Blocking Buffer Research Articles

Mechanism of Status Evaluation for Water Blocking Buffer Layer of High-Voltage Cable Based on Characteristic Gas Analysis

Mechanism of Status Evaluation for Water Blocking Buffer Layer of High-Voltage Cable Based on Characteristic Gas Analysis

Development and validation of enzyme-linked immunosorbent assay for anti-mouse pertussis immunoglobulin G using international reference anti-Bordetella pertussis mouse serum NIBSC 97/642.

In this study, an in-house enzyme-linked immunosorbent assay (ELISA) was developed and validated. The titer of ELISA was calculated using the reference line (RFL) method based on the standard curve drawn using the international reference anti-mouse serum NIBSC (National Institute for Biological Standards and Control) 97/642. In the development step, signal to noise was depicted to select the buffers that showed the most appropriate ratio. In the validation step, standard range, precision, dilution linearity, and specificity were confirmed, and RFL and parallel line (PLL) methods were compared in precision and dilution linearity. Coating concentration for plate was achieved at 0.1 µg/mL for pertussis toxin (PT), 0.15 µg/mL for filamentous hemagglutinin antigen (FHA), and 0.25 µg/mL for pertactin (PRN). The signal to noise ratio was 22.02 for PT, 14.93 for FHA, and 8.02 for PRN with 0.25% goat serum in phosphate-buffered saline (PBS) as a dilution buffer, and 2% skim milk in PBS as a blocking buffer. Based on the precision results, we assessed the lower limit of quantification by 1, 0.2, and 1.5 EU/mL concentration for PT, FHA, and PRN which met the ICH (International Council for Harmonization) M10 criteria of a 25% accuracy and total error of 40%. In specificity, homologous serum was spiked into heterologous serum and the accuracy met the criteria. There was no difference in the results between RFL and PLL calculations (p-value=0.3207 for PT, 0.7394 for FHA, 0.2109 for PRN). ELISA validated with RFL calculation method in this study is a relatively accurate assay for mouse humoral immunogenicity test.

Development and Validation of an Enzyme-Linked Immunosorbent Assay-Based Protocol for Evaluation of Respiratory Syncytial Virus Vaccines.

Recently, respiratory syncytial virus (RSV) vaccines based on the prefusion F (pre-F) antigen were approved in the United States. We aimed to develop an enzyme-linked immunosorbent assay (ELISA)-based protocol for the practical and large-scale evaluation of RSV vaccines. Two modified pre-F proteins (DS-Cav1 and SC-TM) were produced by genetic recombination and replication using an adenoviral vector. The protocol was established by optimizing the concentrations of the coating antigen (pre-F proteins), secondary antibodies, and blocking buffer. To validate the protocol, we examined its accuracy, precision, and specificity using serum samples from 150 participants across various age groups and the standard serum provided by the National Institute of Health. In the linear correlation analysis, coating concentrations of 5 and 2.5 μg/mL of DS-Cav1 and SC-TM showed high coefficients of determination (r > 0.90), respectively. Concentrations of secondary antibodies (alkaline phosphatase-conjugated anti-human immunoglobulin G, diluted 1:2000) and blocking reagents (5% skim milk/PBS-T) were optimized to minimize non-specific reactions. High accuracy was observed for DS-Cav1 (r = 0.90) and SC-TM (r = 0.86). Further, both antigens showed high precision (coefficient of variation < 15%). Inhibition ELISA revealed cross-reactivity of antibodies against DS-Cav1 and SC-TM, but not with the attachment (G) protein.

Design and fabrication of a competitive lateral flow assay using gold nanoparticle as capture probe for the rapid and on-site detection of penicillin antibiotic in food samples

Lateral flow assays (LFAs) are among the utmost cost-efficient, paper-based point-of-care (POC) diagnostic devices. Herein, we have reported the fabrication of a competitive LFA for on-site detection of penicillin. Various parameters such as Ab concentration for conjugation, Pen-BSA conjugate concentration, pore size of membrane, and blocking buffer were standardised for the fabrication of LFA. Different concentrations of penicillin (1 pM-1 mM) were added to the sample pad to observe the color intensity. The visual detection limit (LOD) achieved from the LFA was 10 nM for Penicillin that correlated with the LOD calculated from the ‘ColorGrab’ colorimeter application. Additionally, LFA showed insignificant cross reactivity with other β-lactam antibiotics and were also validated with spiked food samples such as milk, meat and egg. Hence, the fabricated LFA can be successfully utilised for the POC detection of penicillin in food samples on large scale.

