scholarly journals Development of a Multiplex PCR and Magnetic DNA Capture Assay for Detecting Six Species Pathogens of the Genera Anaplasma and Ehrlichia in Canine, Bovine, Caprine and Ovine Blood Samples from Grenada, West Indies

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 192
Author(s):  
Bhumika Sharma ◽  
Roman R. Ganta ◽  
Diana Stone ◽  
Andy Alhassan ◽  
Marta Lanza-Perea ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Clinical Signs ◽  
Small Ruminants ◽  
High Sensitivity ◽  
Molecular Evidence ◽  
Blood Samples ◽  
Persistent Problem ◽  
Magnetic Capture ◽  
Low Sensitivity ◽  
Species Specific

Infections with tick-borne pathogens belonging to Anaplasma/Ehrlichia in various vertebrate hosts are a persistent problem resulting in nonspecific clinical signs during early infection. Diagnosis of single and multi-infections with these pathogens, causing diseases in companion/agricultural animals and people, remains a challenge. Traditional methods of diagnosis, such as microscopy and serology, have low sensitivity and specificity. Polymerase chain reaction (PCR) assays are widely used to detect early-phase infections, since these have high sensitivity and specificity. We report the development and validation of an assay involving PCR followed by magnetic capture method using species-specific oligonucleotides to detect six Anaplasma/Ehrlichia species pathogens in canine, bovine, caprine, and ovine blood samples. Overall, the assay application to 455 samples detected 30.1% (137/455) positives for one or more out of six screened pathogens. Single-pathogen infections were observed in 94.9% (130/137) of the positive samples, while co-infections were detected in 5.1% (7/137). Anaplasma marginale infection in cattle had the highest detection rate (34.4%), followed by canines positive for Anaplasma platys (16.4%) and Ehrlichia canis (13.9%). The assay aided in documenting the first molecular evidence for A. marginale in cattle and small ruminants and Ehrlichia chaffeensis and Ehrlichia ewingii in dogs in the Caribbean island of Grenada.

scholarly journals Comparison of diagnostics of bacterial vaginosis according to clinical signs with results of laboratory investigations

2016 ◽  
Vol 65 (4) ◽  
pp. 76-82
Author(s):  
Elena V. Shipitsina ◽  
Tatyana A. Khusnutdinova ◽  
Olga S. Ryzhkova ◽  
Anna A. Krysanova ◽  
Olga V. Budilovskaya ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
Real Time ◽  
Sensitivity And Specificity ◽  
Real Time Pcr ◽  
Vaginal Discharge ◽  
Clinical Signs ◽  
High Sensitivity ◽  
Clinical Diagnostics ◽  
Laboratory Diagnostics ◽  
Molecular Method

Introduction. Bacterial vaginosis (BV) is associated with a number of reproductive health disorders, therefore timely and accurate diagnosis of this condition is exceedingly important. Objective.Comparison of effectiveness of clinical and laboratory diagnostics of BV in women with vaginal discharge. Material and methods. In total, 318 patients addressing gynecological clinics with complaints about vaginal discharge participated in the study. Clinical diagnostics of BV was performed in the clinics participating in patient enrollment in accordance with their clinical practice. For laboratory diagnostics, microscopy of Gram stained smears according to the Nugent method and quantitative real-time PCR were used. Sensitivity and specificity of clinical diagnostics of BV and the molecular method were evaluated using the Nugent method as reference standard. Results. With the Nugent method, BV was diagnosed in 27% of women, with real-time PCR — in 37% of women. Using clinical signs of BV, the condition was diagnosed in 91% women. Sensitivity and specificity of the real-time PCR were 97% and 87%, respectively. Sensitivity of clinical diagnostics was 100%, but specificity was only 17%. Conclusions. Diagnostics of BV based only on the presence of vaginal discharge leads to false positive results and requires laboratory confirmation. The molecular method has a high sensitivity and satisfactory specificity for BV diagnosis and can be used as an alternative to the Nugent method.

scholarly journals Role of endoscopy in suspicion of atrophic gastritis with and without intestinal metaplasia in comparison to histopathology

2021 ◽  
Vol 84 (1) ◽  
pp. 9-17
Author(s):  
H Ibrahim ◽  
A Shams El-Deen ◽  
ZA Kasemy ◽  
M Saad ◽  
AA Sakr
Keyword(s):  
Atrophic Gastritis ◽  
Intestinal Metaplasia ◽  
Sensitivity And Specificity ◽  
Chronic Gastritis ◽  
Gastrointestinal Symptoms ◽  
High Sensitivity ◽  
High Specificity ◽  
Poor Correlation ◽  
Gastric Lesions ◽  
Low Sensitivity

