scholarly journals ELISA test metapneumoviral infection of bird: methodology of development and use in veterinary practice

2021 ◽  
pp. 24-31
Author(s):  
O.V. Tsinoviy ◽  
◽  
L.I. Nalyvayko ◽  
Keyword(s):  
Test System ◽  
Elisa Test ◽  
High Price ◽  
Enzyme Linked Immunosorbent Assay ◽  
Nucleotide Sequencing ◽  
Test Systems ◽  
Subtype B ◽  
Antibody Titers ◽  
Scientific Documentation ◽  
And Control

In the absence of diagnostic kits for the detection of antibodies to metapneumovirus infection (MPVI) epizootological monitoring in Ukrainian farms is practically not carried out, imported test systems have a "sky-high" price, so there is a need for domestic methods of diagnosing this disease. The most accurate, easy-to-use method is ELISA-based test systems (enzyme-linked immunosorbent assay). A diagnostic ELISA test system for the detection of antibodies to MPVI has been developed and it has been established that this diagnosticum should be used in the practice of veterinary medicine for serological control of metaviral virus infection. The optimal ratios of components for the manufacture of ELISA test system have been worked out. The form of calculation of antibody titers in blood sera of chickens when testing them in one dilution is calculated. The sensitivity and specificity of the ELISA test system were determined (comparative analysis of serum testing results in ELISA, RNGA and RN). Scientific documentation has been developed – instructions for the manufacture and control of ELISA test systems for the detection of antibodies to metapneumovirus infection in the serum of chickens and instructions for its use. Indication and identification of the obtained virus isolate was carried out using polymerase chain reaction (PCR) and nucleotide sequencing. Based on the studies carried out in the suspension of the internal organs of turkeys (trachea, lungs), a virus belonging to subtype B of the genus Metapneumovirus, subfamily Pneumovirinae, family Paramyxoviridae of the order Mononegavirales was revealed. By comparing the nucleotide sequence of the G gene fragment of the PVT-09/B strain with the sequences of strains and isolates of the avian metapneumovirus subtype B published in the GenBank database, it was found that the metapneumovirus isolated from sick turkeys is phylogenetically close to the Brazilian strains 27A-07 2007 and MPV/B/Brazil-07/USP-08 G

scholarly journals Comparative Analysis of the Analytical Sensitivity of ELISA Test System DIA®-SARS-CoV-2-Ag-R with Rapid Tests for Viral Antigen SARS-CoV-2 Detection

2021 ◽  
Vol 83 (3) ◽  
pp. 66-71
Author(s):  
А.Y. Horlov ◽  
◽  
V.H. Serdiuk ◽  
О.K. Kiselova ◽  
A.O. Shevchuk ◽  
...  
Keyword(s):  
Viral Antigen ◽  
False Negative ◽  
Medical Science ◽  
Rapid Test ◽  
Test System ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Analytical Sensitivity ◽  
Rapid Tests ◽  
Positive Results

