scholarly journals Further evaluation of recombinant Tsol-p27 by enzyme-linked immunoelectrotransfer blot for the serodiagnosis of cysticercosis in pigs from Mozambique

2019 ◽  
Vol 12 (1) ◽  
Author(s):  
Noemia Nhancupe ◽  
Emilia V. Noormahomed ◽  
Sonia Afonso ◽  
Staffan Svard ◽  
Johan Lindh
Keyword(s):  
Sensitivity And Specificity ◽  
Predictive Value ◽  
Negative Impact ◽  
Low Cost ◽  
High Sensitivity ◽  
Epileptic Seizures ◽  
Enzyme Linked Immunosorbent Assay ◽  
Meat Industry ◽  
Porcine Cysticercosis ◽  
Human Sera

Abstract Background Porcine cysticercosis has a negative impact on human health and the meat industry, as it makes infected meat unaproprieted for consuption and it is the main etiology of epileptic seizures in developing countries. There are multiple serological assays that use crude antigens with high sensitivity and specificity for the diagnosis of both porcine and human cysticercosis. Nonetheless, antigen preparation is time-consuming, needs a well-equipped laboratory and trained personnel and places those manipulating the meat at great risk for infection. New serodiagnostic approaches to the diagnosis of porcine and human cysticercosis have been directed towards the development of recombinant deoxyribonucleic acid technology for the generation of synthetic proteins that can serve as simplified, low-cost and harmless substitutes for native antigens. The aim of the present study was to further evaluate the recombinant Tsol-p27 protein as a target molecule in immunoassays for the serodiagnosis of porcine cysticercosis. From these data, we hoped to develop recommendations regarding its use in the serodiagnosis of porcine cysticercosis. Results We studied a panel of 83 naturally infected pig sera from Angónia District, Mozambique, an endemic area for porcine and human cysticercosis. These sera were previously tested by antigen enzyme-linked immunosorbent assay (Ag-ELISA) to detect antigens of T. solium. The serum panel was processed by enzyme-linked immunoelectrotransfer blot (EITB) assay against the recombinant Tsol-p27 protein and the Ag-ELISA assay results were used to compare and evaluate the performance of Tsol-p27 for the diagnosis of cysticercosis. Out of 83 sera, 24 (29.0%) were positive for Tsol-p27 and 59 (71%) were negative in the same assay. From the 37 sera that tested positive to Ag-ELISA, 11 (13.3%) were positive to Tsol-p27, while from 46 sera that tested negative to Ag-ELISA, 33 (39.7%) also tested negative to Tsol-p27. The sensitivity and specificity of Tsol-p27 was 29.7% and 71.7%, respectively, while the positive predictive value and negative predictive value were 45.8% and 55.9%, respectively, as calculated using Medcalc® version 15.0 software (MedCalc Software, Ostend, Belgium). Conclusion While Tsol-p27 recombinant protein might be suitable for testing human sera, its performance in pigs is not acceptable, so other recombinant proteins should be evaluated alone or multiplexed.

scholarly journals Performance of Tsol-p27 antigen for the serological diagnosis of cysticercosis in Mozambique

2015 ◽  
Vol 90 (5) ◽  
pp. 630-633 ◽  
Author(s):  
N. Nhancupe ◽  
E.V. Noormahomed ◽  
S. Afonso ◽  
K.I. Falk ◽  
J. Lindh
Keyword(s):  
Western Blot ◽  
Sensitivity And Specificity ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Low Cost ◽  
High Sensitivity ◽  
Diagnostic Methods ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Neuroimaging Techniques

AbstractThe diagnosis of neurocysticercosis (NCC) requires expensive neuroimaging techniques that are seldom affordable for people in endemic countries. Accordingly, there is a need for new low-cost diagnostic methods that offer high sensitivity and specificity. In this study, we evaluated Western blot analysis of the previously described recombinant antigen Tsol-p27 in relation to a commercial or in-house enzyme-linked immunosorbent assay (ELISA) for NCC, and compared the results with those provided by a commercial enzyme-linked immunoelectrotransfer blot (EITB) assay, which was regarded as the reference standard method. The analysed serum samples were obtained from 165 people, 18 of whom were confirmed to be NCC positive by EITB. Comparing our Western blot analysis of Tsol-p27 with a previous evaluation performed in Central America showed similar specificity (96.69% versus 97.8%) and sensitivity (85.71% versus 86.7%). The present results indicate that the recombinant Tsol-p27 antigen provides good sensitivity and specificity, and might be preferable as a diagnostic antigen in poorly equipped laboratories in endemic countries.

