Development and evaluation of a capture ELISA for IgM antibody to the human cytomegalovirus major DNA binding protein

M. Grazia Revello, Elena Percivalle, Marco Zannino, Valdano Rossi, Giuseppe Gerna

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

A new capture ELISA (ELAb) for determination of the IgM antibody response to the human cytomegalovirus major DNA binding protein (p52) was developed by using a p52-specific monoclonal antibody. As a reference test, a capture ELISA using in parallel viral- and cell-control labeled antigens (ELA) was employed. General specificity, which was determined on 180 unselected IgM-negative sera from an adult population was 100%; stringent specificity, which was evaluated on 108 potentially interfering sera from patients with Epstein-Barr virus infectious mononucleosis, autoimmune diseases, rheumatoid factor or treated with radioimmunotherapy, was 96.3%; finally, clinical specificity, determined on 79 IgM-negative sera drawn prior to onset of primary HCMV infection, was 100%. Thus, the overall specificity was 98.9% ( 363 367) IgM negative tested sera). Sensitivity assayed on 277 IgM-positive sera was 100%. The study of the kinetics of the IgM antibody response in sequential blood samples from 9 immunocompetent and 9 heart transplanted patients showed that, while in the immunocompetent p52-specific IgM titer fell sharply 2-3 months after onset and was virtually undetectable 12 months after onset, in the immunocompromised the IgM response persisted for longer than a year. Recurrent HCMV infections were associated with a high titer IgM response in 6 (30%), and with a low IgM response in another 6 (30%) heart transplanted patients within a group of 20 patients sequentially examined. Finally, IgM antibodies were detected in all 4 infants with congenital infection and in 5 of 6 infants with neonatal infection. The results show that the HCMV p52-specific IgM antibody response parallels that obtained by ELA, thus representing a major component of it. ELAb is highly sensitive, specific and reproducible. It represents a major advance among capture ELISA techniques, allowing detection of IgM antibody reactive to a specific viral protein.

Original languageEnglish
Pages (from-to)315-329
Number of pages15
JournalJournal of Virological Methods
Volume35
Issue number3
DOIs
Publication statusPublished - 1991

Fingerprint

DNA-Binding Proteins
Cytomegalovirus
Immunoglobulin M
Enzyme-Linked Immunosorbent Assay
Antibodies
Antibody Formation
Serum
Infection
Radioimmunotherapy
Infectious Mononucleosis
Rheumatoid Factor
Viral Proteins
Human Herpesvirus 4
Autoimmune Diseases
Monoclonal Antibodies

Keywords

  • Capture ELISA
  • DNA binding protein
  • Human cytomegalovirus
  • IgM antibody
  • Monoclonal antibody

ASJC Scopus subject areas

  • Virology

Cite this

@article{3261451e6ac94095b701b3a2f6f24b4f,
title = "Development and evaluation of a capture ELISA for IgM antibody to the human cytomegalovirus major DNA binding protein",
abstract = "A new capture ELISA (ELAb) for determination of the IgM antibody response to the human cytomegalovirus major DNA binding protein (p52) was developed by using a p52-specific monoclonal antibody. As a reference test, a capture ELISA using in parallel viral- and cell-control labeled antigens (ELA) was employed. General specificity, which was determined on 180 unselected IgM-negative sera from an adult population was 100{\%}; stringent specificity, which was evaluated on 108 potentially interfering sera from patients with Epstein-Barr virus infectious mononucleosis, autoimmune diseases, rheumatoid factor or treated with radioimmunotherapy, was 96.3{\%}; finally, clinical specificity, determined on 79 IgM-negative sera drawn prior to onset of primary HCMV infection, was 100{\%}. Thus, the overall specificity was 98.9{\%} ( 363 367) IgM negative tested sera). Sensitivity assayed on 277 IgM-positive sera was 100{\%}. The study of the kinetics of the IgM antibody response in sequential blood samples from 9 immunocompetent and 9 heart transplanted patients showed that, while in the immunocompetent p52-specific IgM titer fell sharply 2-3 months after onset and was virtually undetectable 12 months after onset, in the immunocompromised the IgM response persisted for longer than a year. Recurrent HCMV infections were associated with a high titer IgM response in 6 (30{\%}), and with a low IgM response in another 6 (30{\%}) heart transplanted patients within a group of 20 patients sequentially examined. Finally, IgM antibodies were detected in all 4 infants with congenital infection and in 5 of 6 infants with neonatal infection. The results show that the HCMV p52-specific IgM antibody response parallels that obtained by ELA, thus representing a major component of it. ELAb is highly sensitive, specific and reproducible. It represents a major advance among capture ELISA techniques, allowing detection of IgM antibody reactive to a specific viral protein.",
keywords = "Capture ELISA, DNA binding protein, Human cytomegalovirus, IgM antibody, Monoclonal antibody",
author = "Revello, {M. Grazia} and Elena Percivalle and Marco Zannino and Valdano Rossi and Giuseppe Gerna",
year = "1991",
doi = "10.1016/0166-0934(91)90073-9",
language = "English",
volume = "35",
pages = "315--329",
journal = "Journal of Virological Methods",
issn = "0166-0934",
publisher = "Elsevier",
number = "3",