A novel coronavirus severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease (COVID)-19, that emerged as a major pandemic. SARS-CoV-2 was identified as a betacoronavirus. Nucleocapsid protein (NP) is evolutionary high protein homologies and solid structure protein for SARS-CoV-2 detection as opposed to other proteins, that aren`t reliable as a single viral antigen during diagnostics methods. The viral RNA can be detected from nasal and pharyngeal swabs and bronchoalveolar lavage samples by PCR assay. However, the wrong collection of samples can lead to false-negative diagnosis and have dangerous consequences at this stage of pandemic. One of efficient and accurate methodological approaches for the screening of pathogens are serological assays. Aim. Evaluation and comparison of the sensitivity of invented DIA®-SARS-CoV-2-Ag-R enzyme-linked immunosorbent assay (ELISA)-based test system and commercial rapid tests, which detect the viral antigen of SARS-CoV-2. Methods. We carried out a comparison of DIA®-SARS-CoV-2-Ag-R and existed commercial test systems, which are used to detect the antigen of SARS-CoV-2 virus. Rapid tests are intended for nasopharyngeal swabs, but we proposed a protein of our own manufacture – recombinant NP as a calibrator. The detection limit was calibrated by standard CFAR #100982 NIBSC, UK. We had determined levels of NP (1400, 900, 750, 640 and 280 pg/mL) that we used as a sample for the rapid tests. The COVID-19 Ag Rapid Tests were performed according to the manufactures instructions at room temperature. Results. DIA®-SARS-CoV-2-Ag-R detected 10 pg/mL of in-house standard of recombinant SARS-CoV-2 NP. The positive results were observed using 1400, 900, 750 pg/mL, while 640 and 280 pg/mL samples were performed as negative in ABBOTT-PanBio test. The rapid tests manufactured by МBU, BIOTIME, Core Technology, SD BIOSENSOR and Turklab showed positive results only in 1400 pg/mL concentration. NP of lower levels was detected as a negative sample. The LEPU MEDICAL test was evaluated as positive sample using 900 pg/mL. The rapid test manufactured by Green Cross Medical Science Corp. showed negative results for all levels of NP. It can mean that the sensitivity of test is lower and demands higher level of antigen to detect the presence of SARS-CoV-2. Conclusions. The study presented an excellent analytical sensitivity of DIA®-SARS-CoV-2-Ag-R against commercial Antigen rapid tests. Thus, invented ELISA test system can be recommended for widespread usage for detection and confirmation of acute stage of SARS-CoV-2 infection.

scholarly journals Development of new immunoanalytical test systems for diagnostics of potato blackleg caused by Dickeya spp. bacteria

2021 ◽  
Vol 16 (3) ◽  
pp. 198-214
Author(s):  
Shyatesa Razo ◽  
Pavel A. Galushka ◽  
Yuri A. Varitsev ◽  
Anatoly V. Zherdev ◽  
Irina V. Safenkova ◽  
...  
Keyword(s):  
Detection Limit ◽  
Polyclonal Antibodies ◽  
Test System ◽  
Diagnostic Tools ◽  
Enzyme Linked Immunosorbent Assay ◽  
Potato Seed ◽  
Potato Blackleg ◽  
Bacterial Diseases ◽  
High Affinity ◽  
Test Systems

Potato blackleg caused by Dickeya spp. bacteria is one of the most important bacterial diseases of potatoes. The rapid spread of this disease in the territory of Russia requires new effective diagnostic tools for the timely detection of infection. To solve this problem, antisera specific to Dickeya spp. were obtained. Polyclonal antibodies isolated from antisera have shown high affinity for the main species of Dickeya spp. ( D. solani, D. dianthicola, D. chrysanthemi, D. dadantii, D. paradisiaca ). Enzyme linked immunosorbent assay (ELISA) and lateral flow immunoassay (LFIA) test systems have been developed based on specific and high affinity antibodies that were obtained. For ELISA, the detection limit was 0.8 105 cells/mL for D. solani and 2 104 cells/mL for D. dianthicola . For LFIA, suitable for use in non-laboratory conditions, the detection limit of D. solani was 2 105 cells/mL and the analysis time was 15 minutes. When testing potato seed material, LFIA test system confirmed positive results of ELISA determination in 75 % of samples, and negative - in 100 % of samples.

Establishment of a Flexible Platform for an Automated Brain-Derived Neurotrophic Factor—ELISA

2007 ◽  
Vol 12 (4) ◽  
pp. 219-229
Author(s):  
Ilka Schneider ◽  
Paul Stoll ◽  
Daniel Haller ◽  
Kerstin Thurow
Keyword(s):  
Neurotrophic Factor ◽  
Brain Derived Neurotrophic Factor ◽  
Sample Volume ◽  
Test System ◽  
Elisa Test ◽  
Small Sample ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Automated Processing ◽  
High Flexibility

