scholarly journals Comparison of Western Blot Technique with Microscopy and Culture for Diagnosis of Tuberculosis

2016 ◽  
Vol 1 (1) ◽  
pp. 21-24
Author(s):  
Md. Jamal Uddin Gazi ◽  
Md. Ruhul Amin Miah ◽  
Sharmeen Ahmed ◽  
Abu Naser Ibne Sattar
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Antibody Response ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Polyacrylamide Gel ◽  
Serum Samples ◽  
Bacteriological Diagnosis ◽  
Blot Technique ◽  
Study Population

Diagnosis of tuberculosis is usually done by microscopy for AFB and other tests. But each test has different limitation. This study was carried out to compare western blot technique with microscopy and culture for diagnosis of tuberculosis. Sputum and other samples were collected from 112 TB patients for bacteriological diagnosis by microscopy and culture for AFB. Serum samples were collected from 112 TB patients and 50 control subjects for detection of antibody response by western blot. For western blot analysis, Mycobacterium tuberculosis sonicate antigen extract was fractionated by electrophoresis on  polyacrylamide gel.  Western blot analysis revealed four polypeptides against which antibody response of study population were observed. The sensitivity of western blot was 73.21% which was higher than that of microscopy (63.39%) and culture (57.14%) for diagnosis of tuberculosis. The specificity of western blot was 92%.Bangladesh J Med Microbiol 2007; 01 (01):21-24

Lav/HTLV-III Neutralizing Antibodies in the Sera of Patients with Aids, Lymphadenopathy Syndrome and Asymptomatic Seropositive Individuals

1986 ◽  
Vol 72 (3) ◽  
pp. 219-224 ◽  
Author(s):  
Georgine P. Faulkner-Valle ◽  
Anita De Rossi ◽  
Oriella Dalla Gassa ◽  
Luigi Chieco-Bianchi
Keyword(s):  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Neutralizing Antibodies ◽  
Elisa Test ◽  
Antibody Specificity ◽  
Serum Samples ◽  
Neutralizing Activity ◽  
The Mean ◽  
Asymptomatic Individuals

Serum samples which had previously been found positive for LAV/HTLV-III antibodies by the ELISA test and then confirmed by radioimmunoassay (Western blot) were tested for the presence of neutralizing antibodies. No neutralizing activity was found in the sera of a group of patients with the clinical diagnosis of AIDS. However in patients with LAS and other related pathologic conditions the percentage of sera positive for neutralizing antibodies was 27 % and 55 % respectively. At least 50 % of the sera from seropositive asymptomatic individuals had neutralizing activity but with the exception of the haemophiliac group the mean titre was much lower than that of LAS patients. No relationship was found between the neutralizing titre and the antibody specificity detected by Western blot analysis for p41 and p120.

scholarly journals Western Blot Analysis of the IgG-Antibody Response to Acid- Glycine-Extracted Antigens from Campylobacter fetus subsp. fetus and C. jejuni in Naturally Infected Sheep

2007 ◽  
Vol 76 (2) ◽  
pp. 245-251 ◽  
Author(s):  
K. Gürtürk ◽  
I. H. Ekin ◽  
A. Arslan
Keyword(s):  
Antibody Response ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Igg Antibodies ◽  
Igg Antibody ◽  
Serological Tests ◽  
Recent Infection ◽  
Healthy Sheep ◽  
Apparently Healthy

IgG-antibody response in aborting sheep and in apparently healthy sheep in a flock against acidglycine- extracted antigens from three strains for each C. fetus subsp. fetus and C. jejuni were analysed by Western blot. One strain of C. fetus subsp. fetus was isolated from aborting sheep. Western blot analysis of the sera revealed the presence of IgG antibody binding to the common antigens including proteins with the Mw of 63 kDa and 54 kDa in extracts from both C. fetus subsp. fetus and C. jejuni strains. In addition, IgG antibodies in sera from aborting sheep reacted more strongly with the antigens from C. fetus subsp. fetus strains with Mw of approximately 100, 95 and 86.5 kDa than those of apparently healthy sheep. The binding profile of the antibodies with these antigens appeared to be unique for each C. fetus subsp. fetus strain. On the other hand, IgG antibodies only in sera from aborting sheep recognized strongly the antigens of each C. fetus subsp. fetus strain at the Mw ranged from approximately 26 to 22 kDa. However, the antigenic components between 26 and 22 kDa were not detectable in coomassie blue stained gel and thought to have non-protein nature. These low molecular weight antigens of C. fetus subsp. fetus may be related to a recent infection in aborting sheep. These observations indicate that such speciesspecific antigens or conjugated protein antigens could be used for improving the specificity of the serological tests to detect C. fetus antibodies in sheep sera, and may be the candidates for subunit vaccines against ovine abortion.

scholarly journals Time-Dependent Degradation Pattern of Cardiac Troponin T Following Myocardial Infarction

