scholarly journals Variations in the detection of anti-PEDV antibodies in serum samples using three diagnostic tests – short communication

2018 ◽  
Vol 66 (2) ◽  
pp. 337-342 ◽  
Author(s):  
Jan Plut ◽  
Ivan Toplak ◽  
Marina Štukelj

Over the last few years several porcine epidemic diarrhoea (PED) outbreaks have been discovered in Europe including the first PED case in Slovenia in January 2015. The aim of this study was to determine when PED virus (PEDV) infection started in Slovenia. Serum samples collected between 2012 and 2016 were tested. Three hundred and seventy-five serum samples were collected from 132 Slovenian small, one-site pig farms. Samples were tested for PEDV antibodies utilising three different serological methods: commercially-available indirect ELISA, in-house blocking ELISA test and Immunoperoxidase Monolayer Assay (IPMA) test. One hundred and seventy (45.33%) tested samples were found positive by the commercially-available ELISA test kit, and 10 (5.68%) of these 170 samples found positive were positive by the in-house blocking ELISA. Only these 10 samples were collected from a farm where clinical signs of PED infection had been observed and PEDV was confirmed by RT-PCR methodology; the other 160 samples were collected randomly. Thirty-two samples with the highest S/P value obtained with the commercial ELISA were all negative with IPMA. Reasons for the high variance in the results obtained remain unclear; more research is required to ensure higher sensitivity and specificity in terms of PEDV antibody tests and other PED diagnostic methods.

Author(s):  
E.V. Molchanova ◽  
D.N. Luchinin ◽  
A.O. Negodenko ◽  
D.R. Prilepskaya ◽  
N.V. Boroday ◽  
...  

The paper presents data from the monitoring studies’ results of arbovirus infections transmitted by mosquitoes in the Volgograd region. West Nile virus antigen (WNV) in 9 samples, Tahyna virus in one sample, Batai virus in two samples were detected in the study of 110 samples of field material (blood-sucking mosquitoes) by ELISA test. Antibodies to WNV in 16.58 percent of the samples, to tick-borne encephalitis virus in 1.08 percent, to viruses of the California serogroup and Ukuniemi in 1.09 percent, to the virus Sindbis in 2.17 percent were detected as a result of the study of blood serum samples from donors in the Volgograd region. Thus, we obtained data on the probable presence of the Batai, Sindbis, Ukuniemi and Californian serogroup viruses along with the circulation of WNV on the territory of the Volgograd region.


2019 ◽  
Vol 17 (1) ◽  
Author(s):  
Zhiqian Ma ◽  
Tianyu Wang ◽  
Zhiwei Li ◽  
Xuyang Guo ◽  
Yangsheng Tian ◽  
...  

Abstract Background Porcine epidemic diarrhea virus (PEDV), which is characterized by severe watery diarrhea, vomiting, dehydration and a high mortality rate in piglets, leads to enormous economic losses to the pork industry and remains a large challenge worldwide. Thus, a rapid and reliable method is required for epidemiological investigations and to evaluate the effect of immunization. However, the current diagnostic methods for PEDV are time-consuming and very expensive and rarely meet the requirements for clinical application. Nanobodies have been used in the clinic to overcome these problems because of the advantages of their easy expression and high level of stability. In the present work, a novel biotinylated nanobody-based blocking ELISA (bELISA) was developed to detect anti-PEDV antibodies in clinical pig serum. Results Using phage display technology and periplasmic extraction ELISA (PE-ELISA), anti-PEDV N protein nanobodies from three strains of PEDV were successfully isolated after three consecutive rounds of bio-panning from a high quality phage display VHH library. Then, purified Nb2-Avi-tag fusion protein was biotinylated in vitro. A novel bELISA was subsequently developed for the first time with biotinylated Nb2. The cutoff value for bELISA was 29.27%. One hundred and fifty clinical serum samples were tested by both newly developed bELISA and commercial kits. The sensitivity and specificity of bELISA were 100% and 93.18%, respectively, and the coincidence rate between the two methods was 94%. Conclusions In brief, bELISA is a rapid, low-cost, reliable and useful nanobody-based tool for the serological evaluation of current PEDV vaccines efficacy and indirect diagnosis of PEDV infection.


