scholarly journals Use of oral fluids to detect anti Lawsonia intracellularis antibodies in experimentally infected pigs

2020 ◽  
Vol 40 (12) ◽  
pp. 970-976
Author(s):  
Michelle P. Gabardo ◽  
José Paulo H. Sato ◽  
Talita P. Resende ◽  
Luisa V.A. Otoni ◽  
Lucas A. Rezende ◽  
...  
Keyword(s):  
Oral Fluid ◽  
Immunoglobulin A ◽  
Standard Technique ◽  
High Specificity ◽  
Post Inoculation ◽  
Lawsonia Intracellularis ◽  
Serum Samples ◽  
Diagnostic Assays ◽  
Specificity And Sensitivity ◽  
Moderate Sensitivity

ABSTRACT: Several pathogens and antibodies derived from serum or produced in tissues associated with the oral cavity are present in the oral fluid (OF). Considering the applicability of this alternative sample, recent studies in veterinary medicine have tested OF as a replacement for serum in diagnostic assays. The aim of this study was to standardize the immunoperoxidase monolayer assay (IPMA) to detect anti-Lawsonia intracellularis immunoglobulin A (IgA) and immunoglobulin G (IgG) in OF samples from experimentally infected pigs. Sixty-two pigs were divided into two groups: control (T1, n=30) and inoculated with L. intracellularis (T2, n=32). Blood, OF and fecal samples were collected at 0, 7, 14, 21, 28 and 42 days post-inoculation (dpi). Some adaptations of the standard technique for serum were made to IPMA for the detection of IgA and IgG in OF. The IPMA showed high specificity and sensitivity for serum samples and high specificity and moderate sensitivity for the detection of IgA and IgG in OF. There was high agreement between the results of serum IgG and OF IgA and IgG. Based on our results, oral fluid samples may be used for the evaluation and determination of anti-L. intracellularis antibodies in pigs, but not for individual diagnosis of swine proliferative enteropathy.

scholarly journals Serum MicroRNA Signature as a Diagnostic and Therapeutic Marker in Patients with Psoriatic Arthritis

2020 ◽  
Vol 47 (12) ◽  
pp. 1760-1767
Author(s):  
Sarah M. Wade ◽  
Trudy McGarry ◽  
Siobhan C. Wade ◽  
Ursula Fearon ◽  
Douglas J. Veale
Keyword(s):  
Psoriatic Arthritis ◽  
Characteristic Curve ◽  
High Specificity ◽  
Serum Samples ◽  
Rna Molecules ◽  
Serum Mirna ◽  
Serum Microrna ◽  
Specificity And Sensitivity ◽  
European League Against Rheumatism ◽  
Noninvasive Markers

ObjectiveMicroRNA (miRNA) are small endogenous regulatory RNA molecules that have emerged as potential therapeutic targets and biomarkers in autoimmunity. Here, we investigated serum miRNA levels in patients with psoriatic arthritis (PsA) and further assessed a serum miRNA signature in therapeutic responder versus nonresponder PsA patients.MethodsSerum samples were collected from healthy controls (HC; n = 20) and PsA patients (n = 31), and clinical demographics were obtained. To examine circulatory miRNA in serum from HC and PsA patients, a focused immunology miRNA panel was analyzed utilizing a miRNA Fireplex assay (FirePlex Bioworks Inc.). MiRNA expression was further assessed in responders versus nonresponders according to the European League Against Rheumatism response criteria.ResultsSix miRNA (miR-221-3p, miR-130a-3p, miR-146a-5p, miR-151-5p, miR-26a-5p, and miR-21-5p) were significantly higher in PsA compared to HC (all P < 0.05), with high specificity and sensitivity determined by receiver-operating characteristic curve analysis. Analysis of responder versus nonresponders demonstrated higher baseline levels of miR-221-3p, miR-130a-3p, miR-146a-5p, miR-151-5p, and miR-26a-5p were associated with therapeutic response.ConclusionThis study identified a 6-serum microRNA signature that could be attractive candidates as noninvasive markers for PsA and may help to elucidate the disease pathogenesis.

scholarly journals Detection of Actinobacillus pleuropneumoniae ApxIV toxin antibody in serum and oral fluid specimens from pigs inoculated under experimental conditions

