Analysis of Specific Immunoglobulin G Subclass Antibodies for Serological Diagnosis of Echinococcosis by a Standard Enzyme-Linked Immunosorbent Assay

ABSTRACT The potential roles of specific antibodies of the different immunoglobulin G (IgG) subclasses in the serological diagnosis of cystic echinococcosis (CE) and alveolar echinococcosis (AE) were investigated by an enzyme-linked immunosorbent assay based on hydatid fluid as antigen. Specific antibodies of subclass 1 were found to be of major importance. In sera collected at the time of diagnosis (i.e., before any therapeutic intervention was initiated) they could be demonstrated in 14 of 15 sera from patients with CE and in all 12 sera from patients with AE. The most discriminatory and the most specific antibodies found in this study belonged to IgG subclass 4. Only one false-positive reaction was observed with 253 sera from healthy volunteers, and no cross-reactions occurred in 80 sera from patients with different parasitic infections. Specific IgG4 antibodies could be demonstrated in 61.0 to 66.7% (CE) or 47.6 to 66.7% (AE) of the cases. Antibody levels of IgG subclass 2 were elevated only moderately, and subclass 3 antibodies were detected in a few cases only. In addition, nonspecific reactions in sera of healthy volunteers or patients with other parasitic infections could partially be attributed to antibodies of subclasses 2 and 3.

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Evaluation of immunoglobulin G4 subclass antibody in a peptide-based enzyme-linked immunosorbent assay for the serodiagnosis of human fascioliasis

Parasitology ◽

10.1017/s0031182007003435 ◽

2007 ◽

Vol 134 (14) ◽

pp. 2021-2026 ◽

Cited By ~ 12

Author(s):

C. TANTRAWATPAN ◽

W. MALEEWONG ◽

C. WONGKHAM ◽

S. WONGKHAM ◽

P. M. INTAPAN ◽

...

Keyword(s):

Immunosorbent Assay ◽

Large Scale ◽

Laboratory Diagnosis ◽

Enzyme Linked Immunosorbent Assay ◽

Parasitic Infections ◽

Immunoglobulin G4 ◽

Predictive Values ◽

Diagnostic Potential ◽

Human Fascioliasis ◽

Specific Igg4

SUMMARYTo improve the diagnosis of human fascioliasis caused byFasciola gigantica, we developed a peptide-based enzyme-linked immunosorbent assay (peptide-based ELISA) based on the detection of specific IgG4 subclass antibody. Two identified B-cell epitopes ofF. giganticacathepsin L1 were synthesized as single synthetic peptides, acetyl-DKIDWRESGYVTELKDQGNC-carboxamide (peptide L) and acetyl-DKIDWRESGYVTEVKDQGNC-carboxamide (peptide V), and their diagnostic potential was evaluated. The sera of 25 patients infected withF. gigantica, 212 patients with other parasitic infections, 32 cholangiocarcinoma patients and 57 healthy controls were analysed. The sensitivity, specificity, accuracy, and positive and negative predictive values of this assay were the same with both peptides at 100%, 99·7%, 99·7%, 96·2% and 100%, respectively. These highly sensitive and specific peptide-based ELISAs for the detection of specific IgG4 antibody could be useful for laboratory diagnosis of human fascioliasis in future large-scale surveys throughout Southeast Asia where this disease is prevalent.

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Heterogeneous Antibody Responses in Tuberculosis

Infection and Immunity ◽

10.1128/iai.66.8.3936-3940.1998 ◽

1998 ◽

Vol 66 (8) ◽

pp. 3936-3940 ◽

Cited By ~ 162

Author(s):

Konstantin Lyashchenko ◽

Roberto Colangeli ◽

Michel Houde ◽

Hamdan Al Jahdali ◽

Dick Menzies ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Antibody Response ◽

Immunosorbent Assay ◽

Immunoglobulin G ◽

Antigen Recognition ◽

Antibody Responses ◽

Enzyme Linked Immunosorbent Assay ◽

Protein Antigens ◽

Tuberculosis Patients ◽

Specific Antibodies

ABSTRACT Antibody responses during tuberculosis were analyzed by an enzyme-linked immunosorbent assay with a panel of 10 protein antigens of Mycobacterium tuberculosis. It was shown that serum immunoglobulin G antibodies were produced against a variety of M. tuberculosis antigens and that the vast majority of sera from tuberculosis patients contained antibodies against one or more M. tuberculosis antigens. The number and the species of serologically reactive antigens varied greatly from individual to individual. In a given serum, the level of specific antibodies also varied with the antigen irrespective of the total number of antigens recognized by that particular serum. These findings indicate that person-to-person heterogeneity of antigen recognition, rather than recognition of particular antigens, is a key attribute of the antibody response in tuberculosis.