An optimization of blocking agents for designing reliable electrochemical biosensors for ovarian cancer

Here, we report development of an electrochemical biosensor for ovarian cancer diagnosis using an optimized blocking agent for reliable performance of the device. The sensitivity, specificity and reproducibility of the device are negatively impacted by non-specific binding of molecules present in sample - proteins, DNA, RNA, small drug molecules, hormones etc, other than the analyte of interest bind to biorecognition molecule on the biosensor surface. Electrochemical biosensor was fabricated by functionalizing carbon screen printed electrodes with citrate-reduced gold nanoparticles followed by immobilization of 5′-amine modified ssDNA probe against miRNA 204 (ovarian can biomarker). To avoid or minimize non-specific binding, blocking reagent was optimized. Blocking reagents and methods are typically chosen in an empirical manner since a single standardized procedure has not been determined suitable for all applications. This paper presents the commonly used blocking buffers with varying molecular weights to minimize the non-specific binding process on the surface of the electrode. Here, three different blocking buffers are used - Bovine serum albumin (BSA; MW: 66430.3 kDa), Gelatin (MW: 40 kDa), and Polyethylene Glycol (PEG MW 4 kDa and 6 kDa). Additionally, different surfactants like tween20, Tritonx100, and HEPES buffer are used for preparation of blocking agents. Further, to evaluate the performance of different blocking agents, the sample of miRNA-204 is spiked in 0.01 M Phosphate buffer saline (PBS) and fetal bovine serum (FBS) were tested using chroamperometry. The difference in the saturation current was recorded for both curves obtained for 0.01 M PBS and FBS. Optimum blocking buffer 1% Gelatin in Tween20 was found to be giving negligible nonspecific binding for DNA based ovarian cancer biosensor.

BD Pharmingen™ Leukocyte Blocking Buffer reduces nonspecific binding of cyanine-like dye conjugates to monocytes and other leukocytes

Abstract Nonspecific binding of monocytes to certain cyanine-like dye conjugated antibodies that are commonly used in flow cytometry for cell surface staining is well known. Although lesser in extent than monocytes, other leukocyte subsets can also exhibit nonspecific binding to cyanine-like dye conjugates. This poses a challenge for multicolor flow cytometric analysis utilizing these cyanine-like dye conjugates as dye-mediated nonspecific binding can mask the real expression. Here we demonstrate that the BD Pharmingen™ Leukocyte Blocking Buffer, a blocking buffer solution, eliminates nonspecific binding of cyanine-like dye conjugates to monocytes and other leukocyte populations. In this study we show that human whole blood stained with PE-Cy5, PE-Cy7, APC-Cy7, APC-H7, PE-CF594 and other tandem-dye conjugated antibodies in the presence of this new buffer exhibits the appropriate staining profile of these antibodies to monocytes and other leukocyte subsets. We also observed that nonspecific binding of cyanine-like dye conjugated antibodies to myeloid and lymphoid subsets in mouse lymphoid tissues was diminished. BD Pharmingen™ Leukocyte Blocking Buffer, while reducing nonspecific cyanine-like dye conjugates binding, did not impact the desirable antibody-mediated specific staining. Our data also show that cell viability is not impacted. For Research Use Only. Not for use in diagnostic or therapeutic procedures. BD and Pharmingen are trademarks of Becton, Dickinson and Company or its affiliates. © 2023 BD. All rights reserved. NPM-0263 (v1.0) 1222 CF is a trademark of Biotium, Inc. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.