2017 ◽  
Vol 61 (2) ◽  
pp. 163-171 ◽  
Author(s):  
Wendy González ◽  
Luis G. Giménez-Lirola ◽  
Ashley Holmes ◽  
Sergio Lizano ◽  
Christa Goodell ◽  
...  
Keyword(s):  
Oral Fluid ◽  
Serum Antibody ◽  
Population Monitoring ◽  
Actinobacillus Pleuropneumoniae ◽  
Antibody Responses ◽  
Antibody Detection ◽  
Post Inoculation ◽  
Serum Samples ◽  
Experimental Conditions ◽  
And Control

AbstractIntroduction:The prevention and control ofActinobacillus pleuropneumoniaein commercial production settings is based on serological monitoring. Enzyme-linked immunosorbent assays (ELISAs) have been developed to detect specific antibodies against a variety ofA. pleuropneumoniaeantigens, including long-chain lipopolysaccharides (LPS) and the ApxIV toxin, a repeats-in-toxin (RTX) exotoxin unique toA. pleuropneumoniaeand produced by all serovars. The objective of this study was to describe ApxIV antibody responses in serum and oral fluid of pigs.Material and Methods:Four groups of pigs (six pigs per group) were inoculated withA. pleuropneumoniaeserovars 1, 5, 7, or 12. Weekly serum samples and daily oral fluid samples were collected from individual pigs for 56 days post inoculation (DPI) and tested by LPS and ApxIV ELISAs. The ApxIV ELISA was run in three formats to detect immunlgobulins M, G, and A (IgM, IgG and IgA) while the LPS ELISA detected only IgG.Results:All pigs inoculated withA. pleuropneumoniaeserovars 1 and 7 were LPS ELISA serum antibody positive from DPI 14 to 56. A transient and weak LPS ELISA antibody response was observed in pigs inoculated with serovar 5 and a single antibody positive pig was observed in serovar 12 at ≥35 DPI. Notably, ApxIV serum and oral fluid antibody responses in pig inoculated with serovars 1 and 7 reflected the patterns observed for LPS antibody, albeit with a 14 to 21 day delay.Conclusion:This work suggests that ELISAs based on ApxIV antibody detection in oral fluid samples could be effective in population monitoring forA. pleuropneumoniae.

scholarly journals INDIRECT ELISA FOR DETECTION OF ANTIBODIES TO FMDV NON-STRUCTURAL PROTEINS IN PORCINE SERA

2018 ◽  
pp. 13-20
Author(s):  
A. S. Yakovleva ◽  
A. V. Kanshina ◽  
A. V. Scherbakov
Keyword(s):  
Indirect Elisa ◽  
High Sensitivity ◽  
High Specificity ◽  
Structural Proteins ◽  
Post Inoculation ◽  
Nonstructural Proteins ◽  
Diagnostic Specificity ◽  
Fmd Virus ◽  
Specificity And Sensitivity ◽  
Sensitivity Specificity

An indirect variant of ELISA used for detection of antibodies to nonstructural proteins of the FMD virus in porcine blood sera was developed. The results of the validation showed that the developed method is characterized by high sensitivity, specificity and reproducibility. When testing the blood serum panel obtained from experimentally infected animals, the method allowed to detect antibodies to FMD virus in 7 of 18 sera collected on day 6 post inoculation, in 13 of 19 sera – on day 7 post inoculation, in 16 of 19 sera – on day 8 post inoculation and in all 76 sera obtained on days 9–12 post inoculation. The diagnostic specificity of 3AB-ELISA was 100% when testing 100 knowingly negative blood sera from pigs imported to Russia from Norway. High specificity and sensitivity of the method, established during the development of the method, are confirmed in the course of routine diagnostic tests.

scholarly journals Evaluation of SARS-CoV-2 Neutralizing Antibodies Using a Vesicular Stomatitis Virus Possessing SARS-CoV-2 Spike Protein

2020 ◽  
Author(s):  
Hideki Tani ◽  
Long Tan ◽  
Miyuki Kimura ◽  
Yoshihiro Yoshida ◽  
Hiroshi Yamada ◽  
...  
Keyword(s):  
Virus Infection ◽  
Whole Blood ◽  
Neutralizing Antibodies ◽  
High Specificity ◽  
Immunofluorescence Assay ◽  
Hospitalized Patients ◽  
Pseudotyped Virus ◽  
Serum Samples ◽  
Live Virus ◽  
Specificity And Sensitivity

