Characterization of Reference Materials for Human Antiserum to Pertussis Antigens by an International Collaborative Study

Xing, Dorothy; König, Carl Heinz Wirsing von; Newland, Penny; Riffelmann, Marion; Meade, Bruce D.; Corbel, Michael J.; Gaines-Das, R.

doi:10.1128/cvi.00372-08

Cited by 60 publications

(55 citation statements)

References 20 publications

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“…Results of quantitative pertussis serology can be expressed in IU/ml (24), and these units correspond to those of reference preparations of CBER/ FDA, which were standardized in ELISA units/ml (EU/ml). Expressing results in IU/ml requires that the kits contain purified antigens and no antigen mixture, such as PT plus FHA.…”

Section: Resultsmentioning

confidence: 99%

“…Studies have focused mostly on antibodies of isotype IgG, and the role of antibodies of isotype IgA especially in a vaccinated population is not totally clear (10,13). Concentrations of antibodies to Bordetella antigens can be measured quantitatively and expressed in international units per ml (IU/ml) according to the 1st International Standard preparation (24).…”

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confidence: 99%

“…Standardization has been performed for in-house ELISAs in two international collaborative studies (12,24). The CDC has recently developed and validated a PT ELISA for use in public health laboratories (14).…”

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confidence: 99%

“…Another 24 samples were from patients with clinical symptoms (more than 2 weeks of coughing) but without laboratory evidence of recent contact with bordetellae (PCR-negative samples, with all antibodies below cutoff values). Eight serum samples were from a vaccine study of adolescents with a Tdap vaccine (10), and one sample contained very low or unmeasurable antibodies to pertussis antigens and was also used in the validation study for the WHO reference preparation (24).…”

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confidence: 99%

“…We chose to compare all ELISA kits that were commercially available and used in Germany in 2008 in relation to the recently available International Reference preparation (24). (ii) Patient sera.…”

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confidence: 99%

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Performance of Commercial Enzyme-Linked Immunosorbent Assays for Detection of Antibodies to Bordetella pertussis

et al. 2010

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Measuring antibodies to Bordetella pertussis antigens is mostly done by enzyme-linked immunosorbent assays (ELISAs). We compared the performance of ELISA kits that were commercially available in Germany. Eleven measured IgG antibodies, and nine measured IgA antibodies. An in-house ELISA with purified antigens served as a reference method. Samples included two WHO reference preparations, the former Food and Drug Administration (FDA)/Center for Biologics Evaluation and Research (CBER) reference preparations, serum samples from patients with clinically suspected pertussis, and serum samples from patients having received a combined tetanus, diphtheria, and pertussis (Tdap) vaccination. Kits using pertussis toxin (PT) as an antigen showed linearity compared to the WHO Reference preparation (r 2 between 0.82 and 0.99), and these kits could quantify antibodies according to the reference preparation. ELISA kits using mixed antigens showed no linear correlation to the reference preparations. Patient results were compared to results of in-house ELISAs using a dual cutoff of either >100 IU/ml anti-PT IgG or >40 IU/ml anti-PT IgG together with >12 IU/ml anti-PT IgA. The sensitivities of kits measuring IgG antibodies ranged between 0.84 and 1.00. The specificities of kits using PT as an antigen were between 0.81 and 0.93. The specificities of kits using mixed antigens were between 0.51 and 0.59 and were thus not acceptable. The sensitivities of kits measuring IgA antibodies ranged between 0.53 and 0.73, and the specificities were between 0.67 and 0.94, indicating that IgA antibodies may be of limited diagnostic value. Our data suggest that ELISAs should use purified PT as an antigen and be standardized to the 1st International Reference preparation.

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Performance of Commercial Enzyme-Linked Immunosorbent Assays for Detection of Antibodies to Bordetella pertussis

et al. 2010

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What to do and what not to do in serological diagnosis of pertussis: recommendations from EU reference laboratories

Guiso

Berbers

Fry

et al. 2010

Eur J Clin Microbiol Infect Dis

210

196

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Bordetella pertussis-specific antibodies can be detected by enzyme-linked immunosorbent assays (ELISAs) or multiplex immunoassays. Assays use purified or mixed antigens, and only pertussis toxin (PT) is specific for B. pertussis. The interpretation of results can be based on dual-sample or single-sample serology using one or two cut-offs. The EU Pertstrain group recommends that: (i) ELISAs and multiplex immunoassays should use purified non-detoxified PT as an antigen, that they should have a broad linear range and that they should express results quantitatively in International Units per millilitre (IU/ml); (ii) a single or dual diagnostic cut-off for single-serum serology using IgG-anti-PT between 50 and 120 IU/ml should be used, and diagnostic serology cannot be validly interpreted for one year after vaccination with acellular pertussis (aP) vaccines; (iii) IgA-anti-PT should only be used with indeterminate IgG-anti-PT levels or when a second sample cannot be obtained. This group discourages using: (i) other antigens in routine diagnostics, as they are not specific; (ii) micro-agglutination, due to its lack of sensitivity; (iii) immunoblots for pertussis serodiagnosis, as results cannot be quantified; (iv) other methods, such as complement fixation or indirect immunofluorescence, due to their low sensitivity and/or specificity.

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Comparison of commercially available immunoblot assays measuring IgG and IgA antibodies to Bordetella pertussis antigens

Kennerknecht

Riffelmann

Schmetz

et al. 2011

Eur J Clin Microbiol Infect Dis

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Bordetella pertussis infection is mostly diagnosed by serological tests, such as by enzyme-linked immunosorbent assays (ELISAs) or by immunoblots. We compared immunoblots from five different manufacturers. Immunoblots from Euroimmun, Mikrogen, Trinity Biotech, Viramed and Virotech were used. All kits except the kit from Trinity Biotech measured IgG and IgA antibodies separately. The kits were used according to the kit inserts. Various reference preparations from the World Health Organization (WHO), the National Institute for Biological Standards and Control (NIBSC) and the Center for Biologics Evaluation and Research/Food and Drug Administration (CBER/FDA) were analysed. Patient sera with high antibody titres in ELISA, sera from patients with compatible clinical symptoms and sera from vaccinees were compared. An algorithm for interpreting quantitative values for IgG and IgA anti-pertussis toxin (PT) from in-house ELISAs was used as a reference. The sensitivity and specificity of the assays was variable when comparing the qualitative results of immunoblots with expected values of reference preparations and ELISA interpretation of patient sera. The interpretation of semi-quantitative reading of the immunoblots did not compare well to the ELISA results. Adenylate cyclase toxin as an additional antigen in two immunoblots did not effectively distinguish between infection and vaccination. Due to the lack of quantification of antibody concentrations, IgG and IgA immunoblots are of limited value in the serological diagnosis of pertussis.

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Characterization of Reference Materials for Human Antiserum to Pertussis Antigens by an International Collaborative Study

Cited by 60 publications

References 20 publications

Performance of Commercial Enzyme-Linked Immunosorbent Assays for Detection of Antibodies to Bordetella pertussis

Performance of Commercial Enzyme-Linked Immunosorbent Assays for Detection of Antibodies to Bordetella pertussis

What to do and what not to do in serological diagnosis of pertussis: recommendations from EU reference laboratories

Comparison of commercially available immunoblot assays measuring IgG and IgA antibodies to Bordetella pertussis antigens

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