A Stream-Aware MPQUIC Scheduler for HTTP Traffic in Mobile Networks

A QUIC (Quick UDP Internet Connections) protocol is designed to improve Hypertext Transfer Protocol (HTTP) traffic and carries a non-negligible portion of the traffic in the current Internet. As its extension, Multipath QUIC (MPQUIC) provides higher bandwidth and smoother network handover by using multiple network interfaces simultaneously. However, to improve HTTP traffic, there are still some issues not yet carefully addressed in the existing MPQUIC, and packet scheduling is a vital one among the issues. Specifically, existing methods fail to respond to the stream prioritization of HTTP Version 2 (HTTP/2), leading to unsatisfying web page load performance. Besides, managing asymmetric and dynamic network paths is also a challenging issue, which may result in Head-of-Line (HoL) blocking and excessive buffer usage if not effectively handled. In this paper, we present a stream-aware per-packet scheduler, HoL Blocking Eliminating Scheduler (HBES), to improve the performance of MPQUIC in mobile networks. Firstly, HBES provides a fair allocation of aggregated bandwidth for different streams based on their priority. Then, it keeps stream data arriving at the receiver in order by estimating packet arrival time to mitigate HoL blocking and excessive buffer usage. We implement HBES and evaluate its performance in various network scenarios. Experimental results verify the superiority of HBES in reducing stream completion time and buffer occupation over those existing MPQUIC schedulers.

Influence of bovine and human serum albumin on the binding kinetics of biomolecular interactions.

Bovine serum albumin (BSA) containing buffers are the standard blocking buffer in biosensing, yet human serum is the intended application for most clinical sensors. However, the effect of human serum albumin (HSA) on binding assays remains underexplored. A simple and well-studied assay (human IgG/goat anti-human IgG) was investigated with a surface plasmon resonance (SPR) sensor to address this fundamental question in sensing. Calibrations were performed with buffers containing various concentrations of bovine or human serum albumin, as well as full and diluted bovine or IgG-depleted human serum. It was found that HSA or human serum, but not BSA or bovine serum, significantly affected the SPR shift and binding constants of the assay. Interestingly, large differences were also observed depending on whether the animal or human antibody was immobilized on the SPR chip for detection, highlighting that matrix protein/analyte/receptor interactions play a significant role in the response. We find that the interaction of soluble HSA with human IgG interferes with the recognition region, affecting the binding constant, and thus results obtained in BSA are not necessarily applicable to clinical samples or in vivo conditions. We also clearly demonstrate why a minimum dilution of 1 : 10 is often required in SPR assays to remove most background effects. Taken together, these results show that: (1) BSA does not affect the binding constant between antibodies and thus serves its purpose well when only surface blocking is intended, (2) HSA is an adequate surrogate for human serum in assay optimization, and (3) blocking buffers should be prepared with HSA in the optimization steps of assays to be translated to human blood or serum.

Study on Technical Parameters of X-ray Stereoscopic Imaging of Ablative Defects for Water Blocking Buffer Layer in High Voltage Cable

In this paper, the technical parameters of the X-ray stereoscopic imaging of the ablative defects of water blocking buffer layer in the high-voltage cable are studied by theoretical analysis and laboratory experiment. The works give technical recommendations about the voltage, current, and sampling volume, which provide a reference for the standardization of the technical parameters of X-ray stereoscopic imaging of ablative defects of the water-blocking buffer layer in the high-voltage cable.

Abstract 15484: Distinct Subcellular Localizations of Non-Muscle Myosin IIB and IIC in Cardiomyocytes

Introduction: Non-muscle myosin (NMII) contraction plays a role in generating cellular tension and its role in mediating myocardial hypertrophy is emerging. Non-muscle myosin is present in cardiomyocytes in two isoforms NMIIB and NMIIC. Prior literature suggests that NMIIB and NMIIC are found in the intercalated disc, while NMIIB is found in the Z discs. Recently, PKC was implicated in medating stiffness-dependent cardiomyocyte hypertrophy through NMII. Moreover, PKC and NMII were found to be upregulated at the costameres in heart disease. The respective role of NMIIB and NMIIC in mediating cardiac hypertrophy remains unknown. Methods: Cardiac tissues from 10w male C57BL6/J wildtype and Myh14-GFP mice were fixed in BE40 solution (70% ethanol, 1% glycerol, 0.5% glcial acetic acid, and 28.5% 0.5 x PBS) for 3 days. Tissues were washed in 70% ethanol and made into paraffin embedded sections. Sections were heated for 10 min at 50C, deparaffinized, washed in ethanol and rinsed in PBS. Antigen retrieval was performed by heating for 10 min in TET buffer (Tris-EDTA, pH9.0 and 0.05% Tween 20). After PBS rinse, sections were incubated in blocking buffer (3% NGS, 1% BSA, 0.15% Triton X 100, PBS) for 1 hour at RT. Then sections are incubated in primary antibodies in the blocking buffer for 1 hour at RT. After PBS rinse, sections were incubated in secondary antibodies in the blocking buffer for 1 hour at RT. After PBS rinse, sections were mounted in Vectashield with DAPI. Images were captured using a Nikon A1 confocal microscopy with 60X oil lens and 2X zoom. Results: NMIIB is expressed in a subset of intercalated discs and Z lines. NMIIC is expressed in a subset of intercalated discs and costameres. Phospho-Myosin light chain (P-MLC) partially co-localizes with NMIIC in the intercalated discs but is not found in costameres. Conclusions: NMIIB and NMIIC display distinct subcellular localization, including within the intercalated discs. Only NMIIB is found at Z lines. Only NMIIC is found at costameres. P-MCL is only found in the intercalated discs. These findings suggest differential regulation and function of NMIIB and NMIIC. Further characterization of specific subcellular NMIIB and NMIIC function may elucidate their respective roles in tensional homeostasis in the cardiomyocyte.

