scholarly journals Improved Immunodiagnosis of Cystic Hydatid Disease by Using a Synthetic Peptide with Higher Diagnostic Value Than That of Its Parent Protein, Echinococcus granulosus Antigen B

2000 ◽  
Vol 38 (11) ◽  
pp. 3979-3983 ◽  
Author(s):  
Gualberto González-Sapienza ◽  
Carmen Lorenzo ◽  
Alberto Nieto
Keyword(s):  
Hydatid Disease ◽  
Synthetic Peptides ◽  
Diagnostic Value ◽  
Diagnostic Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Cystic Hydatid Disease ◽  
Antigen B ◽  
Parent Protein ◽  
Diagnostic Sensitivity And Specificity

The assays are used for the diagnosis of hydatid disease are still imperfect. The reported diagnostic sensitivity and specificity vary greatly depending on the panel of sera used, the laboratory conducting the assay, and, more critically, the antigen used. To contribute to its standardization, we have recently ranked the diagnostic performances of the major parasite antigens and the available synthetic peptides using a large collection of serum samples. That work showed that antigen B (AgB) possesses the highest diagnostic value among these antigens. In the present work we further dissected its antigenicity by analyzing the reactivity of the same panel of sera against a set of synthetic peptides spanning the sequence of both AgB subunits. The N-terminal extension of these subunits appeared to be immunodominant in human infections. A 38-mer peptide (p176) delineated from the N-terminal extension of the AgB/1 subunit performed in an enzyme-linked immunosorbent assay with a higher diagnostic sensitivity (80%) and specificity (94%) than native AgB, Ag5, or any other peptide antigen tested against this collection of serum samples. In view of its high diagnostic value and its nature as a well-defined reproducible antigen, p176 could conveniently be used as a reference standard antigen in the diagnosis of hydatid disease.

scholarly journals A Novel Enzyme-Linked Immunosorbent Assay for Diagnosis of Mycobacterium avium subsp. paratuberculosis Infections (Johne's Disease) in Cattle

2006 ◽  
Vol 13 (5) ◽  
pp. 535-540 ◽  
Author(s):  
C. A. Speer ◽  
M. Cathy Scott ◽  
John P. Bannantine ◽  
W. Ray Waters ◽  
Yasuyuki Mori ◽  
...  
Keyword(s):  
Sensitivity And Specificity ◽  
Mycobacterium Avium ◽  
Johne's Disease ◽  
Johne’S Disease ◽  
Diagnostic Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Formaldehyde Concentration ◽  
Serum Samples ◽  
Mycobacterium Avium Subsp Paratuberculosis ◽  
Diagnostic Sensitivity And Specificity

ABSTRACT Enzyme-linked immunosorbent assays (ELISAs) for the diagnosis of Johne's disease (JD), caused by Mycobacterium avium subsp. paratuberculosis, were developed using whole bacilli treated with formaldehyde (called WELISA) or surface antigens obtained by treatment of M. avium subsp. paratuberculosis bacilli with formaldehyde and then brief sonication (called SELISA). ELISA plates were coated with either whole bacilli or sonicated antigens and tested for reactivity against serum obtained from JD-positive and JD-negative cattle or from calves experimentally inoculated with M. avium subsp. paratuberculosis, Mycobacterium avium subsp. avium, or Mycobacterium bovis. Because the initial results obtained from the WELISA and SELISA were similar, most of the subsequent experiments reported herein were performed using the SELISA method. To optimize the SELISA test, various concentrations (3.7 to 37%) of formaldehyde and intervals of sonication (2 to 300 s) were tested. With an increase in formaldehyde concentration and a decreased interval of sonication, there was a concomitant decrease in nonspecific binding by the SELISA. SELISAs prepared by treating M. avium subsp. paratuberculosis with 37% formaldehyde and then a 2-s burst of sonication produced the greatest difference (7×) between M. avium subsp. paratuberculosis-negative and M. avium subsp. paratuberculosis-positive serum samples. The diagnostic sensitivity and specificity for JD by the SELISA were greater than 95%. The SELISA showed subspecies-specific detection of M. avium subsp. paratuberculosis infections in calves experimentally inoculated with M. avium subsp. paratuberculosis or other mycobacteria. Based on diagnostic sensitivity and specificity, the SELISA appears superior to the commercial ELISAs routinely used for the diagnosis of JD.