scholarly journals A new ELISA and western blot technique based on recombinant TES antigen and/or larval antigen for the detection of toxocariasis in humans

Parasitology ◽  
2020 ◽  
pp. 1-8
Author(s):  
Marie-Kristin Raulf ◽  
Daniela Jordan ◽  
Herbert Auer ◽  
Jens M. Warnecke ◽  
Bernd Lepenies ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
High Reliability ◽  
High Sensitivity ◽  
Cross Reactivity ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Tests ◽  
Western Blots ◽  
Blot Technique ◽  
Human Sera

Abstract Serological antibody detection by enzyme-linked immunosorbent assay (ELISA)- and immunoblot-based methods constitutes the best indicator of human Toxocara infection. Nevertheless, the availability of serological tests, particularly western blots (WB), evaluated for sensitivity and specificity is limited. Therefore, an Anti-Toxocara-ELISA immunoglobulin g (IgG) prototype (Proto-ELISA) and an Anti-Toxocara-Westernblot (IgG) prototype (Proto-WB) were evaluated by testing 541 human sera pre-determined for Toxocara infection by an established in-house Anti-Toxocara-ELISA (IH-ELISA). To evaluate sensitivity and specificity of the newly developed ELISA and WB prototypes, results were compared to IH-ELISA and a commercial WB (Com-WB). Compared to the IH-ELISA, a sensitivity of 93.1% (229/246) and a specificity of 94.6% (279/295) of the Proto-ELISA with a Cohen's κ of 0.88 were obtained. The sensitivity of the Proto-WB was 76.7% (240/313) and specificity was 99.6% (227/228) with a Cohen's κ of 0.73 compared to those of Com-WB. A comparison to the IH-ELISA revealed 91.5% (225/246) sensitivity and 94.6% (279/295) specificity of the Proto-WB with a Cohen's κ of 0.86. Cross-reactivity was observed for some samples positive for Ascaris and Trichinella spp. in the Proto-ELISA, Proto-WB and Com-WB. Overall, the evaluated ELISA and WB prototypes showed high sensitivity and specificity, indicating high reliability of these newly developed tests.

scholarly journals Development and standardization of the Dot Blot test for serological diagnosis of bovine leptospirosis

2021 ◽  
Vol 10 (6) ◽  
pp. e22710615091
Author(s):  
Higor Oliveira Silva ◽  
Mariana Assunção de Souza ◽  
Tatiane Cristina Fernandes Tavares ◽  
Pollyanna Mafra Soares ◽  
Anna Monteiro Correia Lima
Keyword(s):  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Predictive Value ◽  
Bovine Serum ◽  
Low Cost ◽  
High Sensitivity ◽  
Dot Blot ◽  
Blot Technique ◽  
Bovine Leptospirosis

Leptospirosis a public health problem and an endemic zoonosis in Brazil, is diagnosed by serological methods. Therefore, low-cost and easy to execute methodologies with good/high sensitivity, such as Dot Blot, are an important diagnostic tool. The aim of this study was to standardize and validation the dot-blot technique for the serological diagnosis of bovine leptospirosis. Several concentrations of antigens applied to nitrocellulose membranes, and different dilutions of conjugated bovine serum were evaluated to develop and standardize the test. The best distinction/contrast between positive and negative samples was observed for 1μL antigen (0.11μg/μL outer membrane protein of the Hardjo serovar (OMPH) and 0.08 μg/μL outer membrane protein of the Wolffi serovar (OMPW)), 1:500 and 1:10000 bovine serum dilution and conjugate, respectively. The Dot Blot test efficiency was 71.87% and kappa index, 0.46 (p<0.0001). The other parameters measured were: sensitivity 91.89%; specificity 59.32%; positive predictive value 58.62%; and, negative predictive value 59.32%. In addition to high sensitivity, other advantages of the Dot Blot technique have been identified, such as practicality, low cost since it does not require sophisticated devices and the fact that the Hardjo and Wolffi OMP also reacted with serovars from other pathogenic serogroups. The results provided positive expectations for the use of Dot Blot as support in the diagnosis of bovine leptospirosis, especially if used as a screening test, stimulating further research for the future development of kits for diagnostic purposes.

