Good Repeatability Research Articles

Repeatability, interocular correlation and agreement of optic nerve head vessel density in healthy eyes: a swept-source optical coherence tomographic angiography study.

To assess the repeatability, interocular correlation, and agreement of quantitative swept-source optical coherence tomography angiography (OCTA) optic nerve head (ONH) parameters in healthy subjects. Thirty-three healthy subjects were enrolled. The ONH of both eyes were imaged four times by a swept-source-OCTA using a 3 mm ×3 mm scanning protocol. Images of the radial peripapillary capillary were analyzed by a customized Matlab program, and the vessel density, fractal dimension, and vessel diameter index were measured. The repeatability of the four scans was determined by the intraclass correlation coefficient (ICC). The most well-centered optic disc from the four repeated scans was then selected for the interocular correlation and agreement analysis using the Pearson correlation coefficient, ICC and Bland-Altman plots. All swept-source-OCTA ONH parameters exhibited certain repeatability, with ICC>0.760 and coefficient of variation (CoV)≤7.301%. The obvious interocular correlation was observed for papillary vessel density (ICC=0.857), vessel diameter index (ICC=0.857) and fractal dimension (ICC=0.906), while circumpapillary vessel density exhibited moderate interocular correlation (ICC=0.687). Bland-Altman plots revealed an agreement range of -5.26% to 6.21% for circumpapillary vessel density. OCTA ONH parameters demonstrate good repeatability in healthy subjects. The interocular correlations of papillary vessel density, fractal dimension and vessel diameter index are high, but the correlation for circumpapillary vessel density is moderate.

Suitability of multifunction devices Myah and Myopia Master for monitoring myopia progression in children and adults.

To assess the feasibility of using multifunction instruments to measure axial length for monitoring myopia progression in children and adults. Axial length was measured in 60 children (aged 6-18 years) and 60 adults (aged 19-50 years) with multifunction instruments (Myah and Myopia Master) and stand-alone biometers (Lenstar LS900 and IOLMaster 700). Repeatability (measurements by the same examiner) and reproducibility (measurements by different examiners) were computed as the within-subject standard deviation (Sw) and 95% limits of agreement (LoA). Inter-instrument agreement was computed as intraclass correlation coefficients. The threshold for detecting myopic progression was taken as 0.1 mm. Measures were repeated only in children following the administration of 1% tropicamide to determine the impact of cycloplegia on axial length. Overall, the IOLMaster 700 had the best repeatability in children (0.014 mm) and adults (0.009 mm). Repeatability Sw values for all devices ranged from 0.005 to 0.021 mm (children) and 0.003 to 0.016 mm (adults). In children, reproducibility fell within 0.1 mm 95% of the time for the Myah, Myopia Master and IOLMaster 700. Agreement among all devices was classified as excellent (ICC 0.999; 95% CI 0.998-0.999), but the 95% LoA among the Myah, Myopia Master and Lenstar LS900 was ≥0.1 mm. Cycloplegia had no statistically significant effect on axial length (all p > 0.13). The Myah and Myopia Master multifunction instruments demonstrated good repeatability and reproducibility, and their accuracy was comparable to stand-alone biometers. Axial length measurements using different instruments can be considered interchangeable but should be compared with some caution. Accurate axial length measurements can be obtained without cycloplegia. The multifunction instruments Myah and Myopia Master are as well suited for monitoring myopia progression in children asthe stand-alone biometers IOLMaster 700 and Lenstar LS900.