Enzyme-linked immunosorbent assay (ELISA) is a standard immunoassay to estimate protein concentrations in a variety of samples. An automated ELISA procedure was established that allows the measurement of the concentration of numerous proteins in a small sample volume. The automated ELISA procedure was tested by measuring the concentration of brain-derived neurotrophic factor in serum samples from humans. It was shown that the reproducibility of protein concentrations in three assay plates within one automated run was greater than 92%, within independent automated experiments greater than 96%. Automated processing with a maximum capacity of six plates and 240 serum samples per run showed an average interassay precision of 91 %. Serial dilutions of randomly chosen sera displayed significant linearity of recovered concentration. In conclusion it can be stated that the automated assay provides high flexibility with highly reproducible results and has several advantages compared to the manual method, including less operator dependence and faster sample throughput. The investigated ELISA test system seems to exhibit analytical characteristics indicating further clinical application. (JALA 2007;12:219–29)

scholarly journals ELISA Based Anthrax Antibody Titer in Cattle Induced by Locally Prepared Anthrax Vaccine Originated from Sterne F-24 Strain in Bangladesh

2016 ◽  
Vol 4 (1) ◽  
pp. 36-38 ◽  
Author(s):  
Jayedul Hassan ◽  
Md Bahanur Rahman ◽  
Shah Md Ziqrul Haq Chowdhury ◽  
Shushanto Kumar Rabidas ◽  
Md Shafiullah Parvej ◽  
...  
Keyword(s):  
Reference Value ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Post Vaccination ◽  
Blood Samples ◽  
Anthrax Vaccine ◽  
Antibody Titers ◽  
Antibody Levels ◽  
And Control ◽  
Adequate Antibody Response

Vaccination is usually practiced to prevent and control anthrax in Bangladesh. For this purpose, vaccine prepared from Sterne F-24 strain ofBacillus anthracisby Livestock Research Institute (LRI), Mohakhali, Dhakahas long been used in this country. However, in some cases anthrax occurred in vaccinated animals in Bangladesh. A total of 100 cattle at LalTeer Livestock Research and Development Farm, LalTeerLivestock Limited, Bangladesh, aging between 3-6 years and weighing between 250-400 kg were randomly selected for vaccination purpose. Blood samples (n=100) were collected before the vaccination for collecting pre-vaccination serum, andthe animals were vaccinated (at 1 mL/animal; 1x107 spores/mL) with the anthrax vaccine produced by LRI. All blood samples from the vaccinated animals were collected on day 7, 28, 60, 90, 120, 150, 180, 240, 270, 300, 330, and 360 of post-vaccination, and serum samples were prepared. The antibody levels in the serum samples against anthrax were monitored using an Enzyme-Linked Immunosorbent Assay (ELISA). Over the course of 12 months, the antibody titers were found at the level higher than the reference value. Though there were reports on anthrax suspected cases in this farm, no such cases were reported during the study period. Thus, the vaccine appears to induce adequate antibody response against anthrax in Bangladesh.Microbes and Health, January 2015. 4(1): 36-38

scholarly journals STUDY OF ELISA TEST-SYSTEMS OF DIFFERENT FORMATS FOR DETECTION OF MEASLES VIRUS SPECIFIC IgM IN DIFFERENT GEOGRAPHIC ZONES

2018 ◽  
Vol 8 (2) ◽  
pp. 230-234
Author(s):  
M. A. Bichurina ◽  
N. V. Zheleznova ◽  
I. N. Lavrentieva ◽  
A. Yu. Antipova ◽  
L. B. Kulyashova ◽  
...  
Keyword(s):  
Blood Serum ◽  
False Positive ◽  
Measles Virus ◽  
Test System ◽  
Elisa Test ◽  
Healthy Adults ◽  
Igg Antibodies ◽  
Serum Samples ◽  
Test Systems ◽  
Igm Antibodies