SOX Antibodies in Small-Cell Lung Cancer and Lambert-Eaton Myasthenic Syndrome: Frequency and Relation With Survival

2009 ◽  
Vol 27 (26) ◽  
pp. 4260-4267 ◽  
Author(s):  
Maarten J. Titulaer ◽  
Rinse Klooster ◽  
Marko Potman ◽  
Lidia Sabater ◽  
Francesc Graus ◽  
...  
Keyword(s):  
Western Blot ◽  
High Throughput Screening ◽  
Small Cell Lung Carcinoma ◽  
Diagnostic Value ◽  
Small Cell ◽  
Enzyme Linked Immunosorbent Assay ◽  
Small Cell Lung ◽  
Serum Samples ◽  
Cell Lung Carcinoma ◽  
Myasthenic Syndrome

Purpose SOX1 antibodies are common in small-cell lung carcinoma (SCLC) with and without paraneoplastic syndrome (PNS) and can serve as serological tumor marker. Addition of other antibodies might improve its diagnostic power. We validated an enzyme-linked immunosorbent assay (ELISA) to assess the diagnostic value of serum antibodies in SCLC and Lambert-Eaton myasthenic syndrome (LEMS). Clinical outcome with respect to SOX antibodies was evaluated, as the SOX-related antitumor immune response might help to control the tumor growth. Patients and Methods We used recombinant SOX1, SOX2, SOX3, SOX21, HuC, HuD, or HelN1 proteins in an ELISA to titrate serum samples and validated the assay by western blot. We tested 136 consecutive SCLC patients, 86 LEMS patients (43 with SCLC), 14 patients with SCLC and PNS (paraneoplastic cerebellar degeneration or Hu syndrome), 62 polyneuropathy patients, and 18 healthy controls. Results Our ELISA was equally reliable as western blot. Forty-three percent of SCLC patients and 67% of SCLC-LEMS patients had antibodies to one of the SOX or Hu proteins. SOX antibodies had a sensitivity of 67% and a specificity of 95% to discriminate between LEMS with SCLC and nontumor LEMS. No difference in survival was observed between SOX positive and SOX negative SCLC patients. Conclusion SOX antibodies are specific serological markers for SCLC. Our assay is suitable for high throughput screening, detecting 43% of SCLC. SOX antibodies have diagnostic value in discriminating SCLC-LEMS from nontumor LEMS, but have no relation to survival in patients with SCLC.

scholarly journals Acute Aortic Dissection Biomarkers Identified Using Isobaric Tags for Relative and Absolute Quantitation

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Ziya Xiao ◽  
Yuan Xue ◽  
Chenling Yao ◽  
Guorong Gu ◽  
Yaping Zhang ◽  
...  
Keyword(s):  
Aortic Dissection ◽  
Sensitivity And Specificity ◽  
Acute Aortic Dissection ◽  
Roc Curves ◽  
Diagnostic Value ◽  
Serum Samples ◽  
Absolute Quantitation ◽  
Isobaric Tags ◽  
Diagnostic Sensitivity And Specificity ◽  
D Dimer

The purpose of this study was to evaluate the utility of potential serum biomarkers for acute aortic dissection (AAD) that were identified by isobaric Tags for Relative and Absolute Quantitation (iTRAQ) approaches. Serum samples from 20 AAD patients and 20 healthy volunteers were analyzed using iTRAQ technology. Protein validation was performed using samples from 120 patients with chest pain. A total of 355 proteins were identified with the iTRAQ approach; 164 proteins reached the strict quantitative standard, and 125 proteins were increased or decreased more than 1.2-fold (64 and 61 proteins were up- and downregulated, resp.). Lumican, C-reactive protein (CRP), thrombospondin-1 (TSP-1), and D-dimer were selected as candidate biomarkers for the validation tests. Receiver operating characteristic (ROC) curves show that Lumican and D-dimer have diagnostic value (area under the curves [AUCs] 0.895 and 0.891,P<0.05). For Lumican, the diagnostic sensitivity and specificity were 73.33% and 98.33%, while the corresponding values for D-dimer were 93.33% and 68.33%. For Lumican and D-dimer AAD combined diagnosis, the sensitivity and specificity were 88.33% and 95%, respectively. In conclusion, Lumican has good specificity and D-dimer has good sensitivity for the diagnosis of AAD, while the combined detection of D-dimer and Lumican has better diagnostic value.