Detection method for reverse transcription recombinase-aided amplification of avian influenza virus subtypes H5, H7, and H9

BackgroundAvian influenza virus (AIV) not only causes huge economic losses to the poultry industry, but also threatens human health. Reverse transcription recombinase-aided amplification (RT-RAA) is a novel isothermal nucleic acid amplification technology. This study aimed to improve the detection efficiency of H5, H7, and H9 subtypes of AIV and detect the disease in time. This study established RT-RAA-LFD and real-time fluorescence RT-RAA (RF-RT-RAA) detection methods, which combined RT-RAA with lateral flow dipstick (LFD) and exo probe respectively, while primers and probes were designed based on the reaction principle of RT-RAA.ResultsThe results showed that RT-RAA-LFD could specifically amplify H5, H7, and H9 subtypes of AIV at 37 °C, 18 min, 39 °C, 20 min, and 38 °C, 18 min, respectively. The sensitivity of all three subtypes for RT-RAA-LFD was 102 copies/µL, which was 10 ∼100 times higher than that of reverse transcription polymerase chain reaction (RT-PCR) agarose electrophoresis method. RF-RT-RAA could specifically amplify H5, H7, and H9 subtypes of AIV at 40 °C, 20 min, 38 °C, 16 min, and 39 °C, 17 min, respectively. The sensitivity of all three subtypes for RF-RT-RAA was 101 copies/µL, which was consistent with the results of real-time fluorescence quantification RT-PCR, and 100 ∼1000 times higher than that of RT-PCR-agarose electrophoresis method. The total coincidence rate of the two methods and RT-PCR-agarose electrophoresis in the detection of clinical samples was higher than 95%.ConclusionsRT-RAA-LFD and RF-RT-RAA were successfully established in this experiment, with quick response, simple operation, strong specificity, high sensitivity, good repeatability, and stability. They are suitable for the early and rapid diagnosis of Avian influenza and they have positive significance for the prevention, control of the disease, and public health safety.

Electrochemical Detection of the Antidiabetic Medication Metformin in Human Urine and Pharmaceutical Tablets Using Boron-Doped Diamond Electrode

Electrochemical Detection of the Antidiabetic Medication Metformin in Human Urine and Pharmaceutical Tablets Using Boron-Doped Diamond Electrode

Rapid and Reliable Quantification of Urinary Malondialdehyde by HILIC-MS/MS: A Derivatization-free Breakthrough Approach

BackgroundThe development of fast analytical methods is crucial for the research, discovery, and confirmation of crucial biomarkers. Furthermore, the implementation of fast analytical strategies contributes to efficient and time-effective procedures. In this sense, analysis of malondialdehyde (MDA) has become an important tool for understanding the role of oxidative stress in various diseases and for evaluating the efficacy of therapeutic interventions. ResultsA rapid and robust liquid chromatography tandem mass spectrometry method (HPLC-MS/MS) has been developed to determine endogenous amounts of malondialdehyde (MDA) in human urine without any associated derivatization reaction. MDA was separated in 4 minutes through a Urea-HILIC column and was analyzed using a triple quadrupole mass spectrometer in negative electrospray ionization mode. With a 50-fold dilution as the only sample pretreatment after alkaline hydrolysis, no matrix effect was present, which allowed for a fast and simple quantification by means of an external standard calibration with a limit of detection of 0.20 ng mL-1. The whole methodology was validated by analyzing unspiked and spiked urine samples from ten healthy individuals and comparing with the results obtained by the standard addition method. MDA was detected in all cases, with natural concentrations varying from 0.11±0.03 to 0.31±0.03 mg g-1 creatinine. Accuracies were found to be satisfactory, ranging from 95% to 101%. The proposed method also exhibited good repeatability and reproducibility (RSD<15%) for four quality control levels. SignificanceThe main significance of this method is the avoidance of a derivatization reaction for the determination of urinary MDA, this constituting a step forward when compared with previous literature. This breakthrough not only streamlines time analysis to less than five minutes per sample but also results in a more robust procedure. Consequently, the method here developed could be applied to subsequent future research involving the determination of MDA as a lipid peroxidation biomarker, where simple, rapid, and reliable methods could represent a significant improvement.