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Rubella-specific IgG subclass avidity ELISA and its role in the differentiation between primary rubella and rubella reinfection

Epidemiology and Infection ◽

10.1017/s0950268800029459 ◽

1988 ◽

Vol 101 (3) ◽

pp. 591-598 ◽

Cited By ~ 55

Author(s):

H. I. J. Thomas ◽

P. Morgan-Capner

Keyword(s):

Optical Density ◽

Immunosorbent Assay ◽

Primary Infection ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Subclass ◽

Antibody Avidity ◽

Specific Antibodies ◽

Specific Igg ◽

Protein Denaturant

SUMMARYAn antiglobulin enzyme-linked immunosorbent assay for rubella-specific IgG1and IgG3was adapted to measure antibody avidity by incorporating a mild protein denaturant, diethylamine (DEA), into the serum diluent. Sera were tested at varying dilutions, both with and without DEA, if they contained sufficient specific IgG1or IgG3. The optical density (OD) was measured and curves were plotted. The highest OD (V) was noted and halved (V/2). The distance between the OD curves at V/2 was measured as the DEA shift value.Sera were examined from people whose sera contained rubella-specific antibodies as a consequence of infection or vaccination in the distant past (24 sera), recent primary rubella (66 sera), symptomatic reinfection (11 sera) or asymptomatic reinfection (64 sera). For specific IgG1the DEA shift value was <O·6 for cases of rubella in the distant past, compared with >0·8 for the first month after primary infection. The maximum DEA shift value for the sera from cases of reinfection was 0·65.No serum from cases of rubella in the distant past contained sufficient specific IgG3to estimate avidity. The sera collected within 1 month of onset of primary rubella gave DEA shift values >O·7 compared with sera from reinfections, which gave DEA shift values <O·6, except for two sera from a case of symptomatic reinfection.Thus the assessment of specific IgG subclass avidity is of value in differentiating serologically primary rubella from reinfection.

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Serological diagnosis of hantavirus infections by an enzyme-linked immunosorbent assay based on detection of immunoglobulin G and M responses to recombinant nucleocapsid proteins of five viral serotypes.

Journal of Clinical Microbiology ◽

10.1128/jcm.35.5.1122-1130.1997 ◽

1997 ◽

Vol 35 (5) ◽

pp. 1122-1130 ◽

Cited By ~ 75

Author(s):

F Elgh ◽

A Lundkvist ◽

O A Alexeyev ◽

H Stenlund ◽

T Avsic-Zupanc ◽

...

Keyword(s):

Immunosorbent Assay ◽

Immunoglobulin G ◽

Serological Diagnosis ◽

Enzyme Linked Immunosorbent Assay ◽

Nucleocapsid Proteins

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Detection of specific IgG, IgM, IgE and IgG subclass antibodies for serological diagnosis of human cystic echinococcosis

Helminthologia ◽

10.1515/helmin-2015-0016 ◽

2015 ◽

Vol 52 (2) ◽

pp. 85-88 ◽

Cited By ~ 1

Author(s):

M. I. Naik ◽

R. Kumar Tenguria ◽

Ehtishamul Haq

Keyword(s):