A Flow Cytometry-based Cell Surface Protein Binding Assay for Assessing Selectivity and Specificity of an Anticancer Aptamer.

A key challenge in developing an anticancer aptamer is to efficiently determine the selectivity and specificity of the developed aptamer to the target protein. Due to its several advantages over monoclonal antibodies, aptamer development has gained enormous popularity among cancer researchers. Systematic evolution of ligands by exponential enrichment (SELEX) is the most common method of developing aptamers specific for proteins of interest. Following SELEX, a quick and efficient binding assay accelerates the process of identification, confirming the selectivity and specificity of the aptamer. This paper explains a step-by-step flow cytometric-based binding assay of an aptamer specific for epithelial cellular adhesion molecule (EpCAM). The transmembrane glycoprotein EpCAM is overexpressed in most carcinomas and plays roles in cancer initiation, progression, and metastasis. Therefore, it is a valuable candidate for targeted drug delivery to tumors. To evaluate the selectivity and specificity of the aptamer to the membrane-bound EpCAM, EpCAM-positive and -negative cells are required. Additionally, a non-binding EpCAM aptamer with a similar length and 2-dimensional (2D) structure to the EpCAM-binding aptamer is required. The binding assay includes different buffers (blocking buffer, wash buffer, incubation buffer, and FACS buffer) and incubation steps. The aptamer is incubated with the cell lines. Following the incubation and washing steps, the cells will be evaluated using a sensitive flow cytometry assay. Analysis of the results shows the binding of the EpCAM-specific aptamer to EpCAM-positive cells and not the EpCAM-negative cells. In EpCAM-positive cells, this is depicted as a band shift in the binding of the EpCAM aptamer to the right compared to the non-binding aptamer control. In EpCAM-negative cells, the corresponding bands of EpCAM-binding and -non-binding aptamers overlap. This demonstrates the selectivity and specificity of the EpCAM aptamer. While this protocol is focused on the EpCAM aptamer, the protocol is applicable to other published aptamers.

Integrated quay crane–shuttle vehicle scheduling problem capacitated by apron buffer

This paper investigates an integrated quay crane (QC)–shuttle vehicle (SV) scheduling problem (SP) with capacitated apron buffer, which is subject to practical constraints on non-zero travelling time, precedence relationships between container-handling jobs, QC non-interference, safety margin, blocking and apron buffer capacity. The objective is to minimise the makespan of vessel handling by simultaneous optimisation of bay-to-QC assignment, the sequences of bays handled by QCs, the sequences of jobs handled by QCs, job-to-SV assignment and the sequences of jobs handled by SVs. To achieve this, a mixed-integer linear programming model is first formulated to solve the SP. The relationships of decision variables and model constraints are then analysed to generate two sub-models: assignment based and sequence based. The former sub-model is developed from the proposed model by pre-determining the assignment of bay-to-QC and job-to-SV, and the latter is developed from the proposed model by pre-determining the sequences of jobs handled by each QC and each SV. Hybrid genetic algorithms with the two sub-models are proposed for numerical experiments. Practical implications of this investigation for apron buffer zone design are discussed.

