Standard Equipment Research Articles

Quantifying the reversibility of ATP hydrolysis in rabbit skeletal myosin subfragment 1 using a luciferase assay

BackgroundIt is known that the adenosine triphosphate (ATP) hydrolysis in a number of ATPases is reversible. Radioactive methods are standard in measuring ATPase reversibility. We used myosin as a well-studied model enzyme to develop a luciferase-based assay to quantify the reversibility of ATP hydrolysis. In myosin, ATP bound to the active site is hydrolyzed into adenosine diphosphate (ADP) and inorganic phosphate (Pi) and recombined back into ATP multiple times before products of hydrolysis are released. The reversibility of ATP hydrolysis by skeletal myosin was previously confirmed using radioactively labeled isotopes. Transient kinetics studies indicated that the ATP hydrolysis step is temperature-sensitive, with a dissociation equilibrium constant of 1.6 at 3 °C. Consequently, the association equilibrium constant at this temperature is 0.63. The goal of our work was two-fold, (a) to develop a luciferase-based assay to measure the equilibrium constant of enzymatic ATP hydrolysis, eliminating the need for radioactively labeled Pi, and (b) refine the value of the association equilibrium constant of the myosin ATPase hydrolysis step.MethodsIn this assay, a reaction mixture containing myosin and saturating levels of ADP and Pi was incubated to reach equilibrium, then the reaction was terminated, and the amount of ATP produced by myosin was quantified using the luciferase assay. The equilibrium constant of ATP hydrolysis was defined as the ratio of ATP to ADP bound to myosin.ResultsWe obtained a value of 0.78 ± 0.14 for the association equilibrium constant of ATP hydrolysis at 0 °C. 50 μM ADP bound to myosin is turned into 21.9 ± 3.0 μM ATP. Our result is in excellent agreement with the literature data, supporting the viability of the new methodology.DiscussionThis methodology allows a more accessible and safe option to measure ATP production and reversibility of ATPase enzymes. Standard laboratory equipment is utilized in the assay, and the number of steps in the developed assay is reduced significantly compared to the radioactive method.

Hiking Backpack Design Development with Integrated Splint Feature for Fracture Injuries

The background of this design is the risk of fracture injury to climbers when mountaineering. According to the International Alpine Trauma Registry (IATR), there were 306 mountaineering accidents resulting in fractures in 2019. First aid for fractures requires standardized equipment and methods to assist the rescue process. The problem studied is the difficulty of climbers in utilizing climbing equipment as splints when treating fracture victims on the mountain. The purpose of this design is to develop climbing equipment so that it can be converted into a means of bandaging to perform first aid for fracture injuries. The design method used is Quality Function Deployment (QFD), to determine the priority aspects related to the specifications of the means of massaging that users want. The result of the design is a backpack product with an internal frame that applies a collapsible mechanism so that it can be quickly accessed in an emergency and can be adjusted according to the dimensions of the body part that has a fracture injury. The developed splint design successfully meets the criteria for dimensional customization to the user's body and can immobilize the fracture according to standard first aid procedures.

Synthesis and Characterization of Lauha Bhasma

The preparation of ayurvedic bhasma is particularly challenging, as mass production in modern facilities often lacks the standardized equipment needed to ensure consistent quality, leading to concerns about toxic elements like mercury and arsenic in the final products. Addressing these concerns necessitates the standardization of the synthesis process through the implementation of stringent quality control measures at every stage of preparation. This study is dedicated to the synthesis and comprehensive characterization of lauha (iron) bhasma, employing both traditional methodologies and advanced analytical techniques. The synthesis process began with samanya shodhana (general purification), followed by vishesh shodhana (special purification) and marana (incineration) The resultant product then underwent amrutikaran, involving heating with aloe vera extract at an elevated temperature of 750°C.The synthesized lauha bhasma was meticulously characterized using a combination of classical Ayurvedic techniques and modern analytical methods, including Fourier Transform Infrared Spectroscopy (FTIR), Atomic Absorption Spectroscopy (AAS), X-ray Diffraction (XRD), Brunauer-Emmett-Teller (BET) surface analysis, Scanning Electron Microscopy (SEM), Particle Size Distribution (PSD), and Energy Dispersive X-ray (EDX) analysis. The final product, post-Amrutikaran, successfully passed all traditional Ayurvedic evaluations, affirming the proper formation of bhasma. XRD analysis revealed the formation of rhombohedral α-Fe₂O₃ with only trace amounts of free iron, indicating high purity. SEM and PSD analyses demonstrated the creation of nanosized particles ranging from 50 to 200 nm, complemented by a specific surface area of 12.55 m²/g as determined by BET studies. Crucially, EDX analysis confirmed that the amrutikaran process effectively eliminated mercury from the bhasma, ensuring its safety and compliance with stringent quality standards.

