A novel Western blot test containing both viral and recombinant proteins for anticytomegalovirus immunoglobulin M detection.

1997 ◽  
Vol 35 (2) ◽  
pp. 393-397 ◽  
Author(s):  
T Lazzarotto ◽  
G T Maine ◽  
P Dal Monte ◽  
A Ripalti ◽  
M P Landini
Keyword(s):  
Western Blot ◽  
Recombinant Proteins ◽  
Immunoglobulin M ◽  
Western Blot Test

A Preliminary Evaluation of a Prototype Western Blot Confirmatory Test Kit for Syphilis

1994 ◽  
Vol 5 (6) ◽  
pp. 409-414 ◽  
Author(s):  
H Young ◽  
P J Walker ◽  
D Merry ◽  
A Mifsud
Keyword(s):  
Western Blot ◽  
False Positive ◽  
High Sensitivity ◽  
High Specificity ◽  
Confirmatory Test ◽  
Preliminary Evaluation ◽  
Serological Tests ◽  
Western Blot Test ◽  
Specificity And Sensitivity ◽  
Test Kit

A prototype Western blot kit was evaluated as a confirmatory test for syphilis using 131 sera characterized by other serological tests for syphilis. There were 114 treponemal sera (including 94 cases of early syphilis, 83 of which were untreated) and 17 non-treponemal problem sera (11 gave false positive reactions on screening with the TmpA recombinant antigen enzyme immunoassay (EIA), 3 gave false positive fluorescent treponemal antibody absorbed (FTA-abs) tests, and 3 false positive Captia Syphilis G EIA results). Based on the manufacturer's criteria of reactivity in multiple bands for designating a positive result the Western blot test gave a sensitivity of 99.1% (113/114) and a specificity of 88.2% (15/17) when indeterminate reactions were scored positive and 98.2% (112/114) and 100% (17/17) when indeterminate reactions were scored negative. Sensitivity was high in both treated and untreated infection. Corresponding sensitivities for the TPHA and FTA-abs when equivocal reactions were scored negative were 97.5% (111/114) and 99.1% (113/114). The high sensitivity of the FTA-abs in this study is probably due to the large number of untreated primary infections. Our results with the Western blot, confirm earlier studies using ‘in-house’ test systems and, support a role for a commercial Western blot test in the confirmatory diagnosis of syphilis. Further studies are required to confirm the high specificity and sensitivity of the kit in a larger series including a wider variety of non-treponemal cases as well as patients with untreated and treated infection.

scholarly journals Human serum antibody reactivity towards Paracoccidioides brasiliensis antigens treated with sodium metaperiodate

2008 ◽  
Vol 41 (4) ◽  
pp. 325-329 ◽  
Author(s):  
Ana Paula Ferreira ◽  
Thadeu Côrrea ◽  
Rosângela Cunha ◽  
Marcos José Marques ◽  
Maria Angela Montesano ◽  
...  
Keyword(s):  
Western Blot ◽  
Human Serum ◽  
Paracoccidioides Brasiliensis ◽  
Serum Antibody ◽  
Parasitic Infections ◽  
Sodium Metaperiodate ◽  
Chronic Form ◽  
Western Blot Test ◽  
Immunoglobulin Isotypes ◽  
Acute Form

In this study, we evaluated the profile of anti-Paracoccidioides brasiliensis immunoglobulin isotypes in serum from patients with the acute and chronic forms of paracoccidioidomycosis, using the whole Paracoccidioides brasiliensis antigen and the antigen treated with sodium metaperiodate. All the immunoglobulin isotypes present in the serum from patients with the acute and chronic forms of paracoccidioidomycosis presented higher reactivity towards the whole antigen than to the antigen treated with metaperiodate (P < 0.05). The reactivity of IgG and IgM to the antigen treated with metaperiodate was greater in serum from patients with the acute form of the disease (P < 0.05), while IgA was more reactive in serum from patients with the chronic form (P < 0.05). There was greater reactivity of IgG1 and IgG2 to the whole antigen and the antigen treated with metaperiodate in the serum from patients with paracoccidioidomycosis than there was in serum from patients with other parasitic infections (P < 0.05). Furthermore, IgG1 from patients with the acute form recognized the 19kDa, 27kDa and 31kDa antigens in the western blot test. Thus, the results suggest that modifications to the epitopes of Paracoccidioides brasiliensis antigens may help to improve the immunodiagnosis of paracoccidioidomycosis.

scholarly journals Cloning and analysis of a recombinant antigen containing an epitope specific for human T-cell lymphotropic virus type II

Blood ◽  
1992 ◽  
Vol 79 (10) ◽  
pp. 2789-2796 ◽  
Author(s):  
KG Hadlock ◽  
JJ Lipka ◽  
TP Chow ◽  
SK Foung ◽  
GR Reyes
Keyword(s):  
Fusion Protein ◽  
Western Blot ◽  
Recombinant Proteins ◽  
Recombinant Antigen ◽  
Type Ii ◽  
Lambda Phage ◽  
Chain Reaction ◽  
Specific Epitope ◽  
Human T Cell ◽  
Polymerase Chain