Standardization and characterization of antigens for the diagnosis of aspergillosis

2012 ◽  
Vol 58 (4) ◽  
pp. 455-462 ◽  
Author(s):  
Cheila Denise Ottonelli Stopiglia ◽  
Alicia Arechavala ◽  
Mariana Carissimi ◽  
Julia Medeiros Sorrentino ◽  
Valério Rodrigues Aquino ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Aspergillus Niger ◽  
Sensitivity And Specificity ◽  
Predictive Value ◽  
High Sensitivity ◽  
Antigenic Activity ◽  
Sds Page ◽  
Mascot Search Engine ◽  
Commercial Kit

The aim of this study was to develop and characterize antigens for the diagnosis of aspergillosis. Nine strains of Aspergillus species Aspergillus fumigatus , Aspergillus flavus , and Aspergillus niger were grown in Sabouraud and Smith broth to produce exoantigens. The antigens were tested by immunodiffusion against sera from patients with aspergillosis and other systemic mycoses. The protein fraction of the antigens was detected by SDS–PAGE; Western blot and representative bands were assessed by mass spectrometry coupled to a nano Acquity UltraPerformance LC and analyzed by the Mascot search engine. Concurrently, all sera were tested with Platelia Aspergillus EIA. The most reactive antigens to sera from patients infected by A. fumigatus were produced by A. fumigatus MG2 Sabouraud and pooled A. fumigatus Sabouraud samples, both with a sensitivity of 93% and specificity of 100% and 97%, respectively. Aspergillus niger and A. flavus antigens were reactive against A. niger and A. flavus sera, each one with a sensitivity and specificity of 100%. Two proteins, probably responsible for antigenic activity, β-glucosidase in A. fumigatus and α-amylase in A. niger were attained. The commercial kit had a specificity of 22%, sensitivity of 100%, positive predictive value of 48%, and negative predictive value of 100%. The antigens produced showed high sensitivity and specificity and can be exploited for diagnostics of aspergilloma.

scholarly journals The Clinical Utility of the Heparin Neutralization Assay in the Diagnosis of Heparin-Induced Thrombocytopenia

2017 ◽  
Vol 24 (5) ◽  
pp. 749-754 ◽  
Author(s):  
Gang Zheng ◽  
Michael B. Streiff ◽  
Clifford M. Takemoto ◽  
Jennifer Bynum ◽  
Elise Gelwan ◽  
...  
Keyword(s):  
Diagnostic Accuracy ◽  
Predictive Value ◽  
Clinical Utility ◽  
Predictive Accuracy ◽  
High Sensitivity ◽  
Neutralization Assay ◽  
Serotonin Release ◽  
Enzyme Linked Immunosorbent Assay ◽  
Heparin Induced Thrombocytopenia ◽  
Heparin Neutralization

Heparin-induced thrombocytopenia (HIT) remains diagnostically challenging. Immunoassays including PF4/heparin enzyme-linked immunosorbent assay (ELISA) have high sensitivity but low specificity. Whether the heparin neutralization assay (HNA) improves the diagnostic accuracy of the PF4/heparin ELISA for HIT is uncertain. In this study, to assess its clinical utility and evaluate whether it improves the diagnostic accuracy for HIT, we implemented HNA in conjunction with PF4/heparin ELISA over a 1-year period. A total of 1194 patient samples were submitted to the laboratory for testing from December 2015 to November 2016. Heparin neutralization assay alone is a poor predictor for HIT, but it has high negative predictive value (NPV): Cases with %inhibition <70% are always negative for serotonin release assay. It improves the diagnostic positive predictive value (PPV) of ELISA without compromising sensitivity: ELISA optical density (OD) ≥1.4 alone has a sensitivity of 88% (14/16) and a PPV of 61% (14/23); with HNA %inhibition ≥70%, the sensitivity remains 88% (14/16) and PPV is 82% (14/17). 4Ts score correlates with ELISA OD and predicts HIT; the predictive accuracy of 4Ts score is further improved by HNA. Interestingly, HNA %inhibition of <70% correlates with low 4Ts scores. Based on its high NPV, HNA has the potential to facilitate more timely and accurate HIT diagnosis.

scholarly journals Lung Ultrasound in the Screening of Pulmonary Interstitial Involvement Secondary to Systemic Connective Tissue Disease: A Prospective Pilot Study Involving 180 Patients

2021 ◽  
Vol 10 (18) ◽  
pp. 4114
Author(s):  
Natalia Buda ◽  
Anna Wojteczek ◽  
Anna Masiak ◽  
Maciej Piskunowicz ◽  
Wojciech Batko ◽  
...  
Keyword(s):  
Connective Tissue ◽  
Connective Tissue Disease ◽  
Sensitivity And Specificity ◽  
Predictive Value ◽  
Lung Ultrasound ◽  
Imaging Modality ◽  
Connective Tissue Diseases ◽  
High Sensitivity ◽  
Study Group ◽  
A Prospective Study