Cystic Echinococcosis ◽

Diagnostic Sensitivity ◽

Antibody Detection ◽

Enzyme Linked Immunosorbent Assay ◽

Parasitic Infections ◽

Igg Subclass ◽

Healthy Controls ◽

Serum Samples ◽

Igg Antibody ◽

Specific Antibodies

SummaryAn enzyme linked immunosorbent assay (ELISA), based on sheep hydatid cyst fluid antigen was used for the detection of specific antibodies of IgG, IgM, IgE and IgG subclass in the serum samples of 62 clinically and radiologically diagnosed cystic echinococcosis (CE) patients, 8 clinically suspected cases of CE, 25 other parasitic disease controls and 25 healthy controls. The diagnostic sensitivity in the clinically and radiologically suggestive cases (n = 62) for IgG antibody detection was highest (93 %), followed by IgE, IgG4, IgG1, IgG2, IgM and IgG3 with 89 %, 87 %, 85 %, 76 %, 70 % and 55 % respectively. The detection of specific IgE, IgG1 and IgG4 antibody showed the higher diagnostic sensitivity and specificity to the extent that they can be safely used as better substitute to IgG. Even though, the diagnostic sensitivity of IgG was highest (93%) but was less specific (88 %) due to the frequent non-specific reactions in the sera of patients with other parasitic infections and healthy controls. None of the sera samples from healthy controls gave non-specific reaction with IgE, IgG1 and IgG4 and there was a considerably reduced cross-reaction with these antibodies. The most discriminatory and specific antibodies found in this study belonged to IgE, IgG1 and IgG4; therefore, these antibodies may serve as useful diagnostic markers for CE.

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Evaluation of Immunoglobulin G Subclass Antibodies against Recombinant Fasciola gigantica Cathepsin L1 in an Enzyme-Linked Immunosorbent Assay for Serodiagnosis of Human Fasciolosis

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.12.10.1152-1156.2005 ◽

2005 ◽

Vol 12 (10) ◽

pp. 1152-1156 ◽

Cited By ~ 17

Author(s):

Chaisiri Wongkham ◽

Chairat Tantrawatpan ◽

Pewpan M. Intapan ◽

Wanchai Maleewong ◽

Sopit Wongkham ◽

...

Keyword(s):

Immunosorbent Assay ◽

Immunoglobulin G ◽

Fasciola Gigantica ◽

Enzyme Linked Immunosorbent Assay ◽

Parasitic Infections ◽

Predictive Values ◽

Diagnostic Potential ◽

Diagnostic Values ◽

Sensitivity Specificity ◽

Igg4 Antibody

ABSTRACT A cystatin capture enzyme-linked immunosorbent assay (ELISA) using recombinant Fasciola gigantica cathepsin L1 antigen was developed to detect specific immunoglobulin G (IgG) subclass antibodies (IgG1, IgG2, IgG3, and IgG4) and was evaluated for its diagnostic potential for human fasciolosis. In an analysis of the sera of 13 patients infected with F. gigantica, 209 patients with other parasitic infections, 32 cholangiocarcinoma patients, and 42 healthy controls, the IgG4-ELISA gave the highest diagnostic values. The sensitivity, specificity, accuracy, and positive and negative predictive values of this method based on the detection of IgG4 antibody were 100%, 99.3%, 99.3%, 86.7%, and 100%, respectively. The results revealed that restricting the ELISA to the detection of specific IgG4 antibody enhanced the specificity and accuracy for the serodiagnosis of human fasciolosis.

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Enzyme-linked immunosorbent assay determination of specific rubella antibody levels in micrograms of immunoglobulin G per milliliter of serum in clinical samples

Journal of Clinical Microbiology ◽

10.1128/jcm.8.4.419-423.1978 ◽

1978 ◽

Vol 8 (4) ◽

pp. 419-423

Author(s):

P O Leinikki ◽

I Shekarchi ◽

P Dorsett ◽

J L Sever

Keyword(s):

Immunosorbent Assay ◽

Immunoglobulin G ◽

Solid Phase ◽

Laboratory Diagnosis ◽

Clinical Samples ◽

Enzyme Linked Immunosorbent Assay ◽

Igg Antibody ◽

Standard Curve ◽

Antibody Levels

A "microgram assay" is described in which solid-phase enzyme-linked immunosorbent assay is used for the determination of specific rubella immunoglobulin G (IgG) antibody levels in micrograms per milliliter of serum. The quantitation was based on a standard curve obtained by using a reference serum, for which the specific IgG content was assayed by immunochemical purification. IgG was first purified and specific rubella antibodies were separated by an immunoadsorbent prepared by linking rubella virus antigens to Sepharose 4B. By using IgG-specific conjugate, the levels of specific rubella IgG antibodies could then be determined from clinical samples. Seronegative samples showed antibody levels less than 1 microgram/ml, whereas levels up to several hundred micrograms per milliliter were detected in some postinfection sera. The correlation between microgram antibody levels and hemagglutination inhibition titers was linear. The method offers a simple and sensitive antibody assay which could be used both for the laboratory diagnosis of acute rubella and for the evaluation of immunity.