Abstract Background:SARS-CoV-2 is a novel coronavirus that emerged in 2019 and is now classified in the genus Coronavirus with closely related SARS-CoV. SARS-CoV-2 is highly pathogenic in humans and is classified as a biosafety level (BSL)-3 pathogen, which makes manipulating it relatively difficult due to its infectious nature. Methods:To circumvent the need for BSL-3 laboratories, an alternative assay was developed that avoids live virus and instead uses a recombinant VSV expressing luciferase and possesses the full length or truncated spike proteins of SARS-CoV-2. Furthermore, to measure SARS-CoV-2 neutralizing antibodies under BSL2 conditions, a chemiluminescence reduction neutralization test (CRNT) for SARS-CoV-2 was developed. The neutralization values of the serum samples collected from hospitalized patients with COVID-19 or SARS-CoV-2 PCR-negative donors against the pseudotyped virus infection evaluated by the CRNT were compared with antibody titers determined from an immunofluorescence assay (IFA). Results:The CRNT, which used whole blood collected from hospitalized patients with COVID-19, was also examined. As a result, the inhibition of pseudotyped virus infection was specifically observed in both serum and whole blood and was also correlated with the results of the IFA. Conclusions:In conclusion, the CRNT for COVID-19 is a convenient assay system that can be performed in a BSL-2 laboratory with high specificity and sensitivity for evaluating the occurrence of neutralizing antibodies against SARS-CoV-2.

scholarly journals Recognition of Multiple Classes of Hepatitis C Antibodies Increases Detection Sensitivity in Oral Fluid

2001 ◽  
Vol 8 (6) ◽  
pp. 1267-1270 ◽  
Author(s):  
Jonathan F. Zmuda ◽  
Barbara Wagoneer ◽  
Lance Liotta ◽  
Gordon Whiteley
Keyword(s):  
Hepatitis C Virus ◽  
Hepatitis C ◽  
Oral Fluid ◽  
Detection Sensitivity ◽  
Secondary Antibody ◽  
Specific Detection ◽  
Serum Samples ◽  
Specificity And Sensitivity ◽  
Paired Serum ◽  
Antibody Cocktail

ABSTRACT Paired serum-oral fluid samples from 127 hepatitis C virus (HCV)-positive and 31 HCV-negative patients were tested for the presence of anti-HCV using the Ortho HCV 3.0 ELISA. Using the immunoglobulin G (IgG)-specific detection antibody provided with the HCV 3.0 ELISA we attained 100% sensitivity and specificity with serum samples; however, sensitivity in oral fluid samples was only 81%. By modifying the HCV 3.0 ELISA to utilize a secondary antibody cocktail that recognizes not only IgG but IgA and IgM as well, we attained 100% specificity and sensitivity with oral fluid samples.

scholarly journals Field Validation of the Use of RB51 as Antigen in a Complement Fixation Test To Identify Calves Vaccinated withBrucella abortus RB51

2001 ◽  
Vol 8 (2) ◽  
pp. 385-387 ◽  
Author(s):  
Rosanna Adone ◽  
Franco Ciuchini ◽  
Steven Olsen
Keyword(s):  
Complement Fixation ◽  
High Specificity ◽  
Antibody Responses ◽  
Test Results ◽  
Dot Blot ◽  
Serum Samples ◽  
Specificity And Sensitivity ◽  
Routine Surveillance ◽  
Dot Blot Assay ◽  
Smooth Strain

ABSTRACT In order to confirm the efficiency of an experimental RB51-based complement fixation (CF) test in identifying cattle vaccinated withBrucella abortus strain RB51, 831 sera from 110 vaccinated and 48 unvaccinated Hereford heifers of Iowa, collected for studies conducted in different years, were sent to Italy without coding to be tested in a CF test using RB51 as antigen. Most of the calves, aged from 3 to 10 months, were vaccinated subcutaneously with the recommended dosage of 1010 CFU of RB51 commercial vaccine, while only six calves received 109 CFU of the same vaccine. Serum samples for serologic testing, collected until 16 postinoculation weeks (PIW), were also tested by routine surveillance tests for brucellosis such as rose bengal plate and CF tests performed withB. abortus smooth strain 99 as control antigen. RB51 CF test results obtained by testing sera from cattle vaccinated in 1999 indicate that the sensitivity of the reaction is 97% at 2 to 3 PIW and 90% until 8 PIW and decreases to 65% at 12 PIW, the specificity remaining at 100%. Collectively, the results of this study confirm that serologic standard tests fail to detect antibodies to RB51 while the RB51-based CF test is able to monitor antibody responses to RB51 until 15 to 16 PIW with a specificity of 100%. In addition, unlike the RB51-based dot blot assay, which is the only test currently used to monitor antibody responses to RB51, the CF test also detected specific responses following vaccination with 109 CFU of RB51, although seroconversion was only 50% at 8 PIW. In conclusion, because of high specificity and sensitivity, the CF test described here can be used to efficaciously monitor serologic responses following RB51 vaccination in cattle and could also be employed to detect RB51 infection in humans exposed to this strain.