Detection of the measles virus (MV) specific IgM antibodies in blood serum of patients is considered to be the main standard for the  laboratory confirmation of measles diagnosis, the test being  acknowledged by WHO. As it was demonstrated earlier the specific  IgM antibodies as the marker of the acute MV infection were  detected in 97.2–100% of blood serum samples from patients using  the ELISA test-systems of the “capture” format (Microimmun Ltd.  and Vector Best). In case when the ELISA test-system of the  “indirect” format (Siemens, Germany) was used only 63.9% of these sera turned to be IgM positive. And on the contrary using the  “indirect” format ELISA test-system Euroimmun, Germany, for  detection of the MV specific IgM the false positive results were  obtained.The aim of the study was the comparative evaluation of  the different format ELISA test-systems used for the detection of the  MV specific IgM antibodies in blood sera of patients and healthy  adults collected in different geographic zones.Materials and  methods. In total 108 serum specimens collected in 2015–2017 were studied: from healthy adult Guineans, residents of the Republic of Guinea (RG); patients aged 1–70 with the initial  “infectious mononucleosis”, “infectious cytomegalovirus” and  “rubella” diagnosis and taken from the bank of sera in the  Subnational Measles/Rubella laboratory, StP Measles/Rubella RC in  NWFR. The MV specific IgM antibodies were detected using the commercial ELISA test-systems “VectoMeasles-IgM” (Vector-Best,  Russia) (“capture” format) and “Anti-Measles Virus ELISA IgM (NP)”  (Euroimmun Medizinische Labordiagnostik AG, Germany) («indirect»  format). The specific Epshtein-Barr Virus (EBV) IgM and IgG  antibodies were detected with the commercial ELISA test-systems «DS-ELISA-anti-EBV-VCA-M», «DS-ELISA-anti-EBV-EA-G» and «DS-ELISA-anti-EBV-NA-G» (“Diagnostic Systems”, Russia).Results and discussion. The MV specific IgM antibodies were not revealed in the total of 108 blood serum samples from the healthy adults and patients, residents of the Russia and of the RG, with the “capture”  format “VectoMeasles-IgM” ELISA test-system. The absence of the  acute MV infection was also confirmed by the high measles immunity  level (i.e. IgG MV antibodies titers) as well as by detection of the IgG antibodies of high avidity. At the same time in 6  of 108 total sera (5.5%) IgM MV antibodies were detected with the «indirect» format ELISA test system Euroimmun, Germany. In these 6 sera the EBV specific antibodies were also evidenced. The results  obtained demonstrate the nonspecific reaction due to the possible  reactivity with anti-EBV antibodies. Besides this the different  percentage of the false positive reactions in sera from healthy  adults, residents of the RG and residents of Russia was determined  — 8.5±4.0% and 3.2±2.2% correspondently. Thus the preliminary  results, and to get the final results for general conclusions increase  of the total amount of the clinical specimens under studying is of extremely importance.

scholarly journals Development and Evaluation of an Immunochromatographic Strip for Detection of Streptococcus Suis Type 2 Antibody

2007 ◽  
Vol 19 (4) ◽  
pp. 355-361 ◽  
Author(s):  
Junxing Yang ◽  
Meilin Jin ◽  
Jianfeng Chen ◽  
Ying Yang ◽  
Pei Zheng ◽  
...  
Keyword(s):  
Capsular Polysaccharide ◽  
Protein A ◽  
Streptococcus Suis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Staphylococcal Protein A ◽  
Immunochromatographic Strip ◽  
Specificity And Sensitivity ◽  
Antibody Titers ◽  
Serological Surveillance ◽  
And Control

In this study, an immunochromatographic strip (ICS) was developed for the detection of antibody against Streptococcus suis serotype 2 (SS2). Colloidal gold particles labeled with staphylococcal protein A (SPA), which can bind to the FC fragment of mammalian immunoglobulin, were used as the detector reagent. The capsular polysaccharide (CPS) of SS2 and affinity-purified IgG from a healthy naive pig were immobilized on test and control regions of a nitrocellulose membrane, respectively. The ICS was used to 1) detect anti-CPS antibody in 14 sera taken from 4 SS2-infected pigs, 24 sera from pigs hyperimmunized with SS2, and 68 sera from pigs inoculated or infected with bacteria other than SS2; 2) determine anti-CPS antibody titers of 20 positive sera for comparison with enzyme-linked immunosorbent assay (ELISA); and 3) detect anti-CPS antibody in 226 clinical sera taken from diseased pigs also for comparison with ELISA. An ELISA used as a reference test determined the specificity and sensitivity of the ICS to be 97.1% and 86.3%, respectively. There was excellent agreement between the results obtained by ELISA and the ICS (kappa = 0.843). Additionally, there was strong agreement between the results of bacterial isolation from pig tonsils and ICS test (kappa = 0.658). Because it is rapid and easy to use, the test is suitable for the serological surveillance of SS2 at farms.

