Development of a robust and standardized immunoserological assay for detection of anti-measles IgG antibodies in human sera

开发一种强大且标准化的免疫血清学检测方法,用于检测人类血清中的抗麻疹 IgG 抗体

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作者:Katalin Böröcz, Zsuzsanna Csizmadia, Ákos Markovics, Viktória Mészáros, Kornélia Farkas, Vivien Telek, Vivien Varga, Geofrey Ouma Maloba, Kornélia Bodó, József Najbauer, Tímea Berki, Péter Németh

Abstract

Because of measles outbreaks there is a need for continuous monitoring of immunological protection against infection at population level. For such monitoring to be feasible, a cost-effective, reliable and high-throughput assay is necessary. Herein we describe an ELISA protocol for assessment of anti-measles antibody levels in human serum samples that fulfills the above criteria and is easily adaptable by various laboratories. A serum bank of anonymous patient sera was established (N > 3000 samples). Sera were grouped based on measles immunization schedules and/or changes in vaccine components since the introduction of the first measles vaccine in Hungary in 1969. Newly designed ELISA was performed by using Siemens BEP 2000 Advance System and data were confirmed using commercially available kits. Our indirect ELISA was compared to indirect immunfluoresence and to anti-measles nucleocapsid (N) monoclonal antibody-based sandwich ELISA. The results obtained are in high agreement with the confirmatory methods, and reflect measles vaccination history in Hungary ranging from pre-vaccination era, through the initial period of measles vaccination, to present. Based on measurement of 1985 sera, the highest ratio of low/questionable antibody level samples was detected in cluster '1978-1987' (~25.4%), followed by cluster '1969-1977' (~15.4%).Our assay is suitable for assessment of anti-measles immunity in a large cohort of subjects. The assay is cost-effective, allows high-throughput screening and has superior signal-to-noise ratio. This assay can serve as a first step in assessment of the effectiveness of all three components of the MMR vaccine.

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