Objectives: The aim of the study was the assessment of lung ultrasound (LUS) as a screening of pulmonary interstitial involvement secondary to systemic connective tissue diseases. Methods: A prospective study was conducted on the study group comprising 180 patients diagnosed with different systemic connective tissue diseases. Each patient underwent lung ultrasound (LUS), high-resolution chest computed tomography (HRCT), and echocardiography (ECHO). Each imaging examination was blinded and performed by an independent operator. LUS was conducted with convex and linear transducers. Results: The sensitivity and specificity of LUS as compared to HRCT in detecting pulmonary interstitial involvement in the study group were 99.3% and 96.4%, respectively; positive predictive value (PPV) 0.7, negative predictive value (NPV) 3.6. Abnormalities indicating interstitial lung disease (ILD) with fibrosis were most frequently localized bilaterally in the lower fields of the lungs, assessed in the dorsal view. Conclusions: LUS is an efficient imaging modality that can detect pulmonary interstitial involvement in patients with systemic connective tissue disease with a high sensitivity and specificity. Further prospective studies conducted on a larger population are deemed necessary.

scholarly journals Evaluation of tubal patency by sonosalpingography is as good as hysterosalpingography in infertile women

2017 ◽  
Vol 6 (11) ◽  
pp. 5129
Author(s):  
C. Santhana Lakshmi ◽  
Kumara Sampath ◽  
Parvatha Vardani
Keyword(s):  
Sensitivity And Specificity ◽  
Predictive Value ◽  
High Sensitivity ◽  
Tubal Patency ◽  
Less Invasive ◽  
Significant Difference ◽  
Invasive Method ◽  
Tubal Factor ◽  
Primary Health Care Centers ◽  
Health Care Centers

Background: Tubal pathology is one of the causes of infertility in sub fertile couples. Therefore, evaluation of fallopian tubes forms an essential part of evaluation in a sub fertile woman. The aim of the study was to compare the efficacy of sonosalpingography which is a less invasive method when compared to hysterosalpingogram for evaluation of utero-tubal factor in females.Methods: A total of 95 patients 69 with primary infertility and 26 with secondary infertility attending our Obstetrics and Gynecology department were recruited for the study from June 2016 to July 2017. All the study subjects underwent Sonosalpingography on day 5th to 7th and Hysterosalpingography on 7th to 9th day of the menstrual cycle. Data was collected and compared to assess the sensitivity and specificity of sonosalpingography over hysterosalpingography. Statistical analysis was done by using SPSS software version 16.0.Results: Sonosalpingography (SSG) has 97% sensitivity and 94% specificity in comparison to hysterosalpingography (HSG). Analysis of the raw data gave positive predictive value of 98.3% and negative predictive value of 75%. From the results of this study it clear that there is no statistically significant difference (p = 0.237) between the results of the two methods (SSG and HSG).Conclusions: As sonosalpingogrpahy has high sensitivity and specificity and is less invasive. It should be used initially to assess tubal patency in cases of infertility. SSG is found to be a reliable, relatively less expensive diagnostic and therapeutic procedure in the management of infertility in females. Easy availability and accessibility of ultrasound in all primary health care centers definitely can prove superior to conventional method of diagnosis of tubal patency in such patient.

scholarly journals Assessing the Role of Uric Acid as a Predictor of Preeclampsia

2022 ◽  
Vol 12 ◽  
Author(s):  
Ana I. Corominas ◽  
Yollyseth Medina ◽  
Silvia Balconi ◽  
Roberto Casale ◽  
Mariana Farina ◽  
...  
Keyword(s):  
Uric Acid ◽  
Serum Uric Acid ◽  
Predictive Value ◽  
Gestational Hypertension ◽  
Low Cost ◽  
High Sensitivity ◽  
Use Of Resources ◽  
Before And After ◽  
High Negative Predictive Value ◽  
Group A

