidence synthesis of low risk of bias studies and ranked the phases of investigations using the classification proposed by Sackett and Haynes. We screened 9022 articles and critically appraised 8 studies; all 8 studies had low risk of bias (three reliability and five validity Phase II-III studies). Preliminary evidence suggests that the extension-rotation test may be reliable and has adequate validity to rule out pain arising from facet joints. The evidence suggests variable reliability and preliminary validity for the evaluation of cervical radiculopathy including neurological examination (manual motor testing, dermatomal sensory testing, deep tendon reflexes, and pathological reflex testing), Spurling's and the upper limb neurodynamic tests. No evidence was found for doppler velocimetry. Little evidence exists to support the use of clinical tests to evaluate the anatomical integrity of the cervical spine in adults with neck pain and its associated disorders. We found preliminary evidence to support the use of the extension-rotation test, neurological examination, Spurling's and the upper limb neurodynamic tests.

Effects of 1,3,5-triphenyl-4,5-dihydro-1H-pyrazole derivatives on cell-cycle and apoptosis in human acute leukemia cell lines.

Pyrazoline is an important 5-membered nitrogen heterocycle that has been extensively researched. Ten derivatives were synthesized and tested for antileukemic effects on 2 human acute leukemia cell lines, K562 and Jurkat. The most cytotoxic of these derivatives, compound 21, was chosen for investigation of cytotoxicity mechanisms. The results obtained with selectivity calculations revealed that compound 21 is more selective for acute leukemia (K562 and Jurkat cell lines) than for other tumor cell lines. Moreover, compound 21 was not cytotoxic to normal cell lines, indicating a potential use in clinical tests. Compound 21 caused a significant cell cycle arrest in the S-phase in Jurkat cells and increased the proportion of cells in the sub G0/G1 phase in both cell lines. Cells treated with compound 21 demonstrated morphological changes characteristic of apoptosis in the EB/AO assay, confirmed by externalization of phosphatidylserine by the annexin V - fluorescein isothiocyanate method and by DNA fragmentation. An investigation of cytotoxicity mechanisms suggests the involvement of an intrinsic apoptosis pathway due to mitochondrial damage and an increase in the ratio of mitochondrial Bax/Bcl2. Pyrazoline 21 obeyed Lipinski's "rule of five" for drug-likeness. Based on these preliminary results, the antileukemic activity of compound 21 makes it a potential anticancer agent.

Recent Advances in Disease Modeling and Drug Discovery for Diabetes Mellitus Using Induced Pluripotent Stem Cells.

Diabetes mellitus (DM) is a widespread metabolic disease with a progressive incidence of morbidity and mortality worldwide. Despite extensive research, treatment options for diabetic patients remains limited. Although significant challenges remain, induced pluripotent stem cells (iPSCs) have the capacity to differentiate into any cell type, including insulin-secreting pancreatic β cells, highlighting its potential as a treatment option for DM. Several iPSC lines have recently been derived from both diabetic and healthy donors. Using different reprogramming techniques, iPSCs were differentiated into insulin-secreting pancreatic βcells. Furthermore, diabetes patient-derived iPSCs (DiPSCs) are increasingly being used as a platform to perform cell-based drug screening in order to develop DiPSC-based cell therapies against DM. Toxicity and teratogenicity assays based on iPSC-derived cells can also provide additional information on safety before advancing drugs to clinical trials. In this review, we summarize recent advances in the development of techniques for differentiation of iPSCs or DiPSCs into insulin-secreting pancreatic β cells, their applications in drug screening, and their role in complementing and replacing animal testing in clinical use. Advances in iPSC technologies will provide new knowledge needed to develop patient-specific iPSC-based diabetic therapies.

Label-free and Electrochemical Detection of Nucleic Acids Based on Isothermal Amplification in Combination with Solid-state pH Sensor

Label-free quantification methods for nucleic acids are attracting attention, since they provide potential tools in clinical applications such as disease diagnosis or prognosis observation. In this research, we developed a simpler and cost-effective electrical monitoring device of nucleic acid amplification, combining an isothermal amplification method and an Iridium/Iridium oxide (Ir/IrOx) electrode without labeling. The fabricated Ir/IrOx electrode showed ideal Nernstian response under the various pH buffer solutions, and its potential response was less affected even in the presence of charged proteins. Moreover, we successfully real-time monitored primer generation-rolling circle amplification (PG-RCA) at the 0, 10, 100, 1000 pM of target DNA by detection of released proton during amplification reaction at the constant reaction temperature. This label-free and portable device might be received considerable attention as a useful platform for a point-of-care testing in clinical use.