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Detection of Antibodies to BrucellaCytoplasmic Proteins in the Cerebrospinal Fluid of Patients with Neurobrucellosis

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.6.5.756-759.1999 ◽

1999 ◽

Vol 6 (5) ◽

pp. 756-759 ◽

Cited By ~ 29

Author(s):

Pablo C. Baldi ◽

George F. Araj ◽

Graciela C. Racaro ◽

Jorge C. Wallach ◽

Carlos A. Fossati

Keyword(s):

Cerebrospinal Fluid ◽

Western Blotting ◽

Immunosorbent Assay ◽

Immunoglobulin G ◽

Serological Diagnosis ◽

Neurological Involvement ◽

Enzyme Linked Immunosorbent Assay ◽

Specific Diagnosis ◽

Serum Igg ◽

Cytoplasmic Proteins

ABSTRACT The diagnosis of human neurobrucellosis usually relies on the detection of antibodies to Brucella lipopolysaccharide (LPS) in cerebrospinal fluid (CSF) by agglutination tests or enzyme-linked immunosorbent assay (ELISA). Here we describe the detection of immunoglobulin G (IgG) to cytoplasmic proteins (CP) ofBrucella spp. by ELISA and Western blotting in seven CSF samples from five patients with neurobrucellosis. While IgG to CP (titers of 200 to 12,800) and IgG to LPS (800 to 6,400) were found in the CSF of these patients, these antibodies were not detected in CSF samples from two patients who had systemic brucellosis without neurological involvement. The latter, however, had serum IgG and IgM to both LPS and CP. No reactivity to these antigens was found in CSF samples from 14 and 20 patients suffering from nonbrucellar meningitis and noninfectious diseases, respectively. These findings suggest that, in addition to its usefulness in the serological diagnosis of human systemic brucellosis, the ELISA with CP antigen can be used for the specific diagnosis of human neurobrucellosis.

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Changes in Avidity and Level of Immunoglobulin G Antibodies to Mycobacterium tuberculosis in Sera of Patients Undergoing Treatment for Pulmonary Tuberculosis

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.10.4.702-709.2003 ◽

2003 ◽

Vol 10 (4) ◽

pp. 702-709 ◽

Cited By ~ 18

Author(s):

Lenka M. Pereira Arias-Bouda ◽

Sjoukje Kuijper ◽

Anouk Van Der Werf ◽

Lan N. Nguyen ◽

Henk M. Jansen ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Pulmonary Tuberculosis ◽

Immunoglobulin G ◽

Affinity Maturation ◽

Enzyme Linked Immunosorbent Assay ◽

Duration Of Symptoms ◽

Antibody Avidity ◽

Tuberculosis Patients ◽

Specific Antibodies ◽

Antibody Levels

ABSTRACT Much is known about specific antibodies and their titers in patients with tuberculosis. However, little is known about the avidity of these antibodies or whether changes in avidity occur during the progression of the disease or during treatment. The aims of this study were to determine the avidity of antibodies to Mycobacterium tuberculosis in patients with pulmonary tuberculosis, to explore the value of avidity determination for the diagnosis of tuberculosis, and to study changes in levels of antibodies and their avidity during treatment. Antibody avidity was measured by an enzyme-linked immunosorbent assay with thiocyanate elution. Avidity indices and serum levels of immunoglobulin G to M. tuberculosis were determined for 22 patients with pulmonary tuberculosis before and during treatment and for 24 patients with other pulmonary diseases. Antibody levels and avidity were both significantly higher in untreated tuberculosis patients than in the controls. Avidity determination had more diagnostic potential than determination of the antibody levels. Tuberculosis patients with a long duration of symptoms had higher antibody avidity than those with a recent onset of symptoms, indicating affinity maturation of specific antibodies during active disease. In the early phase of treatment, a decrease in antibody avidity was observed for 73% of all tuberculosis patients, accompanied by an initial increase in antibody levels in 36% of these patients. These phenomena could be explained by an intense stimulation of the humoral response by antigens released from killed bacteria, reflecting early bactericidal activity of antituberculous drugs leading to the production of low-affinity antibodies against these released antigens.

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