Research on infrared spectroscopy detection of furfural content in transformer oil based on acetonitrile extraction

Abstract The stable operation of power transformers depends on the oil-paper insulation system, in which the aging degree of insulating paper is the key to evaluate the remaining life of the transformer. Currently, methanol extraction is widely used to detect the furfural content in oil to evaluate the aging state of insulating paper, but it ignores the influence of methanol produced during the operation of the transformer on the extraction results. To address this issue, this paper proposes a new method that can replace methanol extraction for detecting furfural in oil through simulation and experiment. Firstly, through simulation and experiment, it is proved that the strong vibration absorption peaks at 1677 cm-1 for furfural carbonyl and 2240 cm-1 for acetonitrile cyano can be used to establish a new method for extracting furfural content in oil using acetonitrile. A quantitative model between the concentration x of furfural in the furfural-acetonitrile mixed solution and the area y of the infrared absorption peak at 1677 cm-1 is established, with a goodness of fit of 0.9974. Secondly, a comparison between direct detection and acetonitrile extraction methods is conducted. The results show that direct detection is simple to operate, but the minimum detection concentration is 40mg/L, which is difficult to meet practical requirements. Acetonitrile extraction can reduce the minimum detection concentration to 0.1mg/L. At the same time, the extraction conditions are analyzed to determine the extraction ratio of 30, extraction times of 5, and extraction rate of 60%. Finally, the proposed detection method is applied to thermal aging tests on insulating paper of different types. The experimental results show that the proposed method has good repeatability and improves the detection resolution of furfural content. This paper combines infrared spectroscopy with acetonitrile extraction technology to open up a more efficient and practical new method for detecting furfural content in transformer oil.

Nucleic acid aptamer-based electrochemical sensor for the detection of serum P-tau231 and the instant screening test of Alzheimer's disease.

The instant screening of patients with a tendency towards developing Alzheimer's disease (AD) is significant for providing preventive measures and treatment. However, the current imaging-based technology cannot meet the requirements in the early stage. Developing biosensor-based liquid biopsy technology could be overcoming this bottleneck problem. Herein, we developed a simple, low-cost, and sensitive electrochemical aptamer biosensor for detecting phosphorylated tau protein threonine 231 (P-tau231), the earliest and one of the most efficacious abnormally elevated biomarkers of AD. Gold nanoparticles (AuNPs) were electrochemically synthesized on a glassy carbon electrode as the transducer, exhibiting excellent conductivity, and were applied to amplify the electrochemical signal. A nucleic acid aptamer was designed as the receptor to capture the P-tau231 protein, specifically through the formation of an aptamer-antigen complex. The proposed biosensor showed excellent sensitivity in detecting P-tau 231, with a broad linear detection range from 10 to 107pg/mL and a limit of detection (LOD) of 2.31pg/mL. The recoveries of the biosensor in human serum ranged from 97.59 to 103.26%, demonstrating that the biosensor could be used in complex practical samples. In addition, the results showed that the developed biosensor has good repeatability, reproducibility, and stability, which provides a novel method for the early screening of AD.

A Sensitive Electrochemical Aptamer Sensor for Clenbuterol Detection Based on Au NPs/Fe3O4–NH2/GN Nanocomposite

Chronic ingestion of food containing clenbuterol (CLB) in the human body can cause acute poisoning. Thus, it is essential to develop a sensitive CLB detection way. Here, a novel electrochemical aptamer sensor based on aptamer/gold nanoparticles/amino magnetic ferric oxide/graphene nanosheets (aptamer/Au NPs/Fe3O4–NH2/GN) nanocomposite has been developed. The Fe3O4–NH2 nanoparticles were self-assembled on GN, and then Fe3O4–NH2/GN nanocomposite was coated on the glassy carbon electrode (GCE) surface. Afterward, Au NPs were electrodeposited on Fe3O4–NH2/GN nanocomposite to fix the aptamer by the Au–S bonds. When CLB was added to the solution, the aptamer will specifically recognize CLB, causing current changes. Under the optimized conditions, the sensor shows better electrochemical performance (linear range is [Formula: see text][Formula: see text]M to [Formula: see text][Formula: see text]M, the detection limit is [Formula: see text][Formula: see text]M ([Formula: see text])). It has been successfully applied to the detection of CLB in actual samples. The results show that the aptamer sensor has good selectivity, stability and repeatability, which will provide a novel idea for more sensitive and quicker CLB detection.