Seroprevalence of Human Cystic Echinococcosis in Sanandaj City, Kurdistan Province, Western Iran

2021 ◽  
Author(s):  
Naser Nazari ◽  
Tooran Nayeri ◽  
Farkhondeh Hazrati
Keyword(s):  
Sex Education ◽  
Cystic Echinococcosis ◽  
Elisa Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Cluster Sampling ◽  
Serum Samples ◽  
Cystic Hydatid Disease ◽  
Western Iran ◽  
Significant Difference ◽  
Human Cystic Echinococcosis

Background and Aims: Echinococcus granulosus (E. granulosus) is a cestode parasite that causes cystic hydatid disease in humans worldwide. Iran is one of the endemic regions for infection that indicate the importance and presence of infection in this country. Therefore, the current research aimed to characterize the seroprevalence of human cystic echinococcosis in Sanandaj city, Kurdistan province, western Iran. Materials and methods: Totally, 500 serum samples were collected from patients referred to different health centers in Sanandaj city using cluster sampling in 2018-2019. All the sera were examined using the enzyme-linked immunosorbent assay test. Results: The seroprevalence of human hydatidosis was reported at 2.2% by ELISA test in Sanandaj city. This rate was 9 (1.9%) in women and 2 (0.4) in men. The age group of 20-30 years old had the highest positivity rate (1.0%). Also, the subjects that consumed home slaughtered meat had the highest infection rate at 4 (0.8%). There was no significant difference regarding factors studied such as sex, education, residence, consumed water, keeping a dog, and the seropositivity. Conclusions: Seroprevalence of human cystic echinococcosis in Sanandaj city is lower than the general prevalence in Iran. Our research team hopes to provide accurate data on the prevalence of hydatidosis in Sanandaj encourage more extensive research to help prevent this parasite in Iran and worldwide.

scholarly journals Development of a Competitive Enzyme-Linked Immunosorbent Assay for Detection of Antibodies against the 3B Protein of Foot-and-Mouth Disease Virus

2015 ◽  
Vol 22 (4) ◽  
pp. 389-397 ◽  
Author(s):  
Ming Yang ◽  
Satya Parida ◽  
Tim Salo ◽  
Kate Hole ◽  
Lauro Velazquez-Salinas ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Diagnostic Sensitivity ◽  
Antibody Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Nonstructural Proteins ◽  
Serum Samples ◽  
Mouth Disease Virus ◽  
Foot And Mouth

ABSTRACTFoot-and-mouth disease (FMD) is one of the most highly contagious and economically devastating diseases, and it severely constrains the international trade of animals. Vaccination against FMD is a key element in the control of FMD. However, vaccination of susceptible animals raises critical issues, such as the differentiation of infected animals from vaccinated animals. The current study developed a reliable and rapid test to detect antibodies against the conserved, nonstructural proteins (NSPs) of the FMD virus (FMDV) to distinguish infected animals from vaccinated animals. A monoclonal antibody (MAb) against the FMDV NSP 3B was produced. A competitive enzyme-linked immunosorbent assay (cELISA) for FMDV/NSP antibody detection was developed using a recombinant 3ABC protein as the antigen and the 3B-specific MAb. Sera collected from naive, FMDV experimentally infected, vaccinated carrier, and noncarrier animals were tested using the 3B cELISA. The diagnostic specificity was 99.4% for naive animals (cattle, pigs, and sheep) and 99.7% for vaccinated noncarrier animals. The diagnostic sensitivity was 100% for experimentally inoculated animals and 64% for vaccinated carrier animals. The performance of this 3B cELISA was compared to that of four commercial ELISA kits using a panel of serum samples established by the World Reference Laboratory for FMD at The Pirbright Institute, Pirbright, United Kingdom. The diagnostic sensitivity of the 3B cELISA for the panel of FMDV/NSP-positive bovine serum samples was 94%, which was comparable to or better than that of the commercially available NSP antibody detection kits. This 3B cELISA is a simple, reliable test to detect antibodies against FMDV nonstructural proteins.