We assessed the diagnostic utility of uric acid for the prediction of preeclampsia. An observational prospective approach was carried out during 2014. Preeclamptic women were classified into 4 groups accordingly to the onset of preeclampsia and the presence of intrauterine growth restriction (IUGR). Serum uric acid levels, urea, and creatinine were measured. Receiver operating curves (ROC) of the uric acid levels ratio (UAr) between a dosage before and after the 20th week of gestation were performed. One thousand two hundred and ninety-third pregnant women were enrolled in this study. Eight hundred ten had non-complicated pregnancies, 40 preeclampsia, 33 gestational hypertension, and 20 IUGR without preeclampsia. Uric acid significantly raised after 20 weeks of gestation in women who develop preeclampsia before 34 weeks (Group A) or in those who develop preeclampsia after 37 weeks associated with IUGR (Group C). In women who develop preeclampsia after 34 weeks without IUGR (Groups B and D), uric acid increased after the 30th week of gestation. In all groups, UAr was greater than 1.5. In gestational hypertension, UAr was superior to 1.5 toward the end of gestation, while in IUGR without preeclampsia, the behavior of serum uric acid was similar to non-complicated pregnancies. In all cases, urea and creatinine showed normal values, confirming that patients had no renal compromise. ROC area was 0.918 [95% confidence interval (CI): 0.858–0.979) for the preeclampsia group and 0.955 (95% CI: 0.908–1.000) for Group A. UAr at a cut-off point ≥1.5 had a very low positive predictive value, but a high negative predictive value of 99.5% for preeclampsia and it reached 100% for Group A. Thus, a UAr less than 1.5 may be a helpful parameter with a strong exclusion value and high sensitivity for those women who are not expected to develop preeclampsia. Additionally, this low-cost test would allow for better use of resources in developing countries.

scholarly journals Rapid Detection of Cysticercus cellulosae by an Up-Converting Phosphor Technology-Based Lateral-Flow Assay

2021 ◽  
Vol 11 ◽  
Author(s):  
Dejia Zhang ◽  
Yu Qi ◽  
Yaxuan Cui ◽  
Weiyi Song ◽  
Xinrui Wang ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Rapid Detection ◽  
Small Volume ◽  
Taenia Solium ◽  
High Sensitivity ◽  
Lateral Flow ◽  
Neglected Tropical Disease ◽  
Porcine Cysticercosis ◽  
Cysticercus Cellulosae ◽  
Higher Sensitivity

Cysticercosis is a neglected tropical disease caused by the larvae of Taenia solium in pigs and humans. The current diagnosis of porcine cysticercosis is difficult, and traditional pathological tests cannot meet the needs of detection. This study established a UPT-LF assay for the detection of Cysticercus cellulosae. UCP particles were bound to two antigens, TSOL18 and GP50; samples were captured, and the signal from the UCP particles was converted into a detectable signal for analysis using a biosensor. Compared to ELISA, UPT-LF has higher sensitivity and specificity, with a sensitivity of 93.59% and 97.44%, respectively, in the case of TSOL18 and GP50 antigens and a specificity of 100% for both. Given its rapidness, small volume, high sensitivity and specificity, and good stability and reproducibility, this method could be used in the diagnosis of cysticercosis.

scholarly journals Evaluation of a Rapid Kit for Detection of IgM against Leptospira in Human

2019 ◽  
Vol 2019 ◽  
pp. 1-4 ◽  
Author(s):  
Mohan Rao ◽  
Fairuz Amran ◽  
Nadia Aqilla
Keyword(s):  
Sensitivity And Specificity ◽  
Predictive Value ◽  
Clinical Manifestations ◽  
Rapid Test ◽  
Febrile Illness ◽  
High Sensitivity ◽  
Confirmatory Test ◽  
Standard Laboratory ◽  
Test Efficiency ◽  
Sensitivity Specificity

Introduction. Leptospirosis is an acute febrile illness, known for its protean clinical manifestations and the challenge in differentiating from other infectious diseases. Standardized confirmatory test is antibody dependent and not accessible by the suburban community. This study measures efficiency of an immune-chromatographic assay, Leptocheck WB, in detecting acute leptospirosis. Methods. A total of 142 sera were used for kit evaluation. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were calculated by comparing rapid kit results with gold standard laboratory, microscopic agglutination test (MAT). Results. We found this rapid kit to have a sensitivity and specificity of 66.6% and 78.9%, respectively, whereas the PPV and NPV of the kit appeared to be 73.3% and 73.2%, respectively. Discussion. Test efficiency of this rapid kit is reasonable. It is specific in detecting leptospiral antibody and assures clinician of accurate diagnosis by having higher PPV and NPV. It is prompt and efficient in comparison with conventional methods in assisting differential diagnosis. High sensitivity and specificity leptospirosis rapid test is indeed a crucial measure to assist the diagnosis of acute undifferentiated febrile illnesses.

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