scholarly journals Post-vaccinal reaction for some vaccines used against Newcastle disease in Sulaimaniyah province

2012 ◽  
Vol 11 (1) ◽  
pp. 133
Author(s):  
E.A. Abdul Ahad
Keyword(s):  
Newcastle Disease ◽  
Significant Variation ◽  
Histopathological Examination ◽  
Elisa Test ◽  
Bursa Of Fabricius ◽  
Control Group ◽  
Broiler Chicks ◽  
Treatment Groups ◽  
Antibody Titers ◽  
And Control

This study was conducted to investigate the safety of the most commonly used Newcastle disease vaccines in Al-Sulaimaniyah province (LaSota and Clone30). A total of 225 one-day-old broiler chicks of Ross 308 breed were investigated for their maternal-derived antibody (MDA) titers by ELISA test. Subsequently, these chicks were divided randomly into 3 equal groups; ( 2 treatment groups, a T1 group which was vaccinated by LaSota vaccine and T2 group which was vaccinated by the Clone30 vaccine). and control non-vaccinated group. ELISA test was used to investigate the antibody titers against NDV in all groups on day 10 post second vaccination at 34 days of chicks age and tissue biopsies were obtained for histopathological examination from the trachea, spleen, Bursa of Fabricius and thymus to explore the tissue changes that may be induced by the vaccine. Significant variation was observed in the means of antibody titers against NDV between the control and treatment groups, whereas, no significant variation was observed between the treatment groups themselves. The histopathological examination results showed that a reactive lymphocytic response was observed in both treatment groups compared to the control group. In addition, focal epithelial sloughing and mucopurulent exudates were observed in the trachea of T1 group chicks only. The result of this study showed that the NDV vaccine of clone30 is approximate of the same efficiency and more secure than LaSota vaccine.

Application of commercial test-systems to identify gram-negative facultatively anaerobic bacteria

2016 ◽  
Vol 3 (1) ◽  
pp. 43-48 ◽  
Author(s):  
V. Patyka ◽  
L. Butsenko ◽  
L. Pasichnyk
Keyword(s):  
Erwinia Amylovora ◽  
Anaerobic Bacteria ◽  
Biochemical Properties ◽  
Test System ◽  
Phytopathogenic Bacteria ◽  
Gram Negative ◽  
Test Systems ◽  
Facultatively Anaerobic ◽  
Microbiological Methods

Aim. To validate the suitability of commercial API 20E test-system (bioMerieux) for the identifi cation and characterization of facultative gram-negative phytopathogenic bacterial isolates. Methods. Conventional mi- crobiological methods, API 20E test-system (bioMerieux) according to the manufacturer’s instructions. Re- sults. The identifi cation results for Erwinia amylovora, Pectobacterium carotovorum and Pantoea agglome- rans isolates were derived from the conventional and API 20E test systems, which, were in line with the literature data for these species. The API 20E test-system showed high suitability for P. agglomerans isolates identifi cation. Although not all the species of facultatively anaerobic phytopathogenic bacteria may be identi- fi ed using API 20E test-system, its application will surely allow obtaining reliable data about their physiologi- cal and biochemical properties, valuable for identifi cation of bacteria, in the course of 24 h. Conclusions. The results of tests, obtained for investigated species while using API 20E test-system, and those of conventional microbiological methods coincided. The application of API 20E test-system (bioMerieux) ensures fast obtain- ing of important data, which may be used to identify phytopathogenic bacteria of Erwinia, Pectobacterium, Pantoea genera.