Parametry akustyczne ochronników słuchu i metody ich pomiaru

Acoustic parameters of hearing protectors and methods of their measurement The article presents how the basic parameter of hearing protectors, i.e. sound attenuation, is determined using a method based on the active participation of people (subjective method). In addition, objective methods for measuring acoustic parameters of hearing protectors, such as insertion loss, based on the use of miniature microphones and acoustic test fixture are described. The results of measurements of sound pressure level under hearing protectors carried out using an acoustic test fixture reproducing anatomical shape of a human head (including concha and ear canal), used as a standard for measuring acoustic parameters of hearing protectors in impulse noise situations are also presented. These measurements were carried out to verify the possibility of using this acoustic test fixture, to measure the sound pressure level reaching under the hearing protectors in the case of continuous noise. The tests showed that in the case of measurements with microphones that are standard equipment of the acoustic test fixture, dedicated to impulse noise, the use of these microphones is limited to high-level continuous signals. Microphones that are optional equipment on the acoustic test fixture were a more suitable solution for the intended purpose.

Advanced three-dimensional X-ray imaging unravels structural development of the human thymus compartments

BackgroundThe thymus, responsible for T cell-mediated adaptive immune system, has a structural and functional complexity that is not yet fully understood. Until now, thymic anatomy has been studied using histological thin sections or confocal microscopy 3D reconstruction, necessarily for limited volumes.MethodsWe used Phase Contrast X-Ray Computed Tomography to address the lack of whole-organ volumetric information on the microarchitecture of its structural components. We scanned 15 human thymi (9 foetal and 6 postnatal) with synchrotron radiation, and repeated scans using a conventional laboratory x-ray system. We used histology, immunofluorescence and flow cytometry to validate the x-ray findings.ResultsApplication to human thymi at pre- and post-natal stages allowed reliable tracking and quantification of the evolution of parameters such as size and distribution of Hassall’s Bodies and medulla-to-cortex ratio, whose changes reflect adaptation of thymic activity. We show that Hassall’s bodies can occupy 25% of the medulla volume, indicating they should be considered a third thymic compartment with possible implications on their role. Moreover, we demonstrate compatible results can be obtained with standard laboratory-based x-ray equipment, making this research tool accessible to a wider community.ConclusionsOur study allows overcoming the resolution and/or volumetric limitations of existing approaches for the study of thymic disfunction in congenital and acquired disorders affecting the adaptive immune system.

Inadequate Postmortem Reports: Investigating the Underlying Factors

Postmortem reports play a vital role in forensic pathology, providing essential information about the causes of death needed for legal investigations. However, several factors undermine the quality of these reports, threatening their reliability and compromising the integrity of the judicial system. This study explores the key issues contributing to inadequate postmortem reports, such as the absence of standardized protocols, lack of adequate training for forensic staff, communication breakdowns, external pressures on forensic teams, overwhelmed laboratories with insufficient infrastructure, personnel, and equipment, and challenges in implementing new technologies. By examining these elements, the research seeks to identify opportunities for improvement to enhance the quality and accuracy of autopsies. An integrated approach is crucial for establishing standardized practices, upgrading facilities and equipment, increasing staffing and laboratory numbers, improving training for forensic professionals, and promoting better communication among all stakeholders in forensic investigations. The findings emphasize the need to address these systemic issues to produce more reliable postmortem reports, which are essential for delivering accurate forensic insights and ensuring just legal outcomes.

Silage harvesting for small farms by using vacuum sealing in flexible polymer containers on a converted trailer

The article presents an analysis of milk production in Kazakhstan and identifies the reason for its low level, which is due to deficient feed, especially in small farms and in the private sector. The difficulty of saving the limited silage volume is due to the lack of preparation technology for preventing spoilage. The objective of this work is to complete the necessary equipment of a mobile tractor-trailer to reduce the specific energy consumption when preparing silage in flexible containers using a vacuum seal to increase the productivity of dairy cattle farming in smallholder and the private sector of the Republic. The basic rational parameters for sealing the silage by vacuum in the field on a mobile tractor-trailer for ease of transport and storage are obtained: silage weight in a flexible container – 769.6 kg, geometric dimensions of the sealed container by height (0.90 m), width (0.85 m) and length (0.85 m). The efficiency of the specific energy consumption of the proposed method for silage preparation is established at 35% compared with the traditional method. The recommended technology of silage preparation in flexible containers will be possible when conventional tractor-trailers are retrofitted with standard portable equipment (internal combustion engine-based (ICE) electric generator, vacuum pump, film welder).

