scholarly journals Evaluation of Hemagglutinin Protein-Specific Immunoglobulin M for Diagnosis of Measles by an Enzyme-Linked Immunosorbent Assay Based on Recombinant Protein Produced in a High-Efficiency Mammalian Expression System

1998 ◽  
Vol 36 (12) ◽  
pp. 3509-3513 ◽  
Author(s):  
Fabienne B. Bouche ◽  
Nicolaas H. C. Brons ◽  
Sophie Houard ◽  
Francois Schneider ◽  
Claude P. Muller
Keyword(s):  
Recombinant Protein ◽  
Immunosorbent Assay ◽  
Measles Virus ◽  
High Efficiency ◽  
Expression System ◽  
Immunoglobulin M ◽  
Initial Time ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mammalian Expression ◽  
Specific Immunoglobulin

Recombinant hemagglutinin (H) of the measles virus (MV) expressed in a mammalian