scholarly journals Anti-HPV16 Antibody Titers Prior to an Incident Cervical HPV16/31 Infection

Viruses ◽  
2021 ◽  
Vol 13 (8) ◽  
pp. 1548
Author(s):  
Ana Gradissimo ◽  
Viswanathan Shankar ◽  
Fanua Wiek ◽  
Lauren St. Peter ◽  
Yevgeniy Studentsov ◽  
...  
Keyword(s):  
High Risk ◽  
Hpv Vaccine ◽  
Enzyme Linked Immunosorbent Assay ◽  
Incident Detection ◽  
Antibody Titers ◽  
Circulating Antibodies ◽  
Prospective Association ◽  
Nested Case Control ◽  
Incident Cases

The goal of this study was to investigate the serological titers of circulating antibodies against human papillomavirus (HPV) type 16 (anti-HPV16) prior to the detection of an incident HPV16 or HPV31 infection amongst vaccinated participants. Patients were selected from a prospective post-HPV vaccine longitudinal cohort at Mount Sinai Adolescent Health Center in Manhattan, NY. We performed a nested case–control study of 43 cases with incident detection of cervical HPV16 (n = 26) or HPV31 (n = 17) DNA who had completed the full set of immunizations of the quadrivalent HPV vaccine (4vHPV). Two control individuals whom had received three doses of the vaccine (HPV16/31-negative) were selected per case, matched on age at the first dose of vaccination and follow-up time in the study: a random control, and a high-risk control that was in the upper quartile of a sexual risk behavior score. We conducted an enzyme-linked immunosorbent assay (ELISA) for the detection of immunoglobulin G (IgG) antibodies specific to anti-HPV16 virus-like particles (VLPs). The results suggest that the average log antibody titers were higher among high-risk controls than the HPV16/31 incident cases and the randomly selected controls. We show a prospective association between anti-HPV16 VLP titers and the acquisition of an HPV16/31 incident infection post-receiving three doses of 4vHPV vaccine.

scholarly journals Validation of a Novel Commercial ELISA Test for the Detection of Antibodies against Coxiella burnetii

Pathogens ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1075
Author(s):  
Salvatore Ledda ◽  
Cinzia Santucciu ◽  
Valentina Chisu ◽  
Giovanna Masala
Keyword(s):  
Diagnostic Accuracy ◽  
Phase Ii ◽  
Coxiella Burnetii ◽  
Q Fever ◽  
Animal Health ◽  
Elisa Test ◽  
Clinical Diagnostics ◽  
Complex Life Cycle ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specificity And Sensitivity

Q fever is a zoonosis caused by Coxiella burnetii, a Gram-negative pathogen with a complex life cycle and a high impact on public and animal health all over the world. The symptoms are indistinguishable from those belonging to other diseases, and the disease could be symptomless. For these reasons, reliable laboratory tests are essential for an accurate diagnosis. The aim of this study was to validate a novel enzyme-linked immunosorbent assay (ELISA) test, named the Chorus Q Fever Phase II IgG and IgM Kit (DIESSE, Diagnostica Senese S.p.A), which is performed by an instrument named Chorus, a new device in medical diagnostics. This diagnostic test is employed for the detection of antibodies against C. burnetii Phase II antigens in acute disease. Our validation protocol was performed according to the Italian Accreditation Body (ACCREDIA) (Regulation UNI CEI EN ISO/IEC 17025:2018 and 17043:2010), OIE (World Organization for Animal Health), and Statement for Reporting Studies of Diagnostic Accuracy (STARD). Operator performance was evaluated along with the analytical specificity and sensitivity (ASp and ASe) and diagnostic accuracy of the kit, with parameters such as diagnostic specificity and sensitivity (DSp and DSe) and positive and negative predictive values (PPV and NPV), in addition to the repeatability. According to the evaluated parameters, the diagnostic ELISA test was shown to be suitable for validation and commercialization as a screening method in human sera and a valid support for clinical diagnostics.

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