A Holistic View of Asphalt Binder Aging under Ultraviolet Conditions: Chemical, Structural, and Rheological Characterization

Asphalt, as a key binder material in road construction, is susceptible to ultraviolet (UV) radiation-induced aging, leading to embrittlement and reduced durability. Despite the significance of UV aging, research in this area remains limited compared to that on thermal aging. This paper comprehensively reviews the current state of research on UV aging in asphalt, focusing on its mechanism, evaluation indicators, and methods to delay or avoid UV aging. The structural components, rheological properties, and aging mechanisms of asphalt are discussed. Various UV aging simulation methods, including the use of UV chambers and accelerated aging tests, are presented along with their evaluation tests such as dynamic shear rheometry, rutting tests, Fourier infrared spectroscopy, and bending beam rheology. Key indicators used to assess UV aging, including physical properties, rheological parameters, and chemical composition changes, are summarized. The mechanisms underlying UV aging, particularly the changes in asphalt’s structural components and rheological properties, are examined. The impact of factors like radiation intensity, temperature, chemical composition, and asphalt film thickness on UV aging is discussed. Additionally, various additives and modifiers, including modified bitumen, UV shielding agents, UV absorbers, antioxidants, and nanomodifiers, are reviewed for their potential to mitigate UV aging. This paper concludes by highlighting the challenges in developing standardized test equipment and evaluation criteria, the limitations of organic modifiers, and the need for further research on nanomaterials to improve asphalt’s UV aging resistance.

Analysis of grease mechanical degradation in standard equipment

As a grease is used in a bearing, its rheological properties may change substantially due to prolonged shear, causing it to no longer meet its application requirements. The change to grease structural integrity may be succinctly quantified by measuring the change in consistency, corresponding to shear/mechanical stability. There are only two standardized methods for measuring the mechanical stability of greases: prolonged working in a grease worker and subjecting to roller degradation in a roll stability test. This paper provides a detailed analysis of the shear environment of these two tests and uses recently identified measurement techniques to track the degradation of two different greases over time in each test. Results are then compared to shearing in a rheometer at shear rates that are estimated to be similar. It is shown that the shearing environments within the grease worker and roll stability test can be predicted and reproduced with reasonable accuracy in a rheometer. Various time scales of mechanical stability are identified, with the standard two-hour duration of the roll stability test found to exist within a transient time scale that may not accurately reflect the long-term mechanical stability of a grease. This study demonstrates that both tools can be used to reasonably estimate the changes to mechanical properties of a grease during the churning phase of a bearing or milling process during manufacturing, though it is recommended that the duration of the standard roll stability test be extended beyond the identified transient period.

Validation of the performance of a point of care molecular test for leprosy: From a simplified DNA extraction protocol to a portable qPCR.

The study aimed to optimize qPCR reactions using oligonucleotides from the first Brazilian molecular diagnostic kit for leprosy on a portable platform (Q3-Plus). In addition, we sought to develop a simplified protocol for DNA extraction that met point-of-care criteria. During optimization on the Q3-Plus, optical parameters, thresholds, and cutoffs for the 16S rRNA and RLEP targets of M. leprae were established using synthetic DNA, purified DNA from M. leprae, and pre-characterized clinical samples. For the simplified extraction protocol, different lysis solutions were evaluated using chaotropic agents, and purification was carried out by transferring the lysed material to FTA cards. The complete protocol (simplified extraction + qPCR on the portable platform) was then evaluated with pre-characterized clinical skin biopsy samples and compared with standard equipment (QuantStudio-5). LOD95% for the optimized reactions was 113.31 genome-equivalents/μL for 16S rRNA and 17.70 genome-equivalents/μL for RLEP. Among the lysis solutions, the best-performing was composed of urea (2 M), which provided good dissolution of the skin fragment and a lower Ct value, indicating higher concentrations of DNA. The complete technological solution showed a sensitivity of 52% in reactions. Our results highlight the need for additional optimization to deal with paucibacillary samples, but also demonstrate the feasibility of the portable platform for the qPCR detection of M. leprae DNA in low infrastructure settings.

