scholarly journals Development and Application of a Double-Antigen Sandwich Enzyme-Linked Immunosorbent Assay for Detection of Antibodies to Porcine Circovirus 2

2012 ◽  
Vol 19 (9) ◽  
pp. 1480-1486 ◽  
Author(s):  
Meng Ge ◽  
Wei Luo ◽  
Daliang Jiang ◽  
Runcheng Li ◽  
Wenwei Zhao ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Immunosorbent Assay ◽  
Sandwich Elisa ◽  
Test Procedure ◽  
Porcine Circovirus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Swine Industry ◽  
Elisa Kit ◽  
Porcine Circovirus 2

ABSTRACTA double-antigen sandwich enzyme-linked immunosorbent assay (ELISA) is described for detection of porcine circovirus 2 (PCV2) antibodies using the well-characterized recombinant PCV2 capsid protein. In a comparative test of 394 pig sera against an indirect immunofluorescence (IIF) test and a commercial ELISA kit (also based on the recombinant PCV2 capsid protein), the results showed that the diagnostic sensitivity, specificity, and accuracy of the assay were, respectively, 90.61, 94.02, and 91.62% compared with IIF and 94.38, 95.28, and 94.67% compared with the commercial ELISA kit. Assay of 12 PCV-free pigs over a 5-week period produced only PCV2-negative titers by all 3 methods. These results and the seroprofiles of 4 pig farms obtained by both the commercial ELISA kit and the double-antigen sandwich ELISA indicate that the sandwich ELISA is a reliable method for detection of antibodies to PCV2. Additionally, the method described here permits the use of undiluted test serum samples simultaneously loaded with horseradish peroxidase (HRP)-conjugated antigen into the test well, and the complete test procedure can be performed in less than 90 min. This double-antigen sandwich ELISA should be a useful tool to aid swine industry professionals in deciding the intervention strategies for the control of PCV2-associated diseases.

scholarly journals An indirect enzyme-linked immunosorbent assay using a recombinant truncated capsid protein of Porcine circovirus-2

2012 ◽  
Vol 24 (6) ◽  
pp. 1129-1132 ◽  
Author(s):  
Suphattra Jittimanee ◽  
Suparlark Nuntawan Na Ayudhya ◽  
Roongtham Kedkovid ◽  
Komkrich Teankum ◽  
Sanipa Suradhat ◽  
...  
Keyword(s):  
Capsid Protein ◽  
Immunosorbent Assay ◽  
Porcine Circovirus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Porcine Circovirus 2

A study of the correlation between Coxiella burnetii seropositivity and abortions in sheep in Eastern and Southeastern Turkey

2016 ◽  
Author(s):  
Ayse Kilic ◽  
Hakan Kalender
Keyword(s):  
Immunosorbent Assay ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Eastern Anatolia ◽  
Obligate Intracellular Bacterium ◽  
Serum Samples ◽  
Obligate Intracellular ◽  
Elisa Kit

Q fever is a zoonotic disease that occurs worldwide and is caused by the obligate intracellular bacterium Coxiella burnetii. Infected animals are usually asymptomatic, but infection can cause abortion and stillbirth in ruminants. The main purpose of this study was to evaluate prevalance of Coxiella burnetii infection in aborted and nonaborted sheep serum samples in Eastern Anatolia region by using enzyme-linked immunosorbent assay (ELISA). The determine of prevalance in sheep flocks from four provinces (Elazig, Malatya, Tunceli, Bitlis) and tested for anti-C.burnetii antibody detection, by means of Chekit Q fever Elisa kit. 350 serum samples obtained from flocks belonging aborted sheep showed that a total of 56 (16%) were detected seropositivity, whereas 171 serum samples obtained from nonaborted sheep flocks in 13 of the 171 (7.60%) for C.burnetii in seropositivity were observed. Coxiellosis should be considered an important cause of sheep with abortion history and nonaborted in Elazig and neighboring provinces.

scholarly journals Bovine Immunoglobulin E Levels of Bali Cattlesin Bangli and Nusa Penida island Bali Province, Indonesia

