The role of a recombinant hybrid protein based ELISA for the serodiagnosis of<i>Onchocerca volvulus</i>

Andrews, J A; Bligh, WJ; Chiodini, Peter L.; Bradley, Janette E.; Nde, Pius N.; Lucius, Richard

doi:10.1136/jcp.2006.037093

Cited by 10 publications

(8 citation statements)

References 13 publications

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“…In general, increasing the number of antigens within a mixture is generally a favorable condition if each member antigen has a comparable low background signal. In contrast, ELISAs perform poorly when more than one antigen is immobilized and the alternative is to employ complicated multi-epitope hybrid molecules from different Ov antigens [14],[15]. Luminex technology can also be used in screening panels of antigens for diagnosis, but require much more expensive equipment and thus has little value for detection of onchocerciasis in most non-hospital or research settings.…”

Section: Discussionmentioning

confidence: 99%

“…For example, a field-applicable diagnostic card immunoassay based on a recombinant antigen, Ov-16, showed 80% sensitivity for detecting Ov -infected sera [12],[13], while an ELISA employing a recombinant hybrid Ov protein showed 93% sensitivity [14],[15]. In these and other studies, issues related to cross-reactivity with other filarial-infected sera such as with Wb and Ll were not studied with a large enough group of samples and generally showed poorer discrimination of Ov -infected sera from these other infections.…”

Section: Discussionmentioning

confidence: 99%

“…Each antigen, when tested, has had the characteristic of identifying Ov infection early (often pre-patency) in the infection. More recently a field-applicable diagnostic immunoassay based on one of these Ov-specific recombinant antigens, Ov-16, showed 80% sensitivity for detecting Ov -infected sera [12],[13], while an ELISA employing a recombinant hybrid Ov protein showed 93% sensitivity [14],[15]. Despite the high sensitivity of all these immunoassays, each of these tests have had some difficulty discriminating Ov -infected sera from some other filarial infections that can be co-endemic with O. volvulus such as Wuchereria bancrofti (a causal agent of lymphatic filariasis) and L. loa (the causal agent of loiasis).…”

Section: Introductionmentioning

confidence: 99%

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A Four-Antigen Mixture for Rapid Assessment of Onchocerca volvulus Infection

et al. 2009

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BackgroundOnchocerciasis, an infection caused by the filarial nematode Onchocerca volvulus, is a major public health concern. Given the debilitating symptoms associated with onchocerciasis and concerns about recrudescence in areas of previous onchocerciasis control, more efficient tools are needed for diagnosis and monitoring of control measures. We investigated whether luciferase immunoprecipitation systems (LIPS) may be used as a more rapid, specific, and standardized diagnostic assay for Onchocerca volvulus infection.MethodsFour recombinantly produced Onchocerca volvulus antigens (Ov-FAR-1, Ov-API-1, Ov-MSA-1 and Ov-CPI-1) were tested by LIPS on a large cohort of blinded sera comprised of both uninfected controls and patients with a proven parasitic infection including Onchocerca volvulus (Ov), Wuchereria bancrofti (Wb), Loa loa (Ll), Strongyloides stercoralis (Ss), and with other potentially cross-reactive infections. In addition to testing all four Ov antigens separately, a mixture that tested all four antigens simultaneously was evaluated in the standard 2-hour incubation format as well as in a 15-minute rapid LIPS format.FindingsAntibody responses to the four different Ov antigens allowed for unequivocal differentiation between Ov-infected and uninfected control sera with 100% sensitivity and 100% specificity. Analysis of the antibody titers to each of these four antigens in individual Ov-infected sera revealed that they were markedly different and did not correlate (rS = –0.11 to 0.58; P = 0.001 to 0.89) to each other. Compared to Ov-infected sera, patients infected with Wb, Ll, Ss, and other conditions had markedly lower geometric mean antibody titers to each of the Ov 4 antigens (P<0.0002 for each antigen). The simplified method of using a mixture of the 4 Ov antigens simultaneously in the standard format or a quick 15-minute format (QLIPS) showed 100% sensitivity and 100% specificity in distinguishing the Ov-infected sera from the uninfected control sera. Finally, the QLIPS format had the best performance with 100% sensitivity and specificity values of 76%, 84% and 93% for distinguishing Ov from Wb, Ll and Ss-infected sera.ConclusionsThe multi-antigen LIPS assay can be used as a rapid, high throughput, and specific tool to not only to diagnose individual Ov infections but also as a sensitive and potentially point-of-care method for early detection of recrudescent infections in areas under control and for mapping new areas of transmission of Ov infection.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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A Four-Antigen Mixture for Rapid Assessment of Onchocerca volvulus Infection