Thermodynamic Calculations of the Main Characteristics of the Combustion Process of Pyrotechnic Nitrate-Metallized Mixtures with Additives of Organic and Inorganic Substances under External Thermal Influences

The aim of the article is to determine the dependencies of the combustion product temperatures of four-component mixtures, the content of high-temperature condensate, and unoxidized metal on the excess oxidizer coefficient, the number of additives, and external pressure using thermodynamic calculations. Modern methods of physicochemical analysis (such as cinephotography and microcinephotography methods, X-ray analysis methods, non-contact and contact temperature measurement methods), nonlinear thermal conductivity and thermal stability methods, as well as mathematical and experimental-statistical modeling, were employed to investigate the influence of elevated heating temperatures (up to 800 K) and external pressures (up to 107 Pa) on the ignition and combustion processes of pyrotechnic products. Standard pyrotechnic equipment was used to test pyrotechnic product samples under specified conditions. Calculations based on models were conducted in real-time and dialog mode on computer devices that meet modern requirements for the use of specialized software. The results of thermodynamic calculations are presented for the dependencies of combustion product temperatures of multicomponent pyrotechnic mixtures based on powders of metal fuels (magnesium, aluminium), nitrate oxidizer (sodium nitrate), additives of organic and inorganic substances (paraffin, sodium fluoride), and the content of high-temperature condensate and unoxidized metal depending on the excess oxidizer coefficient α = 0.1…4.0, the amount of paraffin additive εp = 0…0.2, sodium fluoride additive εf = 0…0.06, and external pressure P = 105…107 Pa. Based on the obtained results of thermodynamic calculations, statistical models in the form of regression dependencies of the specified combustion characteristics on their technological parameters and external conditions have been developed. These models enable the determination of fire hazardous properties of mixtures in real-time dialog and PC-based scenarios, considering premature initiation of products based on them under external thermal actions.

Unique polymer‐based composite coating with high anti‐corrosion characteristics and functional fillers

AbstractThe objective of this research is to develop a composite polymer coating with increased thermal conductivity and corrosion resistance for protecting structural materials in heat exchange apparatuses. Chemically pure compounds such as graphene, boron nitride, tyrosine, sodium hydroxide, zinc pyrophosphate, ethyl acetate, and epoxy resin with the addition of secondary polypropylene were utilized as raw materials. The obtained coatings were evaluated using an emission scanning microscope equipped with an energy‐dispersive spectroscopy (EDS) detector. Electrochemical activity was measured using standard equipment within a frequency range of 0.1–0.01 Hz. Thermal conductivity and corrosion resistance were measured at 0.24 W/m K and 9.70 Ω/cm2 when using only epoxy resin with the addition of secondary polypropylene. When employing a composite consisting of epoxy resin with the addition of secondary polypropylene, boron nitride, graphene, tyrosine, and zinc pyrophosphate, the thermal conductivity and corrosion resistance values were 1.65 W/m K and 11.10 Ω/cm2, respectively. Polymer coatings containing 30% composite demonstrated the highest thermal conductivity (1.65 W/m K) and corrosion resistance (11.20 Ω/cm2). The research findings can be utilized to create polymer coatings with enhanced thermal conductivity and corrosion resistance values. This polymer coatings could serve as structural materials for heat exchange devices.Highlights Corrosion resistance increase with complexing of composite coating. Thermal conductivity increase with complexing of composite coating. Use of epoxy resin with secondary polypropylene make thermal conductivity value.

POLICY DRIVERS OF CHINA'S INTEGRATED ENERGY SERVICES: A CURRENT STATUS REVIEW

Conventional energy production and consumption patterns seriously restrict global sustainable development. In the energy transformation process, integrated energy services play a key role in integrating clean energy and renewing energy consumption patterns. The European Union, the United States, and Japan are leading in developing policy frameworks and integrating integrated energy services. This paper compares and analyzes how the European Union, the United States, Japan, and China respond to the challenges of energy transformation through different energy policy systems. The energy policy systems of the European Union, the United States, and Japan are discussed for reference by China. The review reveals that (1) assigning tasks based on the energy transformation strength of each region is conducive to accelerating the speed of carbon emission reduction in each region; (2) continuous government support and targeted incentives are conducive to attracting market capital, thereby stimulating the advancement of energy technology; and (3) formulating energy efficiency standards for basic equipment can lay a good foundation for energy transformation.