2019 ◽  
Vol 2 (2) ◽  
pp. 64
Author(s):  
Ni Ketut Suwiti ◽  
Luh Gde Surya Heryani ◽  
Desak Nyoman Dewi Indira Laksmi ◽  
Ni Nyoman Werdi Susari ◽  
I Nengah Kerta Besung ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Immunoglobulin E ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Commercial Enzyme ◽  
Elisa Kit ◽  
Bali Cattle ◽  
Bovine Immunoglobulin

The aim of this research was to detect identify levels of Bovine Immunoglobulin E (BoIg.E), can be used as an indicator of response immune in bali cattle.  Eighty serum samples were collected from Nusa Penida and Bangli region. Bovine Ig.E levels was measured using a commercial Enzyme Linked Immunosorbent Assay (ELISA) Kit. The data were analysis based on differences of farming characteristics andgeographic. The result of research that, of BoIg.E level of bali cattle kept in Bangli (34.16258 ?g/ml), was higher than Nusa Penida (22.26047 ?g/ml). We conclude that there was a significant effect of differences of farming characteristics and geographic conditions.

scholarly journals Development of a double monoclonal antibody–based sandwich enzyme-linked immunosorbent assay for detecting canine distemper virus

2020 ◽  
Vol 104 (24) ◽  
pp. 10725-10735
Author(s):  
Yuan Zhang ◽  
Gang Xu ◽  
Lu Zhang ◽  
Jiakai Zhao ◽  
Pinpin Ji ◽  
...  
Keyword(s):  
Monoclonal Antibody ◽  
Immunosorbent Assay ◽  
Colloidal Gold ◽  
Canine Distemper Virus ◽  
Sandwich Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Test Results ◽  
Canine Distemper ◽  
Serum Samples ◽  
Ideal Method

Abstract Canine distemper virus (CDV) infection causes mass mortality in diverse carnivore species. For effective virus surveillance, rapid and sensitive assays are needed to detect CDV in field samples. In this study, after BABL/c mice were immunized with recombinant CDV-fusion (F) protein, monoclonal antibodies (mAbs) against recombinant CDV-F protein (designated 1A5, 1A6, and 7D5) were produced using traditional hybridoma cell technology. Next, capture antibody (1A6, 800 ng/well) and horseradish peroxidase (HRP)–conjugated detection antibody (HRP-7D5, 1:100, 500 ng/well) were used in a double monoclonal antibody–based sandwich enzyme-linked immunosorbent assay (ELISA) for CDV detection after optimization of both mAb amounts per well using a checkerboard titration test. Based on sandwich ELISA test results for 120 known CDV-negative samples, the cutoff value for a positive result was set to an OD450 nm value ≥ 0.196. As compared with test results obtained from commercial immune colloidal gold test strips, the low limits of detection for the two assays were revealed to be 100 TCID50 per 100 μL. In addition, the sandwich ELISA agreed 100% and 96.4% with commercial immune colloidal gold test strips when testing serum and stool samples. The sandwich ELISA assay provided statistically similar CDV detection. Thus, the sandwich ELISA developed here to detect CDV in fecal and serum samples provided good sensitivity, high specificity, and good reproducibility and should serve as an ideal method for large-scale surveillance of CDV infections in carnivores. Key points • Three CDV mAbs that recognized different epitopes and bound to virion were generated. • The sandwich ELISA based mAbs to detect CDV in fecal and serum samples was developed. • The sandwich ELISA is an ideal method for detecting CDV infections in the field.

COMPARISON OF RADIAL IMMUNODIFFUSION AND SANDWICH-ELISA FOR THE MEASUREMENT IGG IN SERUM OF CALVES

2020 ◽  
pp. 27-28
Author(s):  
V. Klukina ◽  
O. Bogomolova ◽  
M. Romanenko ◽  
K. Tsar'kova
Keyword(s):  
Immunosorbent Assay ◽  
Sandwich Elisa ◽  
Test Procedure ◽  
Elisa Test ◽  
Radial Immunodiffusion ◽  
Passive Immunity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Single Radial Immunodiffusion