Surface structural transformation and adsorption performance of Poly(triazine imide)

Surface structural transformation and adsorption performance of Poly(triazine imide)

Non-Enzymatic Glucose Detection by Fe2O3 Nanorods-Reduced Graphene under Physiological pH

Background: Non-enzymatic detection has become a research hotspot because of its alternativity in solving problems compared to enzymatic biosensors, but most of those sensors require a strong basic pH environment (higher than 10) to active their surface, restricting their use in clinical detection because the pH of body fluid is around 7.4. Furthermore, metal oxide sensors with specific morphologies are reported to have a fast electrocatalytic response. Therefore, Fe2O3 nanocomposites with porous structure are selected for glucose detection research in a physiological pH environment. Objective: The study aimed to assess the potential use of porous reduced graphene oxide-Fe2O3 nanorods in glucose detection in a physiological pH environment. Method: Hydrothermal method was used to prepare porous Fe2O3-rGO NRs (Nanorods) and hollow Fe2O3/C nanoparticles. Cyclic voltammetry and electrochemical impedance spectroscopy were used to evaluate the performance of our materials. Results: Porous-reduced graphene oxide-Fe2O3 nanorods have exhibited better performance than hollow carbon-Fe2O3 core-shell nanoparticles for glucose detection in a physiological pH environment. Conclusion: Non-enzymatic glucose sensing based upon cavity Fe2O3-rGO NRs under a physiological pH environment has been successfully realized, attributing to their high electron mobility and large specific surface area. Furthermore, the results of this work indicate that the glucose sensor prepared here has shown good repeatability and stability, which suggests its potential use in clinical detection.

Optical fiber sensor for curvature and temperature simultaneous measurement based on an anti-resonance mechanism

Based on an anti-resonance mechanism, we propose and experimentally demonstrate an optical fiber sensor for simultaneous measurement of curvature and temperature. The sensor structure consists of a hollow fiber and a twisted graded index multimode fiber spliced between two single-mode fibers. By using the twisted graded index multimode fiber to change the energy distribution of the mode field, not only improve the coupling efficiency of the sensor but also the sensitivity of the curvature measurement. The experimental results show that the intensity of the resonance peak based on the anti-resonance mechanism decreases with the increase of curvature, while the resonance peak wavelength is insensitive to curvature. In addition, due to the thermal expansion and thermo-optic effect of the fiber, the wavelength of the resonance peak is red-shifted with increasing temperature, and the intensity of the resonance peak is insensitive to temperature. Therefore, the intensity and wavelength signals of the resonance peak are monitored separately, and the simultaneous measurement of displacement and temperature is realized. The maximum sensitivity of the proposed curvature sensor can reach −2.84 dB/m−1, the dynamic range of curvature is 0 m−1 to 20.18 m−1, and the temperature sensitivity of 20 pm/∘C is realized. The proposed sensor has a compact structure, simple preparation, high coupling efficiency, and good repeatability, making it an ideal choice for simultaneous measurement of curvature and temperature in structural monitoring, mechanical manufacturing, and other fields.

