Escherichia coli O157 serology: false-positive ELISA results caused by human antibodies binding to bovine serum albumin

Chart, Henrik; Evans, J.; Chalmers, Rachel M.; Salmon, R. L.

doi:10.1046/j.1472-765x.1998.00396.x

Cited by 10 publications

(8 citation statements)

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“…For example, in a study that was examining for the presence of human antibodies to Japanese encephalitis virus, antibodies that cross-reacted with BSA were found (Konishi et al, 2010). Another study found that human antibodies non-specifically bound to BSA (A2153 (≥ 96% pure preparation from Sigma)) (Chart et al, 1998). To prevent falsepositive results from either cross reactive antibodies or from non-specific binding of ELISA reagents to BSA, alternative blocking agents can be used.…”

Section: Discussionmentioning

confidence: 99%

“…To prevent falsepositive results from either cross reactive antibodies or from non-specific binding of ELISA reagents to BSA, alternative blocking agents can be used. Examples include rabbit serum (Chart et al, 1998), casein (Konishi et al, 2010), heat-denaturing the blocking protein (Mauracher et al, 1991), non-protein blocking solutions like Synblock (Afrough et al, 2007) or Ficoll or polyvinylalcohol (Huber et al, 2009). Alternatively, no protein can be included in the blocking buffer (Jorgensen et al, 2005), although this too can potentially cause artifactual results (Bird et al, 1988).…”

Section: Discussionmentioning

confidence: 99%

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Enzyme-linked immunosorbent assay (ELISA) and blocking with bovine serum albumin (BSA)—not all BSAs are alike

Xiao

Isaacs

2012

Journal of Immunological Methods

147

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The enzyme-linked immunosorbent assays (ELISA) is an extremely common and powerful laboratory technique for detecting proteins by antibodies. Researchers frequently use bovine serum albumin (BSA) as a blocking agent to prevent non-specific binding of antigens and antibodies to the microtiter well. While studying the interactions of the vaccinia virus complement control protein (VCP) with complement, we found nonspecific binding of VCP to BSA and identify a BSA preparation that did not result in non-specific binding. This work draws attention to the fact that not all BSA preparations are alike. It also highlights the need to perform critical controls to ensure that ELISA reactants do not inappropriately bind to the blocking agent.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Enzyme-linked immunosorbent assay (ELISA) and blocking with bovine serum albumin (BSA)—not all BSAs are alike

Xiao

Isaacs

2012

Journal of Immunological Methods

147

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“…were Cryptosporidium parvum (previously referred to as C. interfere with the test (Venkatesan & Wakelin 1993;Chart et al 1998). The studies by and Mead et al (1988) used western blotting that enabled identification of important antigen molecules.…”

Section: Discussion Of Early Worldwide Studiesmentioning

confidence: 99%

“…A note of caution, however, is that the method used requires blocking non-specific binding with BSA, to which dairy farmers may have antibodies (Chart et al 1998). …”

Section: Early Worldwide Studiesmentioning

confidence: 99%

Towards a US national estimate of the risk of endemic waterborne disease – sero-epidemiologic studies