Background and study aims : Atrophic gastritis (AG) and intestinal metaplasia (IM) are established premalignant gastric lesions. Many studies documented a poor correlation between esophagogastroduodenoscopy (EGD) and histopathological (HP) findings of precancerous gastric lesions. The aim was to bridge the gap between endoscopy and HP in detection of chronic gastritis, AG and IM. Patients and methods : a prospective single-center study involved 150 patients with endoscopic criteria of gastric lesions with upper gastrointestinal symptoms referred for upper GI endoscopy met the endoscopic criteria and classified according to HP of biopsies from targeted gastric lesions into chronic gastritis (GI), AG(GII) or IM(GIII). We correlated the endoscopic criteria of the 3 groups with the HP results. Results : (73males & 75 females) with ages ranged17-75 years and mean± SD was 41.96 ± 15.95. GI, GII &GIII were [42 patients (28%),82 patients (54.7%) and 26 patients (17.3%)], respectively. Diffuse gastric mottling was more common in GI (74.3%, P<0.001), visible submucosal vessels, gastric atrophy predominated in GII (75.6, 82.3 & 73.1% (P 0.005,0.4 & <0.01)), respectively. Whitish raised lesions were more specific in GIII (85.7%) (P<0.001). The sensitivity and specificity of endoscopic suspicion of chronic gastritis were (86&88% in GI), (87&85% in GII) and (54% &100% in GIII) (p-0.001). The logistic regression model for risk factors was χ2= 25.74 and 49.32, p < 0.001. Conclusion : Conventional endoscopy has high sensitivity and specificity for suspicion of chronic gastritis and AG, but low sensitivity and very high specificity for IM. Targeted biopsies may be valuable with image enhanced techniques.

scholarly journals Fluorescent bead-based serological detection of Toxoplasma gondii infection in chickens

2020 ◽  
Author(s):  
Benedikt T. Fabian ◽  
Fatima Hedar ◽  
Martin Koethe ◽  
Berit Bangoura ◽  
Pavlo Maksimov ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Small Ruminants ◽  
High Sensitivity ◽  
Negative Control ◽  
Immunofluorescence Assay ◽  
Farm Animals ◽  
Serological Tests ◽  
Modified Agglutination Test ◽  
Magnetic Capture ◽  
Luminex Technology

Abstract Background: Free-ranging chickens are often infected with Toxoplasma gondii. Their infection indicates environmental contamination with T. gondii. The detection of infected birds relies primarily on serological assays. Methods: Here, we established a bead-based multiplex assay (BBMA) using the Luminex technology for the specific and sensitive detection of T. gondii infections in chickens. Recombinant biotinylated T. gondii surface antigen 1 (TgSAG1bio) bound to streptavidin-conjugated magnetic Luminex beads served as antigen. Serum antibodies were detected by a fluorophore-coupled secondary antibody. Beads of differing color codes were conjugated with anti-chicken IgY or chicken serum albumin and served for each sample as an internal positive or negative control, respectively. The assay was validated with sera from experimentally and naturally infected chickens. The results were compared to those from reference methods, including other serological tests and bioassay in mice.Results: In experimentally infected chickens, the vast majority (98.5%, n=65/66) of inoculated birds tested seropositive in the BBMA. This included all chickens positive by magnetic-capture PCR (100%, n=45/45). Most, but not all inoculated and TgSAG1bio-BBMA-positive chickens were also positive in two previously established TgSAG1-ELISAs (TgSAG1-ELISASL, n=61/65; or TgSAG1-ELISASH, n=60/65), or positive in an immunofluorescence assay (IFAT, n=64/65)) and in a modified agglutination test (MAT, n=61/65). All non-inoculated control animals (n=28/28, 100%) tested negative. In naturally exposed chickens, the TgSAG1bio-BBMA showed a high sensitivity (98.5%; 95% Confidence Interval: 90.7-99.9%) and specificity (100%; 85.0-100%) relative to a reference standard established using ELISA, IFAT and MAT. Almost all naturally exposed chickens that were positive in bioassay or by PCR tested positive in the TgSAG1bio-BBMA (93.5%; 77.1-98.9%), while all bioassay- or PCR-negative chickens remained negative (100%; 85.0-100%).Conclusions: The TgSAG1bio-BBMA represents a suitable method for the detection of T. gondii infections in chickens with high sensitivity and specificity, which is comparable or even superior to other tests. Since assays based on this methodology allow for the simultaneous analysis of a single biological sample with respect to multiple analytes, the described assay may represent one component in future multiplex assays for broad serological monitoring of poultry and other farm animals, including pigs or small ruminants, for various pathogens.