scholarly journals Cloning, expression and characterization of SeM protein of Streptococcus equi subsp. equi and evaluation of its use as antigen in an indirect ELISA

2014 ◽  
Vol 66 (4) ◽  
pp. 1015-1022 ◽  
Author(s):  
C.M. Moraes ◽  
F.R. Conceição ◽  
A.S.R. Rocha ◽  
A.G. Santos Júnior ◽  
L.M. Ribas ◽  
...  
Keyword(s):  
Immune Status ◽  
Indirect Elisa ◽  
High Specificity ◽  
Disease Diagnosis ◽  
Bacterial Isolation ◽  
Serum Samples ◽  
Gene Encoding ◽  
Specificity And Sensitivity ◽  
Streptococcus Equi

Strangles is an economically important horse disease caused by Streptococcus equi subsp. equi. The diagnosis can be confirmed either directly by bacterial isolation and PCR or by ELISA, which is an indirect method based on the detection of serum antibodies. The aim of this study was to clone, express and characterize the SeM protein of Streptococcus equi subsp. equi, evaluate its use as antigen in indirect ELISA and determine its performance to distinguish sera of negative, vaccinated and positive animals. This was initially performed by cloning the gene encoding the SeM protein and its expression in Escherichia coli. Subsequently, the protein produced was characterized and used as antigen in ELISA. Serum samples for evaluation were taken from 40 negative foals, 46 horses vaccinated with a commercial vaccine against strangles and 46 horses diagnosed with the disease. The test showed high specificity and sensitivity, allowing discrimination between negative and positive, positive and vaccinated animals, and vaccinated animals and negative sera. Thus, it was concluded that the protein produced rSeM, which can be used as antigen for disease diagnosis, and the described ELISA might be helpful to evaluate the immune status of the herd.

scholarly journals Lipopolysaccharide-Based Enzyme-Linked Immunosorbent Assay for Experimental Use in Detection of Antibodies to Lawsonia intracellularis in Pigs

2005 ◽  
Vol 12 (6) ◽  
pp. 693-699 ◽  
Author(s):  
J. J. Kroll ◽  
M. A. Eichmeyer ◽  
M. L. Schaeffer ◽  
S. McOrist ◽  
D. L. Harris ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Indirect Elisa ◽  
Experimental Studies ◽  
Antibody Test ◽  
Controlled Study ◽  
Future Research ◽  
Enzyme Linked Immunosorbent Assay ◽  
Lawsonia Intracellularis ◽  
Serum Samples ◽  
Specificity And Sensitivity

ABSTRACT An enzyme-linked immunosorbent assay (ELISA) for Lawsonia intracellularis was developed and compared with a whole-cell antigen-based immunofluorescence antibody test (IFAT). The antigen-containing lipopolysaccharide (LPS) was derived from Percoll gradient purified cultures of L. intracellularis by using a modification of the Westphal hot phenol procedure. The antigen was bound directly to polystyrene 96-well microtiter plates, and the assay was performed in an indirect ELISA format. Specificity and sensitivity values based on 80 known positive and 80 known negative serum samples from controlled experimental trials were 93.7% and 88.7%, respectively. Serological results from a controlled L. intracellularis challenge exposure study confirmed the high specificity and sensitivity of this assay (100% and 99.5%, respectively). Comparisons between the LPS ELISA and the IFAT in detecting anti-Lawsonia antibodies in this controlled study revealed significantly more LPS ELISA-positive pigs than IFAT-positive pigs on days 21, 28, 35, and 42 (P = 0.003, 0.030, 0.002, and 0.006, respectively). This indirect ELISA (LPS ELISA) test is an improved method of detecting antibodies in pigs soon after exposure to L. intracellularis, regardless of isolate type (vaccine or wild type) in experimental studies. The LPS ELISA may be used as a tool to support future research trials on vaccine efficacy and to further understand the immune response induced by L. intracellularis.