An FPGA‐based JPEG preprocessing accelerator for image classification

The FPGA-based image classification accelerator has achieved success in many practical applications. However, most accelerators focus on solving the problem of convolution computation efficiency. End-to-end image classification involves many non-convolutional operations, which can also become performance bottlenecks. Therefore, the authors propose an FPGA-based JPEG preprocessing accelerator, which can accelerate non-convolution operations of JPEG before feature extraction. To improve throughput and energy efficiency, four hardware structures are adopted in the design: 1) adaptive image block; 2) fast IDCT; 3) image block buffer; and 4) image block self-location. The proposed design is evaluated on Xilinx XCZU7EV. The authors compare it with the optimized implementation of CPU and GPU, and the energy efficiency is improved by 23.07 times and 4.21 times, respectively. The throughput is 2.52 times better than the CPU implementation. And the authors demonstrate its practicality through a case study of image classification. These experimental results demonstrate its superior performance in terms of throughput and energy efficiency.

Induction of broadly reactive influenza antibodies increases susceptibility to autoimmunity.

Infection and vaccination repeatedly expose individuals to antigens that are conserved between influenza virus subtypes. Nevertheless, antibodies recognizing variable influenza epitopes greatly outnumber antibodies reactive against conserved epitopes. Elucidating factors contributing to the paucity of broadly reactive influenza antibodies remains a major obstacle for developing a universal influenza vaccine. Here, we report that inducing broadly reactive influenza antibodies increases autoreactive antibodies in humans and mice and exacerbates disease in four distinct models of autoimmune disease. Importantly, transferring broadly reactive influenza antibodies augments disease in the presence of inflammation or autoimmune susceptibility. Further, broadly reactive influenza antibodies spontaneously arise in mice with defects in B cell tolerance. Together, these data suggest that self-tolerance mechanisms limit the prevalence of broadly reactive influenza antibodies, which can exacerbate disease in the context of additional risk factors.

An Energy-Efficient DRAM Cache Architecture for Mobile Platforms With PCM-Based Main Memory

A long battery life is a first-class design objective for mobile devices, and main memory accounts for a major portion of total energy consumption. Moreover, the energy consumption from memory is expected to increase further with ever-growing demands for bandwidth and capacity. A hybrid memory system with both DRAM and PCM can be an attractive solution to provide additional capacity and reduce standby energy. Although providing much greater density than DRAM, PCM has longer access latency and limited write endurance to make it challenging to architect it for main memory. To address this challenge, this article introduces CAMP, a novel DRAM c ache a rchitecture for m obile platforms with P CM-based main memory. A DRAM cache in this environment is required to filter most of the writes to PCM to increase its lifetime, and deliver highest efficiency even for a relatively small-sized DRAM cache that mobile platforms can afford. To address this CAMP divides DRAM space into two regions: a page cache for exploiting spatial locality in a bandwidth-efficient manner and a dirty block buffer for maximally filtering writes. CAMP improves the performance and energy-delay-product by 29.2% and 45.2%, respectively, over the baseline PCM-oblivious DRAM cache, while increasing PCM lifetime by 2.7×. And CAMP also improves the performance and energy-delay-product by 29.3% and 41.5%, respectively, over the state-of-the-art design with dirty block buffer, while increasing PCM lifetime by 2.5×.

Development of an indirect ELISA method based on the VP4 protein for detection antibody against duck hepatitis A virus type 1

In Picornavirus, the VP4 gene is located inside the viral capsid, but the antibodies it produces have neutralizing activity. At the same time, we know that the VP4 gene is relatively conserved among the four structural proteins, which makes the usage VP4 protein-based antigen potentially meaningful. Therefore, we used purified duck hepatitis A virus type 1 (DHVA-1) recombinant VP4 protein as the coating antigen to establish an indirect method. The optimal antigen, serum and enzyme-labeled antibody dilutions were 1:400 (3.375 μg/mL), 1:80 and 1:1600, respectively. The optimal blocking buffer was 1% skimmed milk powder. The cutoff value was determined to be 0.203, and the analytical sensitivity was 1:1600. The established ELISA method has good specificity, repeatability and sensitivity. It has a high coincidence rate of 70.8 % with the DHVA-1 as the coating antigen. So, it can be used for the detection of DHAV-1 serum antibodies.