Investigations on First Confirmed Outbreak of Ovine Theileriosis (Theileria luwenshuni) from Maharashtra State, India

2020 ◽  
Author(s):  
V.S. Dhaygude ◽  
K. Kundu ◽  
B.P. Kamdi ◽  
U.R. Bagal ◽  
S.B. Bhosale ◽  
...  
Keyword(s):  
18S Rrna ◽  
Clinical Signs ◽  
Specific Treatment ◽  
Small Ruminants ◽  
18S Rrna Gene ◽  
Pcr Detection ◽  
Nucleotide Sequencing ◽  
Rrna Gene ◽  
Blood Samples ◽  
Theileria Luwenshuni

Background: Clinical theileriosis of small ruminants is tick-borne disease caused by Theileria lestoquardi, Theileria uilenbergi and Theileria luwenshuni. Theileria annulata, the causative agent of bovine tropical theileriosis in cattle, can also infect sheep but does not cause any significant illness. It is one of the economically important diseases. There are no reports of ovine clinical theileriosis from Maharashtra state and there is paucity of information on its epidemiology. This paper reports first confirmed outbreak of ovine theileriosis based on clinical signs, microscopic examination, PCR and sequencing in the Maharashtra State of India. Methods: Whole blood samples from 22 ailing sheep were collected and subjected to hematological examination. Blood smears stained with Leishman’s stain were examined under 100X objective of the microscope. The blood samples from sheep found positive by microscopic method were subjected to PCR detection of 18S rRNA gene of hemoprotozoa and then for nucleotide sequencing and sequence analysis.Conclusion: Samples from 14 out of 22 sheep were found positive for piroplasms of Theileria spp by light microscopy. All positive samples were further confirmed by PCR detection of 18S rRNA gene of hemoprotozoa. PCR amplification yielded expected product of 1750 bp for all samples. BLAST and phylogenetic analysis of one sample revealed high sequence homology with T. luwenshuni reported from India and other countries. Characteristic clinical signs like fever, progressive anaemia, laboured breathing, lymphadenopathy, debility and non-responsiveness to antibiotic therapy were recorded. The animals responded to specific treatment against theileriosis. It is the first ever confirmed report of ovine theileriosis in Maharashtra state of India and hence reported.

scholarly journals Sensitivity and specificity of loss of heterozygosity analysis for the classification of rare germline variants in BRCA1/2: results of the observational AGO-TR1 study (NCT02222883)

2020 ◽  
pp. jmedgenet-2020-107353
Author(s):  
Jan Hauke ◽  
Philipp Harter ◽  
Corinna Ernst ◽  
Alexander Burges ◽  
Sandra Schmidt ◽  
...  
Keyword(s):  
Loss Of Heterozygosity ◽  
Sensitivity And Specificity ◽  
False Negative ◽  
High Sensitivity ◽  
High Specificity ◽  
Germline Variants ◽  
Variants Of Unknown Significance ◽  
Link Type ◽  
Registration Number ◽  
Low Sensitivity

Variant-specific loss of heterozygosity (LOH) analyses may be useful to classify BRCA1/2 germline variants of unknown significance (VUS). The sensitivity and specificity of this approach, however, remains unknown. We performed comparative next-generation sequencing analyses of the BRCA1/2 genes using blood-derived and tumour-derived DNA of 488 patients with ovarian cancer enrolled in the observational AGO-TR1 trial (NCT02222883). Overall, 94 pathogenic, 90 benign and 24 VUS were identified in the germline. A significantly increased variant fraction (VF) of a germline variant in the tumour indicates loss of the wild-type allele; a decreased VF indicates loss of the variant allele. We demonstrate that significantly increased VFs predict pathogenicity with high sensitivity (0.84, 95% CI 0.77 to 0.91), poor specificity (0.63, 95% CI 0.53 to 0.73) and poor positive predictive value (PPV; 0.71, 95% CI 0.62 to 0.79). Significantly decreased VFs predict benignity with low sensitivity (0.26, 95% CI 0.17 to 0.35), high specificity (1.0, 95% CI 0.96 to 1.00) and PPV (1.0, 95% CI 0.85 to 1.00). Variant classification based on significantly increased VFs results in an unacceptable proportion of false-positive results. A significantly decreased VF in the tumour may be exploited as a reliable predictor for benignity, with no false-negative result observed. When applying the latter approach, VUS identified in four patients can now be considered benign. Trial registration numberNCT02222883.

scholarly journals Diagnosis and Pathogenesis of Nairobi Sheep Disease Orthonairovirus Infections in Sheep and Cattle