}

TY - JOUR

T1 - Development and evaluation of a capture ELISA for IgM antibody to the human cytomegalovirus major DNA binding protein

AU - Revello, M. Grazia

AU - Percivalle, Elena

AU - Zannino, Marco

AU - Rossi, Valdano

AU - Gerna, Giuseppe

PY - 1991

Y1 - 1991

N2 - A new capture ELISA (ELAb) for determination of the IgM antibody response to the human cytomegalovirus major DNA binding protein (p52) was developed by using a p52-specific monoclonal antibody. As a reference test, a capture ELISA using in parallel viral- and cell-control labeled antigens (ELA) was employed. General specificity, which was determined on 180 unselected IgM-negative sera from an adult population was 100%; stringent specificity, which was evaluated on 108 potentially interfering sera from patients with Epstein-Barr virus infectious mononucleosis, autoimmune diseases, rheumatoid factor or treated with radioimmunotherapy, was 96.3%; finally, clinical specificity, determined on 79 IgM-negative sera drawn prior to onset of primary HCMV infection, was 100%. Thus, the overall specificity was 98.9% ( 363 367) IgM negative tested sera). Sensitivity assayed on 277 IgM-positive sera was 100%. The study of the kinetics of the IgM antibody response in sequential blood samples from 9 immunocompetent and 9 heart transplanted patients showed that, while in the immunocompetent p52-specific IgM titer fell sharply 2-3 months after onset and was virtually undetectable 12 months after onset, in the immunocompromised the IgM response persisted for longer than a year. Recurrent HCMV infections were associated with a high titer IgM response in 6 (30%), and with a low IgM response in another 6 (30%) heart transplanted patients within a group of 20 patients sequentially examined. Finally, IgM antibodies were detected in all 4 infants with congenital infection and in 5 of 6 infants with neonatal infection. The results show that the HCMV p52-specific IgM antibody response parallels that obtained by ELA, thus representing a major component of it. ELAb is highly sensitive, specific and reproducible. It represents a major advance among capture ELISA techniques, allowing detection of IgM antibody reactive to a specific viral protein.

AB - A new capture ELISA (ELAb) for determination of the IgM antibody response to the human cytomegalovirus major DNA binding protein (p52) was developed by using a p52-specific monoclonal antibody. As a reference test, a capture ELISA using in parallel viral- and cell-control labeled antigens (ELA) was employed. General specificity, which was determined on 180 unselected IgM-negative sera from an adult population was 100%; stringent specificity, which was evaluated on 108 potentially interfering sera from patients with Epstein-Barr virus infectious mononucleosis, autoimmune diseases, rheumatoid factor or treated with radioimmunotherapy, was 96.3%; finally, clinical specificity, determined on 79 IgM-negative sera drawn prior to onset of primary HCMV infection, was 100%. Thus, the overall specificity was 98.9% ( 363 367) IgM negative tested sera). Sensitivity assayed on 277 IgM-positive sera was 100%. The study of the kinetics of the IgM antibody response in sequential blood samples from 9 immunocompetent and 9 heart transplanted patients showed that, while in the immunocompetent p52-specific IgM titer fell sharply 2-3 months after onset and was virtually undetectable 12 months after onset, in the immunocompromised the IgM response persisted for longer than a year. Recurrent HCMV infections were associated with a high titer IgM response in 6 (30%), and with a low IgM response in another 6 (30%) heart transplanted patients within a group of 20 patients sequentially examined. Finally, IgM antibodies were detected in all 4 infants with congenital infection and in 5 of 6 infants with neonatal infection. The results show that the HCMV p52-specific IgM antibody response parallels that obtained by ELA, thus representing a major component of it. ELAb is highly sensitive, specific and reproducible. It represents a major advance among capture ELISA techniques, allowing detection of IgM antibody reactive to a specific viral protein.

KW - Capture ELISA

KW - DNA binding protein

KW - Human cytomegalovirus

KW - IgM antibody

KW - Monoclonal antibody

UR - http://www.scopus.com/inward/record.url?scp=0025786521&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025786521&partnerID=8YFLogxK

U2 - 10.1016/0166-0934(91)90073-9

DO - 10.1016/0166-0934(91)90073-9

M3 - Article

C2 - 1667791

AN - SCOPUS:0025786521

VL - 35

SP - 315

EP - 329

JO - Journal of Virological Methods

JF - Journal of Virological Methods

SN - 0166-0934

IS - 3

ER -