Author(s):  
Flywell Kawonga ◽  
Gerald Misinzo ◽  
Dylo Pemba ◽  
Leonard Mboera ◽  
Isaac Thom Shawa

Chikungunya is a mosquito-borne viral disease caused by Chikungunya virus (CHIKV. We conducted this study determine the seroprevalence and clinical presentation of Chikungunya infection among outpatients seeking healthcare in Mzuzu City, Malawi. Blood samples were collected from malaria negative and non-septic febrile outpatients with fevers ≥38 °C, for not more than 5 days. The enzyme- linked immunosorbent assay (ELISA) test was used to detect anti-CHIKV IgM antibodies and its results were used to determine seroprevalence of Chikungunya. A total of 119 serum samples were tested, of these, 73 (61.3%) tested positive for anti-CHIKV IgM antibodies by ELISA. Laboratory requisition forms were used to capture demographic information such as age, sex, clinical signs and symptoms presented by the enrolled patients. Age groups of 1-9, 10- 19, 20- 29, 30- 39, 40- 49, and ≥50 years had 17.8% (n= 13), 12.3 %,( n=9), 15.1%) (n=11), 19.2%; (n=14), 17.8% (n=13) and 17.8% (n=13) proportion of seroprevalence respectively. Most of the CHIKV infected individuals presented with fever (52.05%), joint pain (45.21%) and abdominal pain (42.67%). The presence of anti- CHIKV IgM antibodies suggest the presence of recent CHIKV infection and therefore accurate laboratory assays are highly recommended for CHIKV diagnosis and appropriate management of febrile patients.


2019 ◽  
Author(s):  
Sadik Akgun ◽  
Sezgin Barutcu ◽  
Sumeyya Capuk ◽  
Arzu Tanriverdi ◽  
Serihan Kübra Emikoglu ◽  
...  

Abstract Background: Helicobacter pylori (H. pylori) is a significant contributor of various gastrointestinal disorders and cancers all around the world. Its diagnosis is dependent on several qualitative and quantitative methods. The present study aims to compare the results of rapid cassette and micro ELISA test methods for diagnosis of H. pylori and determining associations with patient endoscopy reports. Methods: The study was performed using blood samples collected from 224 patients (142 (63%) females and 82 (37%) males) in various clinics between January 2018 and August 2019, which were sent to the Clinical Microbiology Laboratory of Training Hospital. Serum samples obtained after centrifugation of the blood samples were initially tested with rapid H. pylori IgG cassette method, and afterwards in the auto analyzer using ELISA assays specific for H. pylori. Results: Upper gastrointestinal system endoscopy was performed in 88 of these patients, and biopsy results confirmed definitive diagnosis of H. pylori infection in 63 of the patients. Rapid H. pylori cassette test results of the 224 patients were negative for 158 (70.5%) patients and positive for 66 (29.5%) patients, whereas micro ELISA IgA test results were negative for 110 (49.1%) patients and positive for 114 (50.9%) patients. Micro ELISA IgG test results were negative for 85 (37.9%) patients and positive for 139 (62.1%) patients. Conclusions: Invasive diagnostic methods for H. pylori infection may sometimes be inconvenient, and therefore the diagnosis may have to rely on non-invasive tests. Bases on the study results, we believe micro ELISA test results are more reliable with regard to avoidance of missed diagnosis. Keywords: Helicobacter pylori, Rapid casette test, ELISA, Gastric ulcer


2019 ◽  
Author(s):  
M.Victoria Esteban-Mendoza ◽  
Víctor Arcila-Quiceno ◽  
J. Albarracín-Navas ◽  
Isabel Hernández ◽  
C. Flechas Alarcón ◽  
...  

Abstract Background: Cardiopulmonary dirofilariosis caused by Dirofilaria immitis is a zoonotic, vector-borne infection, whose main hosts are both domestic and wild canids and which can be transmitted to humans. It mainly occurs in tropical and subtropical climates, and temperature and humidity are the main factors that favor the presence and proliferation of Culicidae mosquitoes. There have been few reports of this disease in dogs and humans in Colombia, a region with favorable climatic conditions for the development of this disease. Therefore, this research aimed to study its current prevalence in dogs and the risk of human exposure to the disease in Bucaramanga, one of the most populated areas in Colombia located at the center of the country. Furthermore, its demographic and environmental characteristics could be useful as a study model for other similar locations and neighboring countries. Method: Serum samples from 351 dogs and 506 humans from the Bucaramanga metropolitan area were analyzed. All dog samples were tested with a commercial immunochromatographic test kit to detect the presence of circulating antigens of D. immitis. Human samples were analyzed using a non-commercial ELISA test kit to detect IgG against the somatic antigens of adult D. immitis and Wolbachia. Positive results were further confirmed using a Western blot analysis. Results: Dirofilaria immitis prevalence was 10.82% (95% CI) in dogs and 5.12% (95% CI) in microfilaremic dogs. Seroprevalence in humans was 6.71% (95% CI) and was significantly higher in individuals aged 16–34 years and in women than in men. Conclusions: This study describes seropositivity to D. immitis for the first time in a Colombian human population located in the same area as that of dogs infected with D. immitis, which represents a potential threat to public health. In humans, age and sex can be considered risk factors for exposure to D. immitis.


2017 ◽  
Vol 14 (2) ◽  
pp. 215-220
Author(s):  
G. P. Yadav ◽  
S. Manadhar ◽  
A. K. Karna ◽  
K. P. Acharya ◽  
D. Maharjan ◽  
...  

Dairy industry is growing in Nepaland western dairy pocket area is the main milk producing area in Chitwan district. This study was carried out to determine the seroprevalence and risk factors associated with N. caninum in western dairy pocket area in Chitwan district of Nepal. A cross-sectional study was conducted from April 2014 to July 2014 among the small holder dairy farms, and out of 2188 animals from different sampling villages, 186 (8.5%) animals were randomly selected. Individual blood samples (5-10 ml) were collected and screened for N. caninum antibodies by ELISA test kit. The study showed that apparent overall prevalence of N. caninum to be 4.84% and true prevalence, 4.07%. The prevalence varied from a low of 0% to a high of 13.16% in various VDCs. The prevalence rates among Holstein-Friesian cross and Jersey cross Cattle were 6.94% and 3.51% respectively. The seroprevalence of N. caninum was found to be 16.13% and 2.5% in animals with and without history of abortion respectively, showed statistically significant different (p value <0.05). Similarly, prevalence were 13.64% and 3.66% in animals with and without presence of dog respectively and 8.5%, 3.39% and 0% in the  animals of age group of 3-5 years, 1-3 years and above 5 years respectively, showed statistically non-significant association (p value <0.05). This study shows that N. caninum is associated with abortion in dairy cattle of Chitwan region. The study may contribute the base line data of N. caninum in Nepal for future preventive strategy for stake-holders and government.


2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Daniela Proverbio ◽  
Eva Spada ◽  
Luciana Baggiani ◽  
Giada Bagnagatti De Giorgi ◽  
Roberta Perego

This study compares a rapid Immunospecific Kalazar Canine Rapid Spot IF with the gold standard test (indirect fluorescent antibody test (IFAT)) for detection ofLeishmania infantumspecific IgG serum antibodies in naturally exposed dogs. Serum samples were obtained from 89 healthy dogs and dogs affected by canine leishmaniosis (CanL). IgG-IFAT titers ≥80 were considered positive. Anti-L. infantumIgG antibodies were found in 54 samples with titers ranging from 1 : 80 to 1 : 5120. The performance of the rapid Immunospecific Kalazar was evaluated using a ROC curve. The area under the ROC curve of 0.957 was significantly different from 0.5 (), and therefore it can be concluded that the rapid Immunospecific Kalazar has the ability to distinguish canine sera with and withoutL. infantumIgG. The best performance of the test was at a cutoff >0 (sensitivity 92.6%, specificity 97%). The test can be used for disease screening if the cutoff is >0 (highest sensitivity, 92.6%) and is recommended as confirmatory test for the presence ofL. infantumIgG antibodies if the cutoff is set >2 (highest specificity, 100%).


2016 ◽  
Vol 14 (1) ◽  
pp. 65-68 ◽  
Author(s):  
A. Chakrabartty ◽  
P. K. Bhattacharjee ◽  
R. R. Sarker ◽  
A. K. M. A. Rahman ◽  
K. Henning ◽  
...  

The objectives of this study were to determine the prevalence of Coxiella burnetii infection in domestic ruminants and to detect Coxiella burnetii DNA from ticks and serum samples. A total of 24 ticks, 91 goats and 81 cattle serum samples with the history of abortion and reproductive disorders were collected from the different areas in Bangladesh. The serum samples were tested by CHEKIT Q-Fever Antibody ELISA Test Kit and Coxiella burnetii DNA was detected by multiplex quantitative real- time PCR. The overall prevalence was 7.6% and 6.1% in goats and cattle, respectively. However, none of seropositive samples and tick samples was positive in quantitative real-time PCR.


1994 ◽  
Vol 13 (10) ◽  
pp. 683-688 ◽  
Author(s):  
Françoise Audebert ◽  
Marc Sorkine ◽  
Annie Robbe-Vincent ◽  
Cassian Bon

1 A second inquiry was conducted in France to collect more accurate epidemiological, clinical and biological data from patients hospitalized after a viper bite, as well as treatment that they received. Fifty-seven well documented cases were classified in four grades of increasing severity defined according to the clinical signs of envenomation. 2 Local and systemic signs of envenomation appeared during the first 3 h, but the severity of the envenomation was observed to increase during the 12-24 h following bites in 50% of moderate and severe cases. One fatal case was reported. Biological analysis revealed an hyperleukocytosis in patients with moderate and severe envenomations. 3 The average length of hospitalization was of 1.7 ± 1.3 days for patients without signs of envenomation (grade 0) or presenting a minimal envenomation (grade 1), and statistically longer, 6.2 ± 2.9 days, for patients presenting moderate (grade 2) or severe envenomation (grade 3). 4 Levels of venom antigens in serum samples regularly collected during hospitalization were determined by a sandwich ELISA test. The serum venom levels determined during the first 4 h following the bite correlated with the severity of the envenomation when the symptoms were determined at their worst, usually 12-24 h later. In fact, concentrations higher than 20 ng ml-1 predict a moderate or severe clinical evolution. 5 The pharmacokinetics of venom antigens was also investigated during human envenomations. Venom antigens could be detected in the serum of patients as soon as half an hour after the bite, then decreased exponentially during the following hours with an apparent half-life of 8 h, similar in grade 2 and grade 3 patients, indicating that the kinetics of venom antigens in blood is independent of the severity of the envenomation.


2016 ◽  
Vol 2 (7) ◽  
pp. 70 ◽  
Author(s):  
Hams Hussain Hashim Al-Fattli

Objectives This study was conducted to disclose the specific antibodies against M. bovis of bovine tuberculosis (bTB) in blood and milk serum samples, with detection of the most prevalent clinical signs in positive cows.Methods In some rural areas of two Iraqi provinces (Wasit and Dhi-Qar), 119 lactating cows were submitted to the clinical examination with obtaining of blood and milk to tested by using the IDEXX ELISA test.Results The overall seroprevalence in blood and milk was (20.16%) and (15.12%), respectively. In Wasit, the prevalence was (22.85%) and (15.71%), while in Dhi-Qar, the prevalence was (16.32%) and (14.28%) in blood and milk, respectively. As well as, marked significant differences in seroprevalence were observed between and within the two study’s provinces and samples. According to clinical examination, a significant rising (P > 0.05) was revealed in respiratory disorders, decreasing in milk production, emaciation, rough hair coat and repetitive reproductive problems, whilst a significant decreasing (P < 0.05) in persistent feces abnormalities, mastitis, lymph nodes enlargement and loss of appetite.Conclusion The study demonstrated, for the first time in Iraq, the efficient of IDEXX ELISA, as a screening test in the detection of bTB in lactating cows by using blood and milk serum samples, and the competence of milk, as sample, in exhibition of infection. Also, the study exposed the high infection rate of bTB in cows of rural areas of Wasit and Dhi-Qar provinces.


Sign in / Sign up

Export Citation Format

Share Document