Viruses ◽  
2021 ◽  
Vol 13 (7) ◽  
pp. 1250
Author(s):  
Julia Hartlaub ◽  
Benjamin Gutjahr ◽  
Christine Fast ◽  
Ali Mirazimi ◽  
Markus Keller ◽  
...  
Keyword(s):  
Climate Change ◽  
Clinical Signs ◽  
Small Ruminants ◽  
Virus Shedding ◽  
Diagnostic Assays ◽  
Virus Dose ◽  
Severe Gastroenteritis ◽  
Species Specific ◽  
Tick Vectors ◽  
Different Doses

Nairobi sheep disease orthonairovirus (NSDV) is a zoonotic tick-borne arbovirus, which causes severe gastroenteritis in small ruminants. To date, the virus is prevalent in East Africa and Asia. However, due to climate change, including the spread of transmitting tick vectors and increased animal movements, it is likely that the distribution range of NSDV is enlarging. In this project, sheep and cattle (hitherto classified as resistant to NSDV) were experimentally infected with NSDV for a comparative study of the species-specific pathogenesis. For this purpose, several new diagnostic assays (RT-qPCR, ELISA, iIFA, mVNT, PRNT) were developed, which will also be useful for future epidemiological investigations. All challenged sheep (three different doses groups) developed characteristic clinical signs, transient viremia and virus shedding—almost independent on the applied virus dose. Half of the sheep had to be euthanized due to severe clinical signs, including hemorrhagic diarrhea. In contrast, the course of infection in cattle was only subclinical. However, all ruminants showed seroconversion—implying that, indeed, both species are susceptible for NSDV. Hence, not only sheep but also cattle sera can be included in serological monitoring programs for the surveillance of NSDV occurrence and spread in the future.

scholarly journals Detection And Molecular Characterization of Theileria Ovis in Sheep And Goats with Clinical The Ileriosis in Kurdistan, Iraq

2020 ◽  
Vol 23 (2) ◽  
pp. 69-78
Author(s):  
Zuber Ismael Hassen ◽  
Azad Abdullah Meerkhan
Keyword(s):  
Disease Transmission ◽  
Clinical Signs ◽  
Kurdistan Region ◽  
Single Test ◽  
Molecular Method ◽  
Blood Samples ◽  
Sheep And Goats ◽  
Specific Pcr ◽  
Species Specific

This study was carried out to detect Theileria infection in sheep and goats in Kurdistan region, Iraq from June 2019 to April 2020. Molecular method was used to identify Theileria species. Sixty- seven blood samples were taken from 45 sheep and 22 goats based on clinical signs of theileriosis during tick activating season. The 67 samples were PCR edm and as a result, 20 species-specific PCR were positives (26.67% (12/20) were Theileria ovis in sheep and 36.36% (8/20) were from goats). The results of the gene analysis in the current study were registered in NCBI under four accession numbers (MN889986, MN889987, MN889988 and MN889989), which shows that sheep and goats can be infected with Theileria ovis. This is the first report of Theileria species in goats with clinical theileriosis in Kurdistan, so the gene flow and disease transmission between sheep and goats is most expected. PCR is a useful diagnostic tool to detect ovine theileriosis with a single test and suggested that T. ovis is the dominant piroplasmid agent in Erbil. In addition, it revealed that sheep is very susceptible to theileriosis than goats

scholarly journals Sensitivity and Specificity of Sheard and Percival’s Criteria for the Diagnosis of Young People with Near-heterophoria

2021 ◽  
Vol 9 (B) ◽  
pp. 1128-1134
Author(s):  
Saif Hassan Alrasheed ◽  
Amel Mohamed Yousif ◽  
Majid A. Moafa ◽  
Abd Elaziz Mohamed Elmadina ◽  
Mohammad Alobaid
Keyword(s):  
Young People ◽  
Negative Predictive Value ◽  
Sensitivity And Specificity ◽  
Binocular Vision ◽  
Predictive Value ◽  
High Sensitivity ◽  
The Other ◽  
Cross Sectional ◽  
Predictive Values ◽  
Low Sensitivity