scholarly journals Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Hideki Tani ◽  
Miyuki Kimura ◽  
Long Tan ◽  
Yoshihiro Yoshida ◽  
Tatsuhiko Ozawa ◽  
...  
Keyword(s):  
Virus Infection ◽  
Whole Blood ◽  
Neutralizing Antibodies ◽  
High Specificity ◽  
Hospitalized Patients ◽  
Enzyme Linked Immunosorbent Assay ◽  
Pseudotyped Virus ◽  
Serum Samples ◽  
Live Virus ◽  
Specificity And Sensitivity

Abstract Background SARS-CoV-2 is a novel coronavirus that emerged in 2019 and is now classified in the genus Coronavirus with closely related SARS-CoV. SARS-CoV-2 is highly pathogenic in humans and is classified as a biosafety level (BSL)-3 pathogen, which makes manipulating it relatively difficult due to its infectious nature. Methods To circumvent the need for BSL-3 laboratories, an alternative assay was developed that avoids live virus and instead uses a recombinant VSV expressing luciferase and possesses the full length or truncated spike proteins of SARS-CoV-2. Furthermore, to measure SARS-CoV-2 neutralizing antibodies under BSL2 conditions, a chemiluminescence reduction neutralization test (CRNT) for SARS-CoV-2 was developed. The neutralization values of the serum samples collected from hospitalized patients with COVID-19 or SARS-CoV-2 PCR-negative donors against the pseudotyped virus infection evaluated by the CRNT were compared with antibody titers determined from an enzyme-linked immunosorbent assay (ELISA) or an immunofluorescence assay (IFA). Results The CRNT, which used whole blood collected from hospitalized patients with COVID-19, was also examined. As a result, the inhibition of pseudotyped virus infection was specifically observed in both serum and whole blood and was also correlated with the results of the IFA. Conclusions In conclusion, the CRNT for COVID-19 is a convenient assay system that can be performed in a BSL-2 laboratory with high specificity and sensitivity for evaluating the occurrence of neutralizing antibodies against SARS-CoV-2.

scholarly journals Evaluation of SARS-CoV-2 neutralizing antibodies using a vesicular stomatitis virus possessing SARS-CoV-2 spike protein

2020 ◽  
Author(s):  
Hideki Tani ◽  
Long Tan ◽  
Miyuki Kimura ◽  
Yoshihiro Yoshida ◽  
Hiroshi Yamada ◽  
...  
Keyword(s):  
Virus Infection ◽  
Whole Blood ◽  
Neutralizing Antibodies ◽  
High Specificity ◽  
Immunofluorescence Assay ◽  
Hospitalized Patients ◽  
Pseudotyped Virus ◽  
Serum Samples ◽  
Live Virus ◽  
Specificity And Sensitivity

AbstractSARS-CoV-2 is a novel coronavirus that emerged in 2019 and is now classified in the genus Coronavirus with closely related SARS-CoV. SARS-CoV-2 is highly pathogenic in humans and is classified as a biosafety level (BSL)-3 pathogen, which makes manipulating it relatively difficult due to its infectious nature. To circumvent the need for BSL-3 laboratories, an alternative assay was developed that avoids live virus and instead uses a recombinant VSV expressing luciferase and possesses the full length or truncated spike proteins of SARS-CoV-2. Furthermore, to measure SARS-CoV-2 neutralizing antibodies under BSL2 conditions, a chemiluminescence reduction neutralization test (CRNT) for SARS-CoV-2 was developed. The neutralization values of the serum samples collected from hospitalized patients with COVID-19 or SARS-CoV-2 PCR-negative donors against the pseudotyped virus infection evaluated by the CRNT were compared with antibody titers determined from an immunofluorescence assay (IFA). The CRNT, which used whole blood collected from hospitalized patients with COVID-19, was also examined. As a result, the inhibition of pseudotyped virus infection was specifically observed in both serum and whole blood and was also correlated with the results of the IFA. In conclusion, the CRNT for COVID-19 is a convenient assay system that can be performed in a BSL-2 laboratory with high specificity and sensitivity for evaluating the occurrence of neutralizing antibodies against SARS-CoV-2.

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