2013 ◽  
Vol 59 (7) ◽  
pp. 1083-1090 ◽  
Author(s):  
Eline PM Cardinaels ◽  
Alma MA Mingels ◽  
Tom van Rooij ◽  
Paul O Collinson ◽  
Frits W Prinzen ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Western Blot ◽  
Blot Analysis ◽  
Cardiac Troponin ◽  
Western Blot Analysis ◽  
Troponin T ◽  
Time Dependent ◽  
Cardiac Troponin T ◽  
Serum Samples ◽  
Degradation Pattern

BACKGROUNDCardiac troponin T (cTnT) is widely used for the diagnosis of acute myocardial infarction (AMI). However, it is still unclear whether degraded cTnT forms circulate in the patient's blood. We therefore aimed to elucidate which cTnT forms are detected by the clinical assay.METHODSSeparation of cTnT forms by gel filtration chromatography (GFC) was performed in sera from 13 AMI patients to examine cTnT degradation. The GFC eluates were subjected to Western blot analysis with the original antibodies from the Roche immunoassay used to mimic the clinical cTnT assay. To investigate the degradation pattern with time, standardized serum samples of 18 AMI patients collected 0–72 h after admission were analyzed by Western blot analysis.RESULTSGFC analysis of AMI patients' sera revealed 2 cTnT peaks with retention volumes of 5 and 21 mL. Western blot analysis identified these peaks as cTnT fragments of 29 and 14–18 kDa, respectively. Furthermore, the performance of direct Western blots on standardized serum samples demonstrated a time-dependent degradation pattern of cTnT, with fragments ranging between 14 and 40 kDa. Intact cTnT (40 kDa) was present in only 3 patients within the first 8 h after hospital admission.CONCLUSIONSThese results demonstrate that the Roche cTnT immunoassay detects intact as well as degraded cTnT forms in AMI patients' sera during the period of diagnostic testing. Moreover, following AMI, cTnT is degraded in a time-dependent pattern.

scholarly journals Subcellular Localization of the Intracellular Survival-Enhancing Eis Protein of Mycobacterium tuberculosis

2001 ◽  
Vol 69 (7) ◽  
pp. 4295-4302 ◽  
Author(s):  
John L. Dahl ◽  
Jun Wei ◽  
James W. Moulder ◽  
Suman Laal ◽  
Richard L. Friedman
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Amino Acid ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Intracellular Survival ◽  
Terminal Amino Acid ◽  
Terminal Amino

ABSTRACT Mycobacterium tuberculosis is a facultative intracellular pathogen that has evolved the ability to survive and multiply within human macrophages. It is not clear how M. tuberculosis avoids the destructive action of macrophages, but this ability is fundamental in the pathogenicity of tuberculosis. A gene previously identified in M. tuberculosis, designatedeis, was found to enhance intracellular survival ofMycobacterium smegmatis in the human macrophage-like cell line U-937 (J. Wei et al., J. Bacteriol. 182:377–384, 2000). Wheneis was introduced into M. smegmatis on a multicopy vector, sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the appearance of a unique 42-kDa protein band corresponding to the predicted molecular weight of the eisgene product. This band was electroeluted from the gel with a purity of >90% and subjected to N-terminal amino acid sequencing, which demonstrated that the 42-kDa band was indeed the protein product ofeis. The Eis protein produced by M. tuberculosis H37Ra had an identical N-terminal amino acid sequence. A synthetic polypeptide corresponding to a carboxyl-terminal region of the deduced eis protein sequence was used to generate affinity-purified rabbit polyclonal antibodies that reacted with the 42-kDa protein in Western blot analysis. Hydropathy profile analysis showed the Eis protein to be predominantly hydrophilic with a potential hydrophobic amino terminus. Phase separation of M. tuberculosis H37Ra lysates by the nonionic detergent Triton X-114 revealed the Eis protein in both the aqueous and detergent phases. After fractionation of M. tuberculosis by differential centrifugation, Eis protein appeared mainly in the cytoplasmic fraction but also in the membrane, cell wall, and culture supernatant fractions as well. Forty percent of the sera from pulmonary tuberculosis patients tested for anti-Eis antibody gave positive reactions in Western blot analysis. Although the function of Eis remains unknown, evidence presented here suggests it associates with the cell surface and is released into the culture medium. It is produced during human tuberculosis infection and therefore may be an important M. tuberculosis immunogen.

scholarly journals Identification of novel serological autoantibodies in Takayasu arteritis patients using HuProt arrays

2020 ◽  
pp. mcp.RA120.002119
Author(s):  
Xiao-Ting Wen ◽  
Guang Song ◽  
Chao-Jun Hu ◽  
Jianbo Pan ◽  
Zi-Yan Wu ◽  
...  
Keyword(s):  
Autoimmune Disease ◽  
Western Blot ◽  
Phase Ii ◽  
Takayasu Arteritis ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Healthy Subjects ◽  
Serum Samples ◽  
Tree Model ◽  
Two Phase