Fully optically controlled Li-ion-mediated artificial vision reflection arc system

Artificial vision systems have been extensively reported for processing unstructured data, performing pattern recognition, and performing visual simulations. Photonic synapses have become the core of most artificial vision systems due to their advantages such as low energy consumption and ultra-fast signal transmission. However, their integration is difficult, the integration difficulty of photon synapses is higher than that of crossbar arrays. Moreover, due to the hybrid drive of light and external power supply, low energy consumption and weight update under full optical control cannot be achieved. Here, we designed a fully light-controlled multifunctional artificial vision reflex arc system. The solar cell and oscillator generate presynaptic voltage signals that are then transmitted to a lithium-ion-based memristor for the stimulation of different wavelengths and light intensities. The results show that our artificial vision system can distinguish wavelength and light intensity information and has good weight update symmetry and repeatability. Finally, the deformation experiment of the shape memory alloy demonstrated that the system can be used for controlling artificial muscles under different light signals.

Repeatability of Hydroacoustic Results versus Uncertainty in Assessing Changes in Ecological Status Based on Fish: A Case Study of Lake Widryńskie (Poland)

Assessments of changes in the ecological state of aquatic ecosystems are always burdened with uncertainty, which results from environmental reasons and poor repeatability of measurement results of elements enabling the assessment. This study determines the uncertainty related to the elements of the assessment of the hydroacoustic structure of fish communities’ (1) vertical target strength distribution (TS) in two-meter layers of water and (2) changes in the area where fish were recorded (which was determined on the basis of maps of their distribution in 2 m deep water layers). The object of this study was a lake (depth: 27 m) in which at the end of June 2016 the O2 concentration was &lt;1.4 mg L−1 below 8 m depth, which resulted in the accumulation of fish to a depth of 6 m. Hydroacoustic acquisition was carried out along transects arranged in the east–west (WE), north–south (NS), and zigzag (ZZ) directions in three repetitions. It was shown that the empirical probability of obtaining statistically different results was 2/9 when (1) Kendall’s τ coefficient, used to determine the similarity of the TS distribution, was less than 0.7—moderate correlation—and (2) fish occurrence areas in two cases (WE and ZZ on the third day of research) in layers 2–4 m and 4–6 m differed statistically significantly from the average area for all repetitions by 10–14% and 56–66% (p &lt; 0.05), respectively. The obtained results indicate quite good repeatability of acoustic measurements; however, in order to reduce the uncertainty, it is recommended that tests be conducted in this type of lake in three series of measurements.

The Development of a Sensitive Droplet Digital Polymerase Chain Reaction Test for Quantitative Detection of Goose Astrovirus

(1) Goose astrovirus (GAstV) is a novel emerging pathogen that causes significant economic losses in waterfowl farming. A convenient, sensitive, and specific detection method for GAstV in field samples is important in order to effectively control GAstV. Droplet digital polymerase chain reaction (ddPCR) is a novel, sensitive, good-precision, and absolute quantitation PCR technology which does not require calibration curves. (2) In this study, we developed a ddPCR system for the sensitive and accurate quantification of GAstV using the conserved region of the ORF2 gene. (3) The detection limit of ddPCR was 10 copies/µL, ~28 times greater sensitivity than quantitative real-time PCR (qPCR). The specificity of the test was determined by the failure of amplification of other avian viruses. Both ddPCR and qPCR tests showed good repeatability and linearity, and the established ddPCR method had high sensitivity and good specificity to GAstV. Clinical sample test results showed that the positive rate of ddPCR (88.89%) was higher than that of qPCR (58.33%). (4) As a result, our results suggest that the newly developed ddPCR method might offer improved analytical sensitivity and specificity in its GAstV measurements. The ddPCR could be widely applied in clinical tests for GAstV infections.

Flexible piezoelectric pressure sensor based on BaTiO3-CNT/RTV composite film with filter paper microstructures for monitoring hand motions

Flexible piezoelectric sensors are widely used in various applications such as physiological signal monitoring and human-computer interaction. The present study introduces a BaTiO3-CNT/RTV piezoelectric sensor fabricated using a filter paper template. It incorporates micro-scale fiber stacking and a 1% CNT doping in the microstructure, resulting in a notable enhancement of sensor sensitivity, increasing it from 0.07 V N−1 to 0.69 V N−1, representing an almost tenfold improvement. Furthermore, the study investigates the influence of affecting factors like the flexible substrate of the sensing film, thickness, and mass fractions of various materials on the output voltage. The sensor exhibits superior characteristics such as good repeatability under 5000 cyclic loads, high elongation at break, fast response (80 ms) and recovery times (90 ms), and good linearity. It also demonstrates outstanding sensitivity (12 mV/10°) when monitoring different finger bending states, enabling real-time, sensitive, and reliable hand motion tracking. This sensor holds promising prospects for future developments in the fields of intelligent grasping and sign language translation.