This investigation compared single radial immunodiffusion (sRID) and sandwich enzyme-linked immunosorbent assay (sandwich-ELISA) for the detection concentration of IgG in serum of calves. The results from each methods was shown positively correlated and that the ELISA test procedure would give more precise estimates of IgG concentration in serum for express diagnosis of failure of transfer of passive immunity of calves

scholarly journals ENZYME-LINKED IMMUNOSORBENT ASSAY DETECTION FOR MALIGNANCY USING ANTI-P53 ANTIBODIES

2021 ◽  
Vol 21 (1) ◽  
pp. 208-215
Author(s):  
MUHANNA ALSHAIBANI ◽  
Radin Maya Saphira Radin Mohamed ◽  
Ishak Mat ◽  
Adel AlGheethi ◽  
Jacinta Santhanam
Keyword(s):  
Immunosorbent Assay ◽  
P53 Protein ◽  
Malignant Tumour ◽  
Enzyme Linked Immunosorbent Assay ◽  
Malignant Tumours ◽  
Serum Samples ◽  
Discrimination Power ◽  
Elisa Kit ◽  
P53 Antibodies ◽  
Transformed Cell Lines

Tumour protein 53 (p53) plays an important role in the instruction of the cell cycle. In a variety of transformed cell lines, tumour protein is expressed in high amounts, and it is believed to contribute to transformation and malignancy. This research aimed to detect the anti-p53 antibodies in sera of patients with various malignant tumours and to evaluate the sensitivity and specificity of an enzyme-linked immunosorbent assay (ELISA). A case-control study was conducted on samples from 49 patients with various types of malignant tumours at Sultanah Bahiyah Hospital, Alor Setar, Kedah, Malaysia, and 32 healthy control cases with non‐malignant disease collected from Universiti Sains Malaysia clinic, Penang, Malaysia. The antibodies against p53 protein in the serum samples were analysed using the commercial ELISA kit, Calbiochem® p53- ELISAPLUS. The results showed that the rate of anti-p53 antibodies in patients with various malignant tumours was 13 out of 49 (26.5 %), compared with only 2 out of 32 (6.25%) in healthy controls (p < 0.001). The sensitivity of this kit reached 28.6% and the specificity was 93.8%. In conclusion, these results suggest that the anti-p53 antibodies can be detected in different sera of malignant tumour patients and the ELISA kit is highly specific; nevertheless, its discrimination power is not perfect because of its low sensitivity to determine the anti-p53 antibodies.

scholarly journals Autopsy-Controlled Prospective Evaluation of Serial Screening for Circulating Galactomannan by a Sandwich Enzyme-Linked Immunosorbent Assay for Hematological Patients at Risk for Invasive Aspergillosis

1999 ◽  
Vol 37 (10) ◽  
pp. 3223-3228 ◽  
Author(s):  
Johan Maertens ◽  
Jan Verhaegen ◽  
Hilde Demuynck ◽  
Penelope Brock ◽  
Gregor Verhoef ◽  
...  
Keyword(s):  
At Risk ◽  
Invasive Aspergillosis ◽  
Immunosorbent Assay ◽  
Predictive Value ◽  
Sandwich Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Hematological Patients ◽  
Increased Risk ◽  
Patients At Risk

Efforts to improve the diagnosis of invasive aspergillosis (IA) have been directed towards the detection of fungal antigens, including galactomannan (GM). However, previous evaluations of GM detection have been hampered by a lack of proven cases of IA and by a nonserial study design. This prospective study assessed the diagnostic value of serial screening for circulating GM by using a recently developed sandwich enzyme-linked immunosorbent assay (ELISA) for prolonged-neutropenic and/or steroid-treated patients with hematological disorders. Serum GM levels were monitored twice weekly for 186 consecutive patients at increased risk for IA. The patients were stratified according to the likelihood of IA (proven, probable, possible, and no evidence of IA) by using stringent criteria. Proven IA was defined by characteristic histopathological findings together with a positive culture forAspergillus species. Autopsy and culture from autopsy specimens was used to verify both positive and negative test results. A total of 2,172 serum samples were tested from 243 episodes (mean, 9 samples/episode). Based on the analysis of 71 patients with confirmed disease status (culture and histology), the sensitivity and specificity of serial GM monitoring were 92.6 and 95.4%, respectively. The positive predictive value was almost 93%, the negative predictive value was 95%, and the efficacy was 94%. False-positive reactions occurred at a rate of nearly 8%, although this figure might have been overestimated. Less than 1% of all tested sera were considered inconclusive. In more than half of the cases, antigenemia was detected before clinical suspicion of IA (median, 6 days before). Serial determination of serum GM by the sandwich ELISA technique is a sensitive tool for the diagnosis of IA in hematological patients at risk. This approach may substantially influence clinical management with regard to preemptive and empirical antifungal therapy.