scholarly journals Chemiluminescence Immunoassay for the Detection of Antibodies against the 2C and 3ABC Nonstructural Proteins Induced by Infecting Pigs with Foot-and-Mouth Disease Virus

2017 ◽  
Vol 24 (8) ◽  
Author(s):  
Zezhong Liu ◽  
Junjun Shao ◽  
Furong Zhao ◽  
Guangqing Zhou ◽  
Shandian Gao ◽  
...  
Keyword(s):  
Disease Virus ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Diagnostic Sensitivity ◽  
High Rate ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Specificity ◽  
Serum Samples ◽  
Mouth Disease Virus ◽  
Foot And Mouth

ABSTRACT The potential diagnostic value of chemiluminescence immunoassays (CLIAs) has been accepted in recent years, although their use for foot-and-mouth disease (FMD) diagnostics has not been reported. Full-length 3ABC and 2C proteins were expressed in bacteria and purified by affinity chromatography to develop a rapid and accurate approach to distinguish pigs infected with foot-and-mouth disease virus (FMDV) from vaccinated pigs. The recombinant proteins were then used as antigens to develop two CLIAs for the detection of antibodies against nonstructural viral proteins. The diagnostic performance of the two assays was compared by analyzing serum from pigs (naive pigs, n = 63; vaccinated, uninfected pigs, n = 532; naive, infected pigs, n = 117) with a known infection status. The 3ABC-2C CLIA had a higher accuracy rate, with a diagnostic sensitivity of 100% and a diagnostic specificity of 96.5%, than the 3ABC CLIA, which had a diagnostic sensitivity of 95.7% and a diagnostic specificity of 96.0%. The results of the 3ABC-2C CLIA also had a high rate of concordance with those of two commercial FMDV enzyme-linked immunosorbent assay (ELISA) kits used to assess serum collected from 962 pigs in the field (96.2% and 97.8%, respectively). The 3ABC-2C CLIA detected infection in serum samples from infected pigs earlier than the commercial ELISA kits. In addition, the 3ABC-2C CLIA produced results within 15 min. On the basis of these findings, the 3ABC-2C CLIA could serve as the foundation for the development of penside FMD diagnostics and offers an alternative method to detect FMDV infections.

Three highly sensitive "bedside" serum and urine tests for pregnancy compared

1990 ◽  
Vol 36 (9) ◽  
pp. 1686-1688 ◽  
Author(s):  
H Christensen ◽  
H H Thyssen ◽  
O Schebye ◽  
A Berget
Keyword(s):  
Paired Comparison ◽  
False Negative ◽  
Diagnostic Sensitivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Negative Results ◽  
False Negative Results ◽  
Significant Difference ◽  
Comparison Of The Results ◽  
Urine And Serum

Abstract We examined three enzyme-linked immunosorbent assay (ELISA) kits for human choriogonadotropin (hCG) (pregnancy tests) for use with urine and serum samples: the Tandem Icon II hCG Urine and Tandem Icon II hCG Serum, the NovoClone Target hCG Test, and the Abbott TestPacks hCG-urine and hCG-serum. Paired comparison of the results from each kit indicated that the NovoClone Target assay showed significantly lower diagnostic sensitivity (P less than 0.05) than did the Tandem Icon II or Abbott TestPack, both for urine and for serum samples. None of the products demonstrated any significant difference (P greater than 0.05) in diagnostic specificity, but the NovoClone Target kit showed several serious false-negative results with both urine and serum. Paired testing of urine kits vs serum kits also showed no significant differences (P greater than 0.05) in diagnostic sensitivity or specificity. We found the Abbott kits to be the most convenient to use and to read.