Metric Stability of One Month Handgrip Maximal and Explosive Isometric Strength Measured by Classic and Impulse Contractions

Abstract The aim of this study is to determine the metric stability of the one-month handgrip test (HGT) in order to define the contractile characteristics of the biological variation of maximal isometric strength (F max) and maximal isometric rate of force development (RFD max) of the handgrip in two different testing regimes (classic and impulse). The study was conducted with a total of 16 participants (11 men and 5 women). Testing was performed using an isometric handgrip probe with a standardized test protocol and equipment sports medical solutions (SMS). The results of F max showed a low relative standard error of the mean (RS = 1.33 %), a high value of inter-class correlation (ICC = 0.996), and no statistically significant change in trend (p > 0.05) during the testing period. Therefore, can conclude that the HGT procedure in classic mode can be used as a stable parameter in a human subject sample. However, the RFD max results showed a low RS (2.13 %) and a high ICC value (0.996), but a statistically significant change of trend (p < 0.05) during the measurement period. The regression constant (RCO) trend was 42.629 N/s, which can be attributed to learning or to the adaptive effects of the test procedure, which triggered similar adaptation processes as the training. In general, it can be concluded that the handgrip can be used to sensitively measure the effects of different long-term health improvements, or the effects of different medical/health exercises, rehabilitation programs, effects of medication applications, or dietary supplements for F max. However, further research should be conducted for the RFD max considering the metric stability parameters.

Research on the calibration method of winding deformation tester

Abstract The focus of this paper is the calibration method and process for winding deformation tester, encompassing its working principle, metrological characteristics, calibration conditions and measurement methods. The calibration conditions are divided into environmental conditions and measurement standards and other equipments. The measurement methods are divided into indication error of scanning frequency and indication error of amplitude ratio. The key steps in the calibration process are elaborated, including the selection of standard equipments, error analysis, data processing. The method’s feasibility is verified through experiments.

Fluidic Interface for Surface-based DNA Origami Studies.

Traditionally, the use of DNA origami nanostructures (DONs) to study early cell signaling processes has been conducted using standard laboratory equipment with DONs typically utilized in solution. Surface-based technologies simplify the microscopic analysis of cells treated with DON agents by anchoring them to solid substrates, thus avoiding the complications of receptor-mediated endocytosis. A robust microfluidic platform for real-time monitoring and precise functionalization of surfaces with DONs was developed here. The combination of controlled flow conditions with an upright total internal reflection fluorescence microscope enabled the kinetic analysis of the immobilization of DONs on DNA-functionalized surfaces. The results revealed that DON morphology and binding tags influence the binding kinetics and that DON hybridization on surfaces is more effective in microfluidic devices with larger-than-standard dimensions, addressing the low diffusivity challenge of DONs. The platform enabled the decoration of DONs with protein-binding ligands and in situ investigation of ligand occupancy on DONs to produce high-quality bioactive surfaces. These surfaces were used to recruit and activate the epidermal growth factor receptor (EGFR) through clustering in the membranes of living cancer cells (MCF-7) using an antagonistic antibody (Panitumumab). The activation was quantified depending on the interligand distances of the EGFR-targeting antibody.

Aktivitas Pengelolaan Wisata Petualangan Canyoning Berbasis Resiko

Canyoning adventure tourism is a form of nature tourism activity that provides extreme and challenging experiences for tourists. However, the high level of risk in this activity requires a comprehensive risk management approach to ensure the safety and comfort of participants. This study aims to analyze risk-based canyoning tourism management activities in the Canyoning & Caving area, Bogor, Indonesia. This study uses a qualitative method with data collection techniques in the form of in-depth interviews with tour managers, canyoning guides, and tourists who have participated in the activity. In addition, direct observations were made of field conditions and facilities provided. The results of the study indicate that the implementation of effective risk management is very important to minimize the potential for accidents and negative impacts on the environment. Some of the main risks identified include unpredictable weather conditions, terrain difficulty, and lack of participant experience. The risk mitigation strategies implemented include intensive training for guides, the use of international standard safety equipment, and the preparation of clear emergency procedures. This study is expected to be a reference for other canyoning tourism managers in developing safe and sustainable adventure tourism in Indonesia.

Fiber Bragg Grating Pulse and Systolic Blood Pressure Measurement System Based on Mach-Zehnder Interferometer.