Quasi-distributed optical fiber hydrogen leakage detecting system based on bus chain topology structure

Micro-mirror optical fiber hydrogen sensors have the advantages of compact structure and fast demodulation speed. All-optical sensor networks consisting of micro-mirror optical fiber hydrogen sensors are essentially necessary across the hydrogen value chain. A bus chain topology structure hydrogen leakage detecting system based on micro-mirror sensors is proposed and experimentally demonstrated. A compensating optical path with constant power supply is introduced, and the power dissipation scheme is theoretically and experimentally proposed by designating the splitting ratios of the splitters array. By constructing such a network with twenty micro-mirror hydrogen sensors, the system has been experimentally verified with good repeatability and stability under different concentrations of hydrogen. By employing this bus chain topology strategy, a quasi-distributed optical fiber hydrogen leakage detection system with micro-mirror hydrogen sensors array is realized. It will provide a promising solution with high feasibility on multi-point leakage detecting in hydrogen infrastructures.

Evaluating fetal tricuspid and mitral annular plane systolic excursion (TAPSE and MAPSE) using spatiotemporal image correlation (STIC) M-mode.

This study aimed to evaluate the feasibility andreliability of tricuspid and mitral annular plane systolic excursion measurements (TAPSE and MAPSE) applying M-mode Spatiotemporal Image Correlation (STIC) technology in low-risk pregnancies. An initial retrospective pilot study was carried out to assess repeatability, followed by a larger mixed cross-sectional and prospective longitudinal study, both evaluating low-risk singleton pregnancies between 22+6 and 39+1 weeks. As only annuli capture was necessary, STIC acquisition parameters were set to the minimum volume angle of 15° and acquisition time of 7.5 s. A total of 330 volumes were analysed offline applying STIC M-mode. Acquisition rates were 96.9 % for TAPSE and 93.7 % MAPSE in the pilot study (n=32) and 98.0 % for both in the longitudinal study (n=102). Both study designsrevealed good repeatability for both sides of the heart, withhigher intraclass correlation coefficients (ICCs) for TAPSE (0.84-0.94) compared to MAPSE (0.80-0.88). Goodrepeatability was demonstrated for both sides of the heart, more so for TAPSE than MAPSE, with ICCs for TAPSE 0.84-0.91 and MAPSE 0.75-0.76, comparable to prior ICCs forSTIC repeatability. Modified STIC acquisition settings specifically tailored for capturing the longitudinal annular displacement may improve STIC TAPSE and MAPSE acquisition rates, optimising image quality for precise measurement and potentially bringing these modalities closer to clinical application.

High-sensitivity strain sensor based on an asymmetric tapered air microbubble Fabry-Pérot interferometer with an ultrathin wall

A Fabry-Pérot interferometer (FPI) with an asymmetric tapered structure and air microbubble with an ultrathin wall is designed for high-sensitivity strain measurement. The sensor contains an air microbubble formed by two single-mode fibers (SMF) prepared by fusion splicer arc discharge, and a taper is applied to one side of the air microbubble with a wall thickness of 3.6 µm. In this unique asymmetric structure, the microbubble is more easily deformed under stress, and the strain sensitivity of the sensor is up to 15.89 pm/µɛ as evidenced by experiments.The temperature sensitivity and cross-sensitivity of the sensor are 1.09 pm/°C and 0.069 µɛ/°C in the temperature range of 25-200°C, respectively, thus reducing the measurement error arising from temperature variations. The sensor has notable virtues such as high strain sensitivity, low-temperature sensitivity, low-temperature cross-sensitivity, simple and safe process preparation, and low cost. Experiments confirm that the sensor has good stability and repeatability, and it has high commercial potential, especially strain measurements in complex environments.