Dynamic response of reinforced concrete sheds against the impact of rock block with different shapes and angles

This study aimed to quantitatively identify the influence of the block impact angle and block shape on the impact effect of reinforced concrete (RC) sheds. The smooth particle hydrodynamic (SPH) method and finite element method (FEM) were coupled and used to solve the simulation difficulty of large deformation of the sand buffer layer in the RC shed. SPH was used to simulate the sand buffer layer in the impact area that experienced large deformation. Concrete, reinforced, bedrock, block and sand buffer layer in the non-impact area were simulated by FEM. The accuracy of the coupled model was verified by the full-scale test data. Finally, the impact forces and the dynamic responses of the RC shed were analyzed, focusing on the effects of the block impact angle and shape. The numerical results show that the coupled SPH-FEM method is effective for simulating how block impacts the RC shed. The block impact angle and shape can significantly influence the normal and tangential impact forces. The design of the RC shed based on the assumption of blocks as free-falling spherical projectiles can lead to inaccuracy in some impact scenarios, and an appropriate correction was put forward in available empirical calculating formulae to include the effect of the irregular shape and impact angle of the block. The sand buffer layer dissipates a large part of the energy, which accounts for at least 58% of the initial impact energy.

Neutralization antibody titers against SARS-CoV-2 in an infant born to a mother with COVID-19

Neutralization antibody titers against SARS-CoV-2 in an infant born to a mother with COVID-19

A Comparative Analysis of Delay Propagation on Departure and Arrival Flights for a Chinese Case Study

In recent years, flight delay costs the air transportation industry millions of dollars and has become a systematic problem. Understanding the behavior of flight delay is thus critical. This paper focuses on how flight delay is affected by operation-, time-, and weather-related factors. Different econometric models are developed to analyze departure and arrival delay. The results show that compared to departure delay, arrival delay is more likely to be affected by previous delays and the buffer effect. Block buffer presents a reduction effect seven times greater than turnaround buffer in terms of flight delays. Departure flights suffer more delays from convective weather than arrival flights. Convective weather at the destination airport for flight delay has a greater impact than at the original airport. In addition, sensitivity analysis of flight delays from an aircraft utilization perspective is conducted. We find that the effect of delay propagation on flight delay differs by aircraft utilization. This impact on departure delay is greater than the impact on arrival delay. In general, specific to the order of flights, the previous delay increases the impact on flight on-time performance as a flight flies a later leg. Buffer time has opposite effects on departure and arrival delay, with the order increasing. A decrease in buffer time with the order increasing, however, still has a greater reduction effect on departure delay than arrival delay. Specific to the number of flights operated by an aircraft, the more flights an aircraft flies in a day, the more the on-time performance of those flights will suffer from the previous delay and buffer time generally.

Establishment and evaluation of an indirect ELISA for detection of antibodies to goat Klebsiella pneumonia

BackgroundKlebsiella pneumonia, a Gram-negative bacterium belonging to the genus Enterobacter, causes many human and livestock diseases. Notably, infected goats may develop pneumonia, septicemia, which can lead to occasional death, resulting in great economic losses in goat-farming industry. However, there are little systematic methods for detection of goat Klebsiella pneumoniae in livestock production.ResultsIn this study, we developed a Klebsiella pneumoniae goat polyclonal antibody and established an indirect ELISA method to detect the Klebsiella pneumoniae. After screening and optimizing the conditions for detection, we determined the optimal working dilutions of the coated-bacterial antigen, the polyclonal antibody, and the enzyme-labeled secondary antibody that were 1:800 (2.99 × 107 CFU/ml), 1:6400, and 1:5000, respectively. The optimal condition of coating and blocking were both 4 °C for 12 h. The optimal dilution buffers of bacterial antigen, the antibodies, and the blocking buffer were 0.05 mol/L carbonate buffer, 1% BSA phosphate buffer, and 1.5% BSA carbonate buffer, respectively. The cut-off value was determined to be 0.28, and the analytical sensitivity was 1:800 (dilution of a positive sample). Furthermore, there was no cross-reaction between the coated antigen and goat serum positive for antibodies against other bacteria, indicating that indirect ELISA could detect Klebsiella pneumoniae specifically in most cases. The average coefficients of variation of intra-assay and inter-assay were 4.37 and 5.17% indicating favorable reproducibility of indirect ELISA. In the detection of clinical veterinary samples, the positive rate of indirect ELISA was 6.74%, higher than that of conventional agglutination assays.ConclusionsTaken together, we successfully established an indirect ELISA method for detecting antibodies against Klebsiella pneumoniae in goats, which can be applied in production.