A fiber Bragg grating (FBG) pulse and systolic blood pressure (SBP) measurement system based on the edge-filtering method is proposed. The edge filter is the Mach-Zehnder interferometer (MZI) fabricated by two fiber couplers with a linear slope of 52.45 dBm/nm. The developed system consists of a broadband light source, an edge filter, fiber Bragg gratings (FBGs), a coarse wavelength-division multiplexer (CWDM), and signal-processing circuits based on a field-programmable gate array (FPGA). It can simultaneously measure pulse pulsations of the radial artery in the wrist at three positions: Cun, Guan and Chi. The SBP can be calculated based on the pulse transit time (PTT) principle. The measurement results compared to a standard blood pressure monitor showed the mean absolute error (MAE) and standard deviation (STD) of the SBP were 0.93 ± 3.13 mmHg. The system meets the requirements of the Association for the Advancement of Medical Instrumentation (AAMI) equipment standards. The proposed system can achieve continuous real-time measurement of pulse and SBP and has the advantages of fast detection speed, stable performance, and no compression sensation for subjects. The system has important application value in the fields of human health monitoring and medical device development.

Advanced mathematical modeling for preciseestimation of CT energy spectrum using a calibration phantom

PurposeThis research aims to develop an advanced mathematical model using a CT calibration phantom to accurately estimate the CT energy spectrum in clinical settings, enhancing imaging quality and patient dose management. MethodsData were collected from a CT scanner using a CT calibration phantom at various energy levels (80, 100, 120, and 135 kVp). The data was optimized to refine the energy spectrum model, followed by cross-validation with Monte Carlo simulations. ResultsThe developed model demonstrated high precision in estimating the CT energy spectrum at all tested energy levels, with R-squared values above 0.9738 and an R-squared value of 0.9829 at 100 kVp. The model also showed low Normalized Root Mean Square Deviation (NRMSD) ranging from 0.6698 % to 1.8745 %. The Mean Energy Difference (ΔE) between the estimated and simulated spectrum consistently remained under 0.01 keV. These results were comparable to recent studies, which reported higher NRMSD and ΔE. ConclusionsThis study presents a significantly improved model for estimating the CT energy spectrum, offering greater accuracy than existing models. Its strengths include high precision and the use of standard equipment and algorithmic values. While the current use of 13 plugs is adequate, incorporating plugs with varied densities could enhance accuracy. This model has potential for improving imaging quality and optimizing patient dosing in clinical applications. Future trends may include extending energy spectrum estimation to megavoltage domains and integrating technologies like EPID and MVCT for better dose distribution prediction in high-energy photon beam therapy.

ASPYRE-Lung: validation of a simple, fast, robust and novel method for multi-variant genomic analysis of actionable NSCLC variants in FFPE tissue.

Genomic variant testing of tumors is a critical gateway for patients to access the full potential of personalized oncology therapeutics. Current methods such as next-generation sequencing are costly and challenging to interpret, while PCR assays are limited in the number of variants they can cover. We developed ASPYRE® (Allele-Specific PYrophosphorolysis REaction) technology to address the urgent need for rapid, accessible and affordable diagnostics informing actionable genomic target variants of a given cancer. The targeted ASPYRE-Lung panel for non-small cell carcinoma covers 114 variants in 11 genes (ALK, BRAF, EGFR, ERBB2, KRAS, RET, ROS1, MET & NTRK1/2/3) to robustly inform clinical management. The assay detects single nucleotide variants, insertions, deletions, and gene fusions from tissue-derived DNA and RNA simultaneously. We tested the limit of detection, specificity, analytical accuracy and analytical precision of ASPYRE-Lung using FFPE lung tissue samples from patients with non-small cell lung carcinoma, variant-negative FFPE tissue from healthy donors, and FFPE-based contrived samples with controllable variant allele fractions. The sensitivity of ASPYRE-Lung was determined to be ≤ 3% variant allele fraction for single nucleotide variants and insertions or deletions, 100 copies for fusions, and 200 copies for MET exon 14 skipping. The specificity was 100% with no false positive results. The analytical accuracy test yielded no discordant calls between ASPYRE-Lung and expected results for clinical samples (via orthogonal testing) or contrived samples, and results were replicable across operators, reagent lots, runs, and real-time PCR instruments with a high degree of precision. The technology is simple and fast, requiring only four reagent transfer steps using standard laboratory equipment (PCR and qPCR instruments) with analysis via a cloud-based algorithm. The ASPYRE-Lung assay has the potential to be transformative in facilitating access to rapid, actionable molecular profiling of tissue for patients with non-small cell carcinoma.