metricas
covid
Buscar en
Enfermedades Infecciosas y Microbiología Clínica
Toda la web
Inicio Enfermedades Infecciosas y Microbiología Clínica Comparative evaluation of assays for IgM detection of rubella and measles infect...
Información de la revista
Vol. 40. Núm. 1.
Páginas 22-27 (enero 2021)
Compartir
Compartir
Descargar PDF
Más opciones de artículo
Visitas
3692
Vol. 40. Núm. 1.
Páginas 22-27 (enero 2021)
Original article
Acceso a texto completo
Comparative evaluation of assays for IgM detection of rubella and measles infections
Evaluación comparativa de ensayos para la detección de IgM en infecciones por rubéola y sarampión
Visitas
3692
Mayte Pérez Olmedaa, Pilar Balfagóna, Juan Camachoa, Desirée Dafouza, Jesús de la Fuentea, María Ángeles Murilloa, José Luis Muñoza, Aurora Fernández Garcíab,d, Juan Carlos Sanzc,d, Fernando de Orya,d,
Autor para correspondencia
fernandodeorym@gmail.com

Corresponding author.
a Laboratorio de Serología, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Spain
b Laboratorio de Aislamiento Viral, Centro Nacional de Microbiología, Instituto de Salud Carlos III, Majadahonda, Spain
c Laboratorio Regional de Salud Pública de la Comunidad de Madrid, Madrid, Spain
d Centro de Investigación Biomédica en Red en Epidemiología y Salud Pública (CiberESP), Instituto de Salud Carlos III, Madrid, Spain
Este artículo ha recibido
Información del artículo
Resumen
Texto completo
Bibliografía
Descargar PDF
Estadísticas
Tablas (5)
Table 1. List of assays analyzed.
Table 2. Comparison of rubella assays.
Table 3. Performance characteristics of rubella assays.a
Table 4. Comparison of measles assays.
Table 5. Performance characteristics of measles assaysa.
Mostrar másMostrar menos
Abstract
Background

Serological diagnosis of infections due to measles and rubella viruses is done by IgM detection. The aim of this study was to comparatively evaluate commercial systems for detecting IgM against both viruses, including those of ELISA, in indirect and capture formats, chemiluminescence and electrochemiluminescence.

Methods

Seven (for rubella) and six (for measles) assays were studied. One hundred and sixty two samples were included in the study (from 90 rubella and 72 measles cases), and all were analyzed in all the assays.

Results

The ranges of sensitivity, specificity and agreement for rubella were 94.8–100%, 52.4–100% and 75.5–98.1%, respectively. The corresponding ranges for measles assays were 87.0–100%, 53.3–100%, and 73.0–99.4%.

Conclusion

The best-performing assays were chemiluminescence (for measles and rubella IgM), and electrochemiluminescence (for rubella IgM).

Keywords:
Rubella virus
Measles virus
Enzyme immunoassay
Chemiluminescent immunoassay
Electrochemiluminescent immunoassay
Resumen
Introducción

El diagnóstico serológico de las infecciones por los virus de la rubéola y del sarampión se realiza por detección de IgM específica. El objetivo de este estudio fue evaluar comparativamente sistemas comerciales para la detección de IgM frente a ambos virus, incluyendo ensayos de ELISA, tanto con metodologías indirectas como de captura, así como quimioluminiscencia y electroquimioluminiscencia.

Métodos

Se estudiaron 7 ensayos para rubéola y 6 para sarampión. Se emplearon 162 muestras (de 90 casos de rubéola y de 72 de sarampión) que se analizaron en todos los ensayos.

Resultados

Los rangos de sensibilidad, especificidad y concordancia para los ensayos de rubéola fueron 94,8-100%, 52,4-100% y 75,5-98,1%, respectivamente. Los rangos correspondientes para los ensayos de sarampión fueron 87-100%, 53,3-100% y 73-99,4%, respectivamente.

Conclusión

Los mejores ensayos fueron quimioluminiscencia (para IgM frente a rubéola y a sarampión) y electroquimioluminiscencia (para IgM frente a rubéola).

Palabras clave:
Virus rubéola
Virus sarampión
Inmunoensayo enzimático
Inmunoensayo quimioluminiscente
Inmunoensayo electroquimioluminiscente
Texto completo
Introduction

Rubella and measles infections are usually diagnosed in the laboratory by RT-PCR detection of the virus genomes during the first four days of the disease. However, samples taken at the appropriate time for making a direct diagnosis are not always available, so the detection of IgM-class antibodies is used as a fundamental tool for correct case characterization.1 Using a combination of the two approaches greatly improves the performance of measles2,3 and rubella2 diagnosis. In a situation of elimination, such as the current one, serological tests with good sensitivity and specificity characteristics are required. Siemens Healthcare Diagnostics (Marburg, Germany) (ELISA-Siemens) currently supplies the reference methods (ELISA assays, Enzygnost®) for determining IgM against rubella (RuV) and measles (MeV) viruses. The Enzygnost® kits have been used by a large number of network laboratories for many years.4,5 However, they will soon be withdrawn from the market, so the identification of alternative methods with acceptable performance characteristics is a challenge for clinical laboratories.

