A recombinant antigen-based IgG4 ELISA for the specific and sensitive detection of Brugia malayi infection

Rahmah, N.; Lim, Boon Huat; Anuar, A. Khairul; Shenoy, R. K.; Kumaraswami, V.; Hakim, S Lokman; Chotechuang, P.; Kanjanopas, Kobkan; Ramachandran, C. P.

doi:10.1016/s0035-9203(01)90234-2

Cited by 45 publications

(50 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Some of these immunologically relevant proteins have been described previously (23-33) and relate to those molecules that are highly immunogenic in human infections or that have activity on the mammalian host immune response (through mimicry or other mechanisms). These data corroborate a number of previous studies that demonstrate stage-specific expression and/or serologic reactivity of ALT-2 and the larval allergens (in the L3 stage) (34) and BmR1 (35), BmMIF (36), TGF-β homologue (30), SXP-1 (37), galectins (38), and microfilarial sheath proteins (in the microfilarial stage) (39). Particular sets of ECM-associated proteins were common and highly enriched in the both the L3s (SI Appendix, Fig.…”

Section: Resultssupporting

confidence: 91%

Stage-specific proteomic expression patterns of the human filarial parasiteBrugia malayiand its endosymbiontWolbachia

Bennuru

Meng

Ribeiro

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

123

View full text Add to dashboard Cite

Global proteomic analyses of pathogens have thus far been limited to unicellular organisms (e.g., protozoa and bacteria). Proteomic analyses of most eukaryotic pathogens (e.g., helminths) have been restricted to specific organs, specific stages, or secretomes. We report here a large-scale proteomic characterization of almost all the major mammalian stages of Brugia malayi, a causative agent of lymphatic filariasis, resulting in the identification of more than 62% of the products predicted from the Bm draft genome. The analysis also yielded much of the proteome of Wolbachia, the obligate endosymbiont of Bm that also expressed proteins in a stage-specific manner. Of the 11,610 predicted Bm gene products, 7,103 were definitively identified from adult male, adult female, blood-borne and uterine microfilariae, and infective L3 larvae. Among the 4,956 gene products (42.5%) inferred from the genome as "hypothetical," the present study was able to confirm 2,336 (47.1%) as bona fide proteins. Analysis of protein families and domains coupled with stage-specific expression highlight the important pathways that benefit the parasite during its development in the host. Gene set enrichment analysis identified extracellular matrix proteins and those with immunologic effects as enriched in the microfilarial and L3 stages. Parasite sex-and stage-specific protein expression identified those pathways related to parasite differentiation and demonstrates stage-specific expression by the Bm endosymbiont Wolbachia as well.filaria | nematode D isease associated with infection by Brugia malayi (Bm) and Wuchereria bancrofti, the two major causative organisms of human lymphatic filariasis, is the second leading cause of morbidity/disability worldwide, in large part because of the parasites' ability to alter the structural and functional integrity of the lymphatics, leading to lymphedema and elephantiasis. Invasion, establishment of infection within the host and development are essential processes within the complex parasite life cycle (SI Appendix, Fig. S1), with many of the parasitic stages being targets for therapeutic intervention or vaccines. Each of the filarial life cycle stages has characteristics that are shared and others that are stage-specific.Filarial infections are often characterized by a series of discrete host responses directed at the parasite and its endosymbiont Wolbachia that evolve during the course of infection. Because proteins are usually the effectors of most biological functions, proteomic data enable a more direct understanding of these important processes compared with those inferred from genomic studies. Absolute quantification of genome-wide expressed proteins is not yet within our reach for most eukaryotes. However, spectral counts of massive MS-based data (e.g., observed frequencies of each peptide) allow for relative quantification. Proteomic data also allow for clearer genomic curation by improving annotation and the identification of translational sites, stop codon read-throughs, frame shifts, and predict...

show abstract

Section: Resultssupporting

confidence: 91%

Stage-specific proteomic expression patterns of the human filarial parasiteBrugia malayiand its endosymbiontWolbachia

Bennuru

Meng

Ribeiro

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

123

View full text Add to dashboard Cite

show abstract

“…The IgG assay, however, provides false-positive reactions with other parasitic helminths, whereas the IgG4 antibody greatly increased the specificity of the assay for toxocariasis (15,16,26). Similarly, in the diagnosis of lymphatic filariasis, antifilarial IgG4 is often used as a marker of active infection (8,9,19,25), but to date, there is no commercially available IgG4 test for the detection of Toxocara infection.…”

Section: Discussionmentioning

confidence: 99%

Development and Evaluation of a Sensitive and Specific Assay for Diagnosis of Human Toxocariasis by Use of Three Recombinant Antigens (TES-26, TES-30USM, and TES-120)