Casemore

2006

Journal of Water and Health

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Worldwide literature on serological methods and sero-surveys on waterborne pathogens has been reviewed. Outbreak investigation and research reports have also been examined to aid understanding of the serological response and transmission dynamics. The aim was to seek an estimate of seroprevalence and to determine if this could inform the US national estimate of risk for endemic waterborne infection associated with public water supplies. Antibody responses indicate infection, both symptomatic and asymptomatic, so probably give a truer indication of prevalence. Outbreak data can probably be regarded as the upper bound for seroprevalence estimations. Antibody is not necessarily protective per se but is a good indicator for at least partial resistance to symptomatic infection; absence of antibody will normally imply susceptibility.Pathogens transmitted by water are commonly transmitted by other routes. However, the fact that other transmission routes are more common does not detract from the potential protective effect of immunity when waterborne transmission occurs. Data indicate that seroprevalence varies widely, reflecting geographic, social and hygiene factors, but is generally greater where surface water sources are used rather than groundwater. Areas of low seroprevalence may expect a high attack rate in the event of contamination of their water supply.Key words | Campylobacter, Cryptosporidium, ELISA, endemic infection, E. coli 0157, Giardia, molecular studies, noroviruses, outbreaks, sero-epidemiology, seroprevalence, waterborne pathogens, western blot ABBREVIATIONS DEFINITIONS AND GENERAL ASPECTS OF IMMUNOLOGYEpidemiology, at its simplest, is the study of the occurrence (number and frequency) and distribution of cases by person This paper is in the public domain: verbatim copying and redistribution of this paper are permitted in all media for any purpose, provided this notice is preserved along with the paper's original DOI. Anyone using the paper is requested to properly cite and acknowledge the source as J. The outcome of the contamination of water supply and transmission to consumers depends on complex dynamics with various, usually ill characterised, factors-the organism, the exposed persons (and their levels of immunity) and the environment (Casemore 1994(Casemore , 1995Meinhardt et al. 1996).In order to better understand the natural history of waterborne infections and the complexities of the dynamics of transmission (see Figure 1), epidemiological investigations can be enhanced by serologic studies . The purpose of this paper concerns the national estimate of risk of endemic infection from water. However, reports on immunological studies and on the investigation of outbreaks are fundamental to the understanding of endemic infection and illness. In general terms, prevalence in outbreaks likely represents the upper boundary for endemic infection estimates.Sero-epidemiology is a form of study in which the demonstration of antibodies to the organism of interest is used as a surrogate or marker for the d...

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“…The simplest explanation may be insufficient purity of an antigen used for microplate coating, or too low initial sample dilution before the test (especially in the case of relatively low cut-off value). Some authors suggested that non-specific binding of serum antibodies with some blocking agents used in ELISA production may occur (Chart et al, 1998;Konishi et al, 2010). The reason for the occurrence of false-positive results in non-competitive ELISAs may also be attributed to a phenomenon known as a bridging by heterophilic antibodies (e.g.…”

Section: Discussionmentioning

confidence: 99%

Comparison of six commercial ELISAs for the detection of antibodies against porcine reproductive and respiratory syndrome virus (PRRSV) in field serum samples

Biernacka

Podgórska

Tyszka³

et al. 2018

Research in Veterinary Science

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Porcine reproductive and respiratory syndrome (PRRS) is one of the most common infectious diseases of swine globally. Since the course of PRRS virus (PRRSV) infection is subclinical, laboratory diagnosis is necessary to detect the virus or specific antibodies. The aim of this study was to assess the sensitivity and specificity of IDEXX PRRS X3 Ab Test (IDEXX, USA), Civtest Suis E/S (Hipra, Spain), INgezim PRRS 2.0 (Ingenasa, Spain), VetExpert PRRS Ab ELISA 4.0 (BioNote, Korea), Pigtype PRRSV Ab (Qiagen, Germany) and PrioCHECK PRRSV Antibody ELISA (ThermoFisher, USA), using serum samples obtained from 5 conventional PRRSV-positive and 5 PRRSVnegative Polish pig farms. Specificity of ELISAs ranged from 94.2% (ThermoFisher) to 100% (IDEXX and Hipra). ThermoFisher ELISA had the highest detection rate and detected 67.2% samples from PRRSV-positive farms as positive but considering its low specificity some of the positive results may be incorrect. IDEXX ELISA considered as a reference detected 64.8% positive sera in PRRSV-positive farms. On the other hand Hipra Elisa identified only 51.8% of samples as positive. The diagnostic sensitivity of five ELISAs relative to IDEXX ranged from 80.3% (Hipra) to 96.3% (ThermoFisher). Our study showed significant differences in specificity and diagnostic sensitivity between the compared kits. The differences in the performance appeared to be practically negligible on farms where early infection with PRRSV occurred. However, on PRRSV-negative farms, or farms with PRRSV stable sow herds, some ELISAs can give results not reflecting the infection status in specific age groups.

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Escherichia coli O157 serology: false-positive ELISA results caused by human antibodies binding to bovine serum albumin

Cited by 10 publications

References 11 publications

Enzyme-linked immunosorbent assay (ELISA) and blocking with bovine serum albumin (BSA)—not all BSAs are alike

Enzyme-linked immunosorbent assay (ELISA) and blocking with bovine serum albumin (BSA)—not all BSAs are alike

Towards a US national estimate of the risk of endemic waterborne disease – sero-epidemiologic studies

Comparison of six commercial ELISAs for the detection of antibodies against porcine reproductive and respiratory syndrome virus (PRRSV) in field serum samples

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