The aim of this study is to assess the operating characteristics of ELISA assays, based on indirect or μ chain capture methodologies, and chemiluminescence (CLIA) (capture), all for the purpose of determining the levels of IgM against RuV and MeV, and of electrochemiluminescence (ECLIA) (capture), for determining IgM versus RuV. Seven assays were compared for RuV IgM and six for MeV IgM.

Materials and methodsSerological methods

The following assays were studied (Table 1):

  • 1.

    Rubella assays

  • i.

    Enzygnost Rubella IgM, Siemens (Marburg, Germany) (RuV-ELISA-Siemens). The determinations were carried out in a Behring ELISA Processor III (BEPIII) (Siemens).

  • ii.

    Serion ELISA Classic Rubella IgM, Serion (Würzburg, Germany) (RuV-ELISA-Serion). The determinations were done in a BEPIII.

  • iii.

    Anti-Rubella IgM ELISA, EuroImmun (Lübeck, Germany) (RuV-ELISA-Euroimmun-1). The assay was done manually, and the plates were read in a BEPIII.

  • iv.

    Anti-Rubella IgM GP ELISA, EuroImmun (RuV-ELISA-Euroimmun-2). The tests were done manually, and the plates were read in a BEPIII.

  • v.

    Enzywell Rubella IgM, Diesse (Monteriggioni, Italy) (RuV-ELISA-Diesse). The tests were done manually, and the plates were read in a BEPIII.

  • vi.

    LIAISON® Rubella IgM, Diasorin (Saluggia, Italy) (RuV-CLIA). The determinations were done on a LIAISON® XL platform (Diasorin).

  • vii.

    ELECSYS® Rubella IgM, Roche Diagnostics (Penzberg, Germany) (RuV-ECLIA). The determinations were done in a Cobas e411 analyzer (Roche Diagnostics).

  • 2.

    Measles assays

  • i.

    Enzygnost Measles IgM, Siemens (MeV-ELISA-Siemens). The determinations were carried out in a BEPIII.

  • ii.

    Serion ELISA Classic Measles IgM, Serion (MeV-ELISA-Serion). The determinations were carried out in a BEPIII.

  • iii.

    Anti-Measles IgM ELISA, EuroImmun (MeV-ELISA-Euroimmun-1). The tests were done manually, and the plates were read in a BEPIII.

  • iv.

    Anti-Measles IgM NP ELISA, EuroImmun (MeV-ELISA-Euroimmun-2). Tests were done manually, and the plates were read in a BEPIII.

  • v.

    Enzywell Measles IgM, Diesse (MeV-ELISA-Diesse). Tests were done manually, and the plates were read in a BEPIII.

  • vi.

    LIAISON® Measles IgM (Diasorin) (MeV-CLIA). The determinations were done on a LIAISON® XL platform.

Table 1.

List of assays analyzed.

Assay  Manufacturer  Method  Platform/manually  Antigen  Format (no. samples)  Stability once opened  Samples tested (n
Rubella
Enzygnost Rubella IgMRuV-Siemens  Siemens  Indirect ELISA  BEPIII  Rubella-infected BHK21 cells  96  Expiry date  162 
ELISA Classic Rubella IgMRuV-Virion  Virion  Indirect ELISA  BEPIII  Rubella-infected cells (HPV-77)  96  Expiry date  162 
Anti-Rubella IgM ELISARuV-Euroimmun-1  Euroimmun  Indirect ELISA  Manually  Rubella-infected Vero cells (strain HPV-77)  96  Expiry date  162 
Anti-Rubella IgM GP ELISARuV-Euroimmun-2  Euroimmun 2  Indirect ELISA  Manually  Highly purified native GP (strain HPV-77, grown in Vero cells)  96  Expiry date  162 
Enzywell Rubella IgMRuV-Diesse  Diesse  Capture ELISA  Manually  Purified rubella virus  96  Expiry date  162 
LIAISON® Rubella IgMRuV-CLIA  Diasorin  Capture CLIA  LIAISON® XL  Rubella viral particle (strain HPV 77)  100  8 weeks  158 
Elecsys® Rubella IgMRuV-ECLIA  Roche  Capture ECLIA  Cobas e411  Rubella-like particles  100  14 days  160 
Measles
Enzygnost Measles IgMMeV-Siemens  Siemens  Indirect ELISA  BEPIII  Measles-infected Vero cells  96  Expiry date  162 
ELISA Classic Measles IgMMeV-Virion  Virion  Indirect ELISA  BEPIII  Measles-infected cells, strain Edmonston  96  Expiry date  162 
Anti-Measles IgM ELISAMeV-Euroimmun-1  Euroimmun  Indirect ELISA  Manually  Measles-infected cells, strain HNT-PI Vero cells  96  Expiry date  162 
Anti-Measles IgM NP ELISAMeV-Euroimmun-1  Euroimmun 2  Indirect ELISA  Manually  Recombinant NP expressed in eukaryotic cells  96  Expiry date  162 
Enzywell Measles IgMMeV-Diesse  Diesse  Capture ELISA  Manually  Purified measles virus