Assessment of monoclonal antibodies to Echinococcus granulosus Antigen 5 and Antigen B for detection of human hydatid circulating antigens

Parasitology ◽  
1993 ◽  
Vol 106 (1) ◽  
pp. 75-81 ◽  
Author(s):  
D. Liu ◽  
M. D. Rickard ◽  
M. W. Lightowlers
Keyword(s):  
Monoclonal Antibodies ◽  
Echinococcus Granulosus ◽  
Binding Capacity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Antigen B ◽  
Detection Assay ◽  
Circulating Antigens ◽  
Normal Human ◽  
Antigen 5

SUMMARYFour monoclonal antibodies (MAb) to Echinococcus granulosus Antigen 5 (Ag5) and Antigen B (AgB) were assessed in an enzyme-linked immunosorbent assay (ELISA) for detection of circulating antigens (CAg) in sera of human patients with E. granulosus infection. Around 5·5–8% of 200 sera from 42 surgically proven hydatid patients contained detectable CAg by individual MAb. The combined detection rate for CAg, using four MAb, was 19% (38/200). Although hydatid CAg was detected by MAb in at least one serum sample from 21 of 42 patients, some patients remained negative in the assay regardless of the time when serum samples were taken (pre- or post-operatively), or of the continuing presence of hydatid cysts, their location or fertility. In addition, it was observed that the binding capacity of MAb for sheep hydatid cyst fluid antigen (SHCF) was somewhat reduced in the presence of normal human serum. The CAg detection assay would only be useful for assessment of hydatid infection status in patients with detectable CAg in serum samples.

scholarly journals Serological Diagnosis of Lung Cystic Hydatid Disease Using the Synthetic p176 Peptide

2012 ◽  
Vol 19 (6) ◽  
pp. 944-947 ◽  
Author(s):  
Saul J. Santivañez ◽  
Patricia Arias ◽  
Milagrytos Portocarrero ◽  
Silvia Rodriguez ◽  
Armando E. Gonzalez ◽  
...  
Keyword(s):  
Hydatid Disease ◽  
Synthetic Peptide ◽  
Serum Antibody ◽  
Multivariate Logistic Regression Analysis ◽  
Case Series ◽  
Diagnostic Tools ◽  
Enzyme Linked Immunosorbent Assay ◽  
X Rays ◽  
Cystic Hydatid Disease ◽  
Cystic Hydatid

ABSTRACTCystic hydatid disease (CHD) is a worldwide zoonosis caused by the larval stage of the dog tapewormEchinococcus granulosus. Diagnosis is based on imagenological tools (abdominal ultrasound, chest X-rays, or computed tomography [CT] scan). Serological antibody-detecting assays, using diverse native antigens, have been used as a supportive diagnostic tool, but their sensitivities and specificities differ greatly. The use of synthetic peptides as antigens should provide more reliability and allow better assessment and comparison of test formats and case series. The synthetic peptide p176, corresponding to the N-terminal extreme of the subunit of antigen B (AgB8/1), has shown promising performances for diagnosis of CHD. We evaluated the performance of the synthetic peptide p176 for the diagnosis of pulmonary hydatid disease in an enzyme-linked immunosorbent assay (ELISA) format. Sixty-one serum samples from patients with a diagnosis of pulmonary hydatidosis confirmed by surgery and 128 from healthy volunteers were tested. The overall sensitivity and specificity of the p176 ELISA for lung CHD were 78.69% and 96.88%, respectively. On bivariate analysis, positive serum antibody reactions were associated with the presence of complications and with the number of cysts (single/multiple). Only the presence of persistent complications significantly associated with seropositivity on multivariate logistic regression analysis (odds ratio [OR], 9.58; 95% confidence interval [CI], 2.15 to 42.6;P= 0.003). The p176 ELISA performs well for the diagnosis of lung CHD and adds an easily reproducible diagnostic assay to the existing diagnostic tools.

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