Abstract An immunodominant HTLV-I-specific epitope in the HTLV-I envelope glycoprotein (GP) 46 has been described. To determine if the analogous region of HTLV-II contains a similarly immunogenic and specific epitope, the polymerase chain reaction (PCR) was used to amplify HTLV- II DNA fragments encoding various portions of the putative epitope. The synthesized DNAs were cloned into lambda-phage gt11 and screened for production of immunoreactive fusion protein using sera from HTLV-II- or HTLV-I-infected individuals. Antisera from HTLV-II-infected individuals identified three of four recombinant clones when tested in a plaque immunoassay. Fusion protein from one of the clones, GH2-K15, was purified and analyzed by Western blot against a panel of HTLV-I and HTLV-II antisera. Twenty-one of 22 HTLV-II-infected sera were reactive with the GH2-K15 epitope. Sera from HTLV-I-infected and HTLV-I- uninfected individuals did not cross-react with GH2-K15. Western blot analysis of recombinant proteins encoding portions of the HTLV-II sequences in the Gh2-K15 antigen localized the HTLV-II-specific epitope to a 17-amino acid sequence. Recombinant antigens containing this epitope should be useful for type-specific serologic diagnosis of HTLV- II infection.

scholarly journals Indirect ELISA for diagnosis of Brucella ovis infection in rams

2014 ◽  
Vol 66 (6) ◽  
pp. 1695-1702 ◽  
Author(s):  
S.A. França ◽  
J.P.S. Mol ◽  
E.A. Costa ◽  
A.P.C. Silva ◽  
M.N. Xavier ◽  
...  
Keyword(s):  
Western Blot ◽  
Recombinant Proteins ◽  
Indirect Elisa ◽  
High Sensitivity ◽  
Economic Losses ◽  
Well Testing ◽  
E Coli ◽  
Sheep Industry ◽  
Brucella Ovis ◽  
Sexually Mature

Brucella ovis is a major cause of epididymitis in sexually mature rams, resulting in subfertility, infertility, and economic losses for the sheep industry worldwide. The aim of this study was to develop an indirect ELISA (iELISA) using recombinant proteins, namely rBoP59 and rBP26, as antigens for serological diagnosis of B. ovisinfection. The BoP59 and BP26 recombinant proteins were expressed in E. coli and purified by affinity chromatography. Antigenicity was tested by Western blot and iELISA. Standardization of iELISA was performed with 500ng and 1µg BoP59 and rBP26 per well, testing serum from uninfected and experimentally infected rams. rBP26 was effective in distinguishing positive from negative rams. The rBP26 iELISA developed in this study is the first to use a completely purified rBP26 as antigen resulting in high sensitivity (100%) and specificity (90.2%), and an overall accuracy equal to 1.0.

scholarly journals Development of a Western Blot Assay for Detection of Antibodies against Coronavirus Causing Severe Acute Respiratory Syndrome

2004 ◽  
Vol 11 (2) ◽  
pp. 417-422 ◽  
Author(s):  
Qigai He ◽  
Kooi Hoong Chong ◽  
Hiok Hee Chng ◽  
Bernard Leung ◽  
Ai Ee Ling ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Western Blot ◽  
Recombinant Proteins ◽  
Immunofluorescence Assay ◽  
Sars Coronavirus ◽  
Western Blot Assay ◽  
Nucleocapsid Gene ◽  
Specificity And Sensitivity ◽  
C Terminus ◽  
Coronavirus Infection

ABSTRACT To identify a major antigenic determinant for use in the development of a rapid serological diagnostic test for severe acute respiratory syndrome (SARS) coronavirus infection and to study the immune response during SARS coronavirus infection in humans, we cloned the full length and six truncated fragments of the nucleocapsid gene, expressed them, and purified them as glutathione S-transferase-tagged recombinant proteins. The reactivities of the recombinant proteins to a panel of antibodies containing 33 SARS coronavirus-positive sera and 66 negative sera and to antibodies against other animal coronaviruses were screened. A truncated 195-amino-acid fragment from the C terminus of the nucleocapsid protein (N195) was identified that had a strong ability to detect antibodies against SARS coronavirus. No cross-reaction was found between the N195 protein and antibodies against chicken, pig, and canine coronaviruses. The N195 protein was used to develop a Western blot assay to detect antibodies against SARS coronavirus in 274 clinically blinded samples. The specificity and sensitivity of this test were 98.3 and 90.9%, respectively. The correlation between our Western blotting assay and an immunofluorescence assay (IFA) was also analyzed. The results of our Western blot assay and IFA for the detection of SARS coronavirus-positive sera were the same. Thus, the N195 protein was identified as a suitable protein to be used as an antigen in Western blot and other possible assays for the detection of SARS coronavirus infection.

Evaluation of a Western Blot Test, Helico Blot 2.1, in the Diagnosis of Helicobacter pylori Infection in a Pediatric Population

Helicobacter ◽  
2002 ◽  
Vol 7 (3) ◽  
pp. 210-215 ◽  
Author(s):  
Monica Oleastro ◽  
Rita Matos ◽  
Jose Cabral ◽  
Rosa Barros ◽  
Ana Isabel Lopes ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Western Blot ◽  
Pediatric Population ◽  
Helicobacter Pylori Infection ◽  
Western Blot Test
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