To identify novel autoantibodies of Takayasu arteritis (TAK) using HuProt array-based approach. A two-phase approach was adopted. In Phase I, serum samples collected from 40 TAK patients, 15 autoimmune disease patients, and 20 healthy subjects were screened to identify TAK-specific autoantibodies using human protein (HuProt) arrays. In Phase II, the identified candidate autoantibodies were validated with TAK-focused arrays using an additional cohort comprised of 109 TAK patients, 110 autoimmune disease patients, and 96 healthy subjects. Subsequently, the TAK-specific autoantibodies validated in Phase II were further confirmed using Western blot analysis. We identified and validated eight autoantibodies as potential TAK-specific diagnostic biomarkers, including anti-SPATA7, -QDPR, -SLC25A2, -PRH2, -DIXDC1, -IL17RB, -ZFAND4, and -NOLC1 antibodies, with AUC of 0.803, 0.801, 0.780, 0.696, 0.695, 0.678, 0.635 and 0.613, respectively. SPATA7 could distinguish TAK from healthy and disease controls with 73.4% sensitivity at 85.4% specificity, while QDPR showed 71.6% sensitivity at 86.4% specificity. SLC25A22 showed the highest sensitivity of 80.7%, but at lower specificity of 67.0%. In addition, PRH2, IL17RB and NOLC1 showed good specificities of 88.3%, 85.9% and 86.9%, respectively, but at lower sensitivities (<50%). Finally, DIXDC1 and ZFAND4 showed moderate performance as compared with the other autoantibodies. Using a decision tree model, we could reach a specificity of 94.2% with AUC of 0.843, a significantly improved performance as compared to that by each individual biomarker. The performance of three autoantibodies, namely anti-SPATA7, -QDPR and -PRH2, were successfully confirmed with Western blot analysis. Using this two-phase strategy, we identified and validated eight novel autoantibodies as TAK–specific biomarker candidates, three of which could be readily adopted in a clinical setting.

scholarly journals Characterization of Immunoglobulin G and Its Subclass Response to Indian Kala-Azar Infection before and after Chemotherapy

2004 ◽  
Vol 72 (2) ◽  
pp. 863-870 ◽  
Author(s):  
Rajesh Ravindran ◽  
Khairul Anam ◽  
Bibhas C. Bairagi ◽  
Bibhuti Saha ◽  
Netai Pramanik ◽  
...  
Keyword(s):  
Western Blot ◽  
Immunoglobulin G ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Igg Subclasses ◽  
Cross Reaction ◽  
Future Application ◽  
Healthy Controls ◽  
Serum Samples ◽  
Kala Azar

ABSTRACT Serologic parameters of kala-azar were evaluated by Western blot analysis. Sera from kala-azar patients with confirmed diagnoses were screened for immunoglobulin G (IgG) and IgG subclass-specific reactivity against Leishmania donovani membrane antigen (LAg). Heterogenous LAg-specific IgG reactivity with numerous proteins with molecular masses ranging from 18 to 190 kDa was observed. Though the individual band patterns were varied, seven polypeptides of approximately 31, 34, 51, 63, 72, 91, and 120 kDa were immunoreactive with all the sera tested from kala-azar patients. The band patterns of the immunoblots of sera from patients after treatment and clinical cure with sodium antimony gluconate revealed a decrease in the frequency of the bands. Still, recognition of the 63- and 120-kDa bands was 100%, and the 55- and 91-kDa fractions were recognized in 93% of the sera from cured individuals. Among the IgG subclasses, IgG1 reacted with the greatest number of polypeptides. The 63-kDa protein was again detected by all of the IgG subclasses of all the sera tested. Other fractions recognized by the subclasses of more than 70% of the serum samples included those of 47, 51, 55, and 78 kDa. Following treatment, 63- and 51-kDa bands were the most reactive with the IgG subclasses. LAg-associated cross-reaction with other reference human antisera revealed a mild reactivity of the 63-kDa polypeptide with some of the serum samples from leprosy, malaria, typhoid, tuberculosis, and healthy controls. Western blot analysis of LAg entrapped in liposomes, strong vaccine candidates against experimental visceral leishmaniasis, revealed a more restricted band pattern. The 63-kDa fraction revealed by all pre- and posttreatment sera showed almost negligible levels of cross-reaction with sera from patients with other diseases or from healthy controls. These observations provide insight into induced immunity during kala-azar infection for future application.

Western blot analysis of antibody response to pneumococcal protein antigens in a murine model of pneumonia.

1997 ◽  
Vol 4 (6) ◽  
pp. 778-782
Author(s):  
H Mouneimne ◽  
M Juvin ◽  
J L Beretti ◽  
E Azoulay-Dupuis ◽  
E Vallee ◽  
...  
Keyword(s):  
Antibody Response ◽  
Western Blot ◽  
Murine Model ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Protein Antigens
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