Can Osteopontin Be Considered a Biomarker for Endometriosis?

2017 ◽  
Vol 68 (9) ◽  
pp. 2132-2134
Author(s):  
Daniela Roxana Albu (Matasariu) ◽  
Elena Mihalceanu ◽  
Alina Pangal ◽  
Carmen Vulpoi ◽  
Mircea Onofriescu ◽  
...  
Keyword(s):  
Serum Level ◽  
Immunosorbent Assay ◽  
Pelvic Pain ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Multifactorial Disease ◽  
Elisa Kit ◽  
Healthy Women ◽  
Progesterone Treatment ◽  
Large Dispersion

Endometriosis is a multifactorial disease that is manifested by infertility and pelvic pain. The purpose of the study was to evaluate the effect of progesterone treatment on the serum level of osteopontin, a multipotent cytokine, in patients with endometriosis. The study was prospective and we evaluated osteopontin levels that were measured in the serum of 40 patients with endometriosis and 12 healthy women using a standardized Enzyme-Linked Immunosorbent Assay (ELISA) kit. Osteopontin seric levels were lower in endometriosis patients and increased after progesterone treatment. Because of the large dispersion of data even in the control group, we find the association between osteopontin and endometriosis questionable.

scholarly journals Exploratory field study on the effects of porcine circovirus 2 (PCV-2) sow vaccination at different physiological stages mimicking blanket vaccination

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Patricia Pleguezuelos ◽  
Marina Sibila ◽  
Raúl Cuadrado ◽  
Rosa López-Jiménez ◽  
Diego Pérez ◽  
...  
Keyword(s):  
Quantitative Polymerase Chain Reaction ◽  
Negative Control Group ◽  
Negative Control ◽  
Late Gestation ◽  
Antibody Detection ◽  
Porcine Circovirus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Group ◽  
Physiological Status ◽  
Porcine Circovirus 2

Abstract Background The objective of the present study was to explore the benefits of Porcine circovirus 2 (PCV-2) blanket vaccination in a sow herd on productive parameters, PCV-2 infection and immune status in sows and their progeny. For this purpose, 288 sows were distributed among four balanced experimental groups. One group remained as negative control group and the other three received 1 mL of PCV-2 Ingelvac Circoflex® intramuscularly at different productive cycle moments: before mating, mid gestation (42–49 days post-insemination) or late gestation (86–93 days post-insemination); phosphate buffered saline (PBS) was used as negative control item. Reproductive parameters from sows during gestation and body weight of their progeny from birth to weaning were recorded. Additionally, blood was collected from sows at each vaccination time and piglets at 3 weeks of age. Moreover, up to 4 placental umbilical cords (PUC) per sow were taken at peri-partum. Sera from sows and piglets were analysed for PCV-2 antibody detection using an enzyme-linked immunosorbent assay (ELISA). Sera from sows and PUC were tested to quantify viraemia using a real time quantitative polymerase chain reaction (qPCR) assay. Results Globally, results indicated that vaccinated sows showed heavier piglets at birth and at weaning, less cross-fostered piglets, lower viral load at farrowing as well as in PUC, and higher antibody levels at farrowing, compared to non-vaccinated ones. When all groups were compared among them, sows vaccinated at mid or late gestation had heavier piglets at birth than non-vaccinated sows, and lower proportion of PCV-2 positive PUC. Also, cross-fostering was less frequently practiced in sows vaccinated at pre-mating or mid gestation compared to non-vaccinated ones. Conclusions In conclusion, the present study points out that PCV-2 sow vaccination at different time points of their physiological status (mimicking blanket vaccination) offers benefits at production and serological and virological levels.

Sign in / Sign up

Export Citation Format

Share Document