2009

View full text Add to dashboard Cite

Diagnosis of human toxocariasis currently relies on serologic tests that use Toxocara excretory-secretory (TES) antigen to detect immunoglobulin G (IgG) antibodies to the larvae. In general, however, these assays do not have adequate specificity for use in countries in which other soil-transmitted helminths are endemic. The use of recombinant antigens in these assays, however, is promising for improving the specificity of the diagnosis of toxocariasis. Toward this goal, we developed an IgG4 enzyme-linked immunosorbent assay (ELISA) involving three recombinant antigens: rTES-30USM (previously produced), rTES-26, and rTES-120. The latter two antigens were produced by reverse transcription-PCR cloning; subcloned into glutathione S-transferase (GST)-tagged and His-tagged prokaryotic expression vectors, respectively; and expressed in Escherichia coli. The recombinant proteins were subsequently purified by affinity chromatography using GST and His-Trap resins. The diagnostic potential of each purified recombinant antigen was tested with various immunoglobulin classes (IgG, IgM, and IgE) and IgG subclasses. The IgG4 ELISA was determined to have the highest specificity and was further evaluated using a panel of serum samples. The rTES-26 IgG4 ELISA showed 80.0% (24/30 samples positive) sensitivity, and both the rTES-30USM IgG4 ELISA and rTES-120 IgG4 ELISA had 93.0% (28/30) sensitivity. Combined use of rTES-120 and rTES-30 IgG4 ELISA for the diagnosis of toxocariasis provided 100% sensitivity. The specificities of rTES-26, rTES-30USM, and rTES-120 antigens were 96.2%, 93.9%, and 92.0%, respectively. These results indicate that the development of a diagnostic test using the three recombinant antigens will allow for more-accurate detection of toxocariasis.Human toxocariasis is a worldwide parasitic zoonosis, caused most commonly by the intestinal parasites the dog roundworm (Toxocara canis) and also the cat roundworm (Toxocara cati) (2). It commonly manifests as visceral larva migrans, ocular larva migrans, and covert toxocariasis. Toxocariasis is probably one of the most common zoonotic helminthiases in temperate climates and developed countries (20). In Malaysia, the seroprevalence rate is approximately 20% (6).Definitive diagnosis of toxocariasis is based on the detection of Toxocara larvae from biopsy tissues, but this test is timeconsuming and difficult to perform. Therefore, diagnosis is commonly based on clinical and serologic diagnosis. Currently, common routine serology tests are applied for detection, such as commercial immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) kits in which Toxocara excretorysecretory (TES) antigens obtained from culture of T. canis second-stage (L2) larvae are used (22). The use of the native TES antigen for serodiagnosis of human toxocariasis is a laborious and time-consuming technique, and the production capacity is limited by the culture volume (22). Further, the specificity is often low, especially in developing countries, where infections with helminths that cause c...

show abstract

“…having various degrees of sensitivity and specificity have been reported (5,10,11,18,19,22,25). Attempts have been made to develop immunodiagnostic test kits for the detection of circulating filarial antigen for the identification of active filarial infection.…”

mentioning

confidence: 99%

“…However these commercial reagents have the limitation of being able to detect only bancroftian infection. More recently a recombinant antigen-based rapid IgG4 antibody dipstick test has been developed for the detection of antibodies in sera from patients with brugian infections (18,19). This test showed only 54.5% positive reaction with individuals having W. bancrofti infection.…”

mentioning

confidence: 99%

Development and Evaluation of a Rapid Flow‐Through Immuno Filtration Test Using Recombinant Filarial Antigen for Diagnosis of Brugian and Bancroftian Filariasis

Baskar

Srikanth

Suba

et al. 2004

Microbiology and Immunology

View full text Add to dashboard Cite

There is an imperative need to develop a rapid antibody test that can be used for diagnosis of clinical cases in travelers and expatriates, primary surveillance in areas of unknown endemicity, detection of early infection in childhood and for monitoring chemotherapeutic programs. A rapid‐format, simple and qualitative flow through immuno filtration test has been developed for the identification of total IgG antibodies to recombinant filarial antigen WbSXP‐1. This test system employs colloidal gold‐protein A reagent as the antibody capture reagent. The sensitivity and specificity of the test was evaluated in a total of 1,230 serum samples. The sensitivity of the test was found to be 90.8% with brugian (n=70) and 91.4% with bancroftian (n = 140) microfilaraemic subjects. The test showed minimum reactivity (4/10) with Loa loa microfilaria (MF) positive sera and no reactivity (0/20) with Onchocerca MF positive sera. This rapid diagnosis is found to be non‐reactive with individuals having other parasitic diseases including schistosomiasis (n = 10), soil‐transmitted helminthiases (n=34) and protozoan infections (n=33) indicating the potential of this test as a prospective method of diagnosis for both brugian and bancroftian lymphatic filariasis. Stability kinetics was studied at different temperatures and different time periods. The rapid flow‐through immuno filtration test is advantageous since it can be stored at room temperature, is user friendly and is particularly applicable in the field as an initial screening method, for epidemiological monitoring of filarial infections in bancroftian and brugian endemic regions of the world.

show abstract

A recombinant antigen-based IgG4 ELISA for the specific and sensitive detection of Brugia malayi infection

Cited by 45 publications

References 16 publications

Stage-specific proteomic expression patterns of the human filarial parasiteBrugia malayiand its endosymbiontWolbachia

Stage-specific proteomic expression patterns of the human filarial parasiteBrugia malayiand its endosymbiontWolbachia

Development and Evaluation of a Sensitive and Specific Assay for Diagnosis of Human Toxocariasis by Use of Three Recombinant Antigens (TES-26, TES-30USM, and TES-120)

Development and Evaluation of a Rapid Flow‐Through Immuno Filtration Test Using Recombinant Filarial Antigen for Diagnosis of Brugian and Bancroftian Filariasis

Contact Info

Product

Resources

About