et al. 2009

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“…Skin snips placed in saline to detect larva, examine surgically removed nodules for adult worms ELISA 90,91 Nested PCR 112 Taenia solium Detection of eggs in stool ELISA, 92,93,96 immunoblot [94][95][96] Nested PCR 113 MS 123 marrow, or lymph nodes. 15 Using spleen samples increases sensitivity, but the procedure to obtain the aspirates risks internal bleeding.…”

Section: Onchocerca Volvulusmentioning

confidence: 99%

Diagnosis of Parasitic Infections: What’s Going On?

Ricciardi

Ndao

2015

SLAS Discovery

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Methods for the diagnosis of parasitic infections have stagnated in the past three decades. Labor-intensive methods such as microscopy still remain the mainstay of several diagnostic laboratories. There is a need for more rapid tests that do not sacrifice sensitivity and that can be used in both clinical settings as well as in poor resource field settings. The fields of diagnostic medical parasitology, treatment, and vaccines are undergoing dramatic change. In recent years, there has been tremendous effort to focus research on the development of newer diagnostic methods focusing on serological, molecular, and proteomic approaches. This article examines the various diagnostic tools that are being used in clinical laboratories, optimized in reference laboratories, and employed in mass screening programs.

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“…Efforts to simplify testing in an ELISA format by coating multiple antigens to the microtiter plates are usually unsuccessful because the antigen coating efficiency is quite variable, resulting in poor diagnostic performance. Alternatively, multiepitope hybrid recombinant molecules have been employed, whereby multiple antigens or immunodominant regions are fused together in a single protein [47–49]. While these hybrid antigens can be highly useful, time and effort is required to design them and optimize their performance.…”

Section: Lips Antibody Profilingmentioning

confidence: 99%

Antibody-profiling technologies for studying humoral responses to infectious agents

Burbelo

Ching

Bush

et al. 2010

Expert Review of Vaccines

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Analyses of humoral responses against different infectious agents are critical for infectious disease diagnostics, understanding pathogenic mechanisms, and the development and monitoring of vaccines. While ELISAs are often used to measure antibody responses to one or several targets, new antibody-profiling technologies, such as protein microarrays, can now evaluate antibody responses to hundreds, or even thousands, of recombinant antigens at one time. These large-scale studies have uncovered new antigenic targets, provided new insights into vaccine research and yielded an overview of immunoreactivity against almost the entire proteome of certain pathogens. However, solid-phase antigen arrays also have drawbacks that limit the type of information obtained, including suboptimal detection of conformational epitopes, high backgrounds due to impure antigens and a narrow dynamic range of detection. We have developed a solution-phase antibody-profiling technology, luciferase immunoprecipitation systems (LIPS), which harnesses light-emitting recombinant antigen fusion proteins to quantitatively measure patient antibody titers. Owing to the highly linear light output of the luciferase reporter, some antibodies can be detected without serum dilution in a dynamic range of detection often spanning seven orders of magnitude. When LIPS is applied iteratively with multiple target antigens, a high-definition antibody profile is obtained. Here, we discuss the application of these different antibody-profiling technologies and their associated limitations with particular emphasis on protein microarrays. We also describe LIPS in detail and discuss several clinically relevant uses of the technology. Together, these new technologies offer new tools for understanding humoral responses to known and emerging infectious agents.

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The role of a recombinant hybrid protein based ELISA for the serodiagnosis ofOnchocerca volvulus

Abstract: This study demonstrates the potential role of the assay as a sensitive and specific test for use in the serodiagnosis of onchocerciasis in a reference laboratory dealing with sera from patients in non-endemic setting.

Cited by 10 publications

References 13 publications

A Four-Antigen Mixture for Rapid Assessment of Onchocerca volvulus Infection

A Four-Antigen Mixture for Rapid Assessment of Onchocerca volvulus Infection

Diagnosis of Parasitic Infections: What’s Going On?

Antibody-profiling technologies for studying humoral responses to infectious agents

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