Magnetic solid-phase extraction of tetracyclines from milk using metal-organic framework MIL-101(Cr)-NH2 functionalised hydrophilic magnetic nanoparticles

Novel metal-organic framework MIL-101(Cr)-NH2 functionalised hydrophilic polydopamine-modified Fe3O4 magnetic nanoparticles (Fe3O4@PDA@MIL-101(Cr)-NH2) were synthesised and used as magnetic solid-phase extraction (MSPE) adsorbents for extracting tetracyclines (TCs) from milk samples. The integrated Fe3O4@PDA@MIL-101(Cr)-NH2 exhibited convenient magnetic separation and exceptional multi-target binding capabilities. Furthermore, the PDA coating significantly enhanced the hydrophilicity and extraction efficiency of the material, thereby facilitating the extraction of trace TCs. Various factors affecting MSPE, such as adsorbent dosage, extraction time, pH value, and desorption conditions, were optimised. The developed MSPE method coupled with high-performance liquid chromatography demonstrated good linearity (R2 ≥ 0.9989), acceptable accuracy (82.2%–106.1%), good repeatability (intra-day precision of 0.8%–4.7% and inter-day precision of 1.1%–4.5%), low limits of detection (2.18–6.25 μg L−1), and low limits of quantification (6.54–18.75 μg L−1) in TCs detection. The approach was successfully used for the quantification of trace TCs in real milk samples.

Establishment of a real-time fluorescent quantitative PCR detection method and phylogenetic analysis of BoAHV-1

BackgroundInfectious bovine rhinotracheitis (IBR), caused by Bovine alphaherpesvirus-1 (BoAHV-1), is an acute, highly contagious disease primarily characterized by respiratory tract lesions in infected cattle. Due to its severe pathological damage and extensive transmission, it results in significant economic losses in the cattle industry. Accurate detection of BoAHV-1 is of paramount importance. In this study, we developed a real-time fluorescent quantitative PCR detection method for detecting BoAHV-1 infections. Utilizing this method, we tested clinical samples and successfully identified and isolated a strain of BoAHV-1.1 from positive samples. Subsequently, we conducted a genetic evolution analysis on the isolate strain’s gC, TK, gG, gD, and gE genes.ResultsThe study developed a real-time quantitative PCR detection method using SYBR Green II, achieving a detection limit of 7.8 × 101 DNA copies/μL. Specificity and repeatability analyses demonstrated no cross-reactivity with other related pathogens, highlighting excellent repeatability. Using this method, 15 out of 86 clinical nasal swab samples from cattle were found to be positive (17.44%), which was higher than the results obtained from conventional PCR detection (13.95%, 12/86). The homology analysis and phylogenetic tree analysis of the gC, TK, gG, gD, and gE genes of the isolated strain indicate that the JL5 strain shares high homology with the BoAHV-1.1 reference strains. Amino acid sequence analysis revealed that gC, gE, and gG each had two amino acid mutations, while the TK gene had one synonymous mutation and one H to Y mutation, with no amino acid mutations observed in the gD gene. Phylogenetic tree analysis indicated that the JL5 strain belongs to the BoAHV-1.1 genotype and is closely related to American strains such as C33, C14, and C28.ConclusionsThe established real-time fluorescent quantitative PCR detection method exhibits good repeatability, specificity, and sensitivity. Furthermore, genetic evolution analysis of the isolated BoAHV-1 JL-5 strain indicates that it belongs to the BoAHV-1.1 subtype. These findings provide a foundation and data for the detection, prevention, and control Infectious Bovine Rhinotracheitis.