BACKGROUND: Sheard and Percival assumed that symptoms from latent strabismus can be avoided if the relevant fusional vergence is adequate to support the heterophoria. AIM: The aim of the study was to determine the sensitivity and specificity of Sheard’s and Percival’s criterion for the diagnosis of heterophoria. METHODS: A cross-sectional hospital-based study was performed at Al-Neelain Eye Hospital Khartoum, Sudan from February to October 2019. Heterophoria was measured using Maddox Wing and fusional vergence using a prism bar. Thereafter, Sheard’s and Percival’s criteria were used for the diagnosis of heterophoria. RESULTS: A total of 230 participants (age = 15–30 years; mean age = 19.34 ± 3.325 years) were recruited for this study. The Sheard’s criteria showed a high sensitivity of 87.2% and a low specificity of 8.0% for the diagnosing of exophoria, with positive and negative predictive values of 65.5% and 26%, respectively. The criteria showed a relatively low sensitivity of 77.8% and a specificity of 9.0% in the diagnosis of esophoria, with a positive and negative predictive values of 56% and 20%, respectively. Percival criteria showed high sensitivity 84.2% and low specificity 9.1% in diagnosing esophoria, with a positive and negative predictive value of 61.5% and 25%, respectively. On the other hand, the criteria showed low sensitivity 67.4% and specificity 13.8% in diagnosing exophoria, with positive and negative predictive value 61.9% and 17%, respectively. CONCLUSION: Sheard’s and Percival’s criteria are useful in diagnosing binocular vision problems. Sheard’s criteria are accurate in diagnosing near exophoria and Percival’s criteria are more accurate in diagnosing near esophoria. Therefore, these criteria provide good clues and predictions for the diagnosis of binocular vision problems.

scholarly journals A Rapid In-Clinic Test Detects Acute Leptospirosis in Dogs with High Sensitivity and Specificity

2016 ◽  
Vol 2016 ◽  
pp. 1-3 ◽  
Author(s):  
Angeli Kodjo ◽  
Christophe Calleja ◽  
Michael Loenser ◽  
Dan Lin ◽  
Joshua Lizer
Keyword(s):  
Sensitivity And Specificity ◽  
Point Of Care ◽  
Clinical Signs ◽  
High Sensitivity ◽  
Diagnostic Sensitivity ◽  
Immunochromatographic Test ◽  
Serum Samples ◽  
Weight Group ◽  
Agglutination Assay ◽  
Diagnostic Sensitivity And Specificity

A rapid IgM-detection immunochromatographic test (WITNESS® Lepto, Zoetis) has recently become available to identify acute canine leptospirosis at the point of care. Diagnostic sensitivity and specificity of the test were evaluated by comparison with the microscopic agglutination assay (MAT), using a positive cut-off titer of ≥800. Banked serum samples from dogs exhibiting clinical signs and suspected leptospirosis were selected to form three groups based on MAT titer: (1) positive (n=50); (2) borderline (n=35); and (3) negative (n=50). Using an analysis to weight group sizes to reflect French prevalence, the sensitivity and specificity were 98% and 93.5% (88.2% unweighted), respectively. This test rapidly identifies cases of acute canine leptospirosis with high levels of sensitivity and specificity with no interference from previous vaccination.

scholarly journals Isolation and Cultivation of a New Isolate of BTV-25 and Presumptive Evidence for a Potential Persistent Infection in Healthy Goats

Viruses ◽  
2020 ◽  
Vol 12 (9) ◽  
pp. 983
Author(s):  
Christina Ries ◽  
Ursula Domes ◽  
Britta Janowetz ◽  
Jens Böttcher ◽  
Katinka Burkhardt ◽  
...  
Keyword(s):  
Clinical Signs ◽  
Small Ruminants ◽  
Full Genome ◽  
Serum Samples ◽  
Blood Samples ◽  
Experimental Inoculation ◽  
Serological Surveillance ◽  
First Time ◽  
Goat Flock ◽  
Edta Blood

Recently, several so-called “atypical” Bluetongue virus (BTV) serotypes were discovered, including BTV-25 (Toggenburg virus), in Switzerland. Most “atypical” BTV were identified in small ruminants without clinical signs. In 2018, two goats from a holding in Germany tested positive for BTV-25 genome by RT-qPCR prior to export. After experimental inoculation of the two goats with the BTV-25 positive field blood samples for generation of reference materials, viremia could be observed in one animal. For the first time, the BTV-25-related virus was isolated in cell culture from EDTA-blood and the full genome of isolate “BTV-25-GER2018” could be generated. BTV-25-GER2018 was only incompletely neutralized by ELISA-positive sera. We could monitor the BTV-25 occurrence in the respective affected goat flock of approximately 120 goats over several years. EDTA blood samples were screened with RT-qPCR using a newly developed BTV-25 specific assay. For serological surveillance, serum samples were screened using a commercial cELISA. BTV-25-GER2018 was detected over 4.5 years in the goat flock with intermittent PCR-positivity in some animals, and with or without concomitantly detected antibodies since 2015. We could demonstrate the viral persistence of BTV-25-GER2018 in goats for up to 4.5 years, and the first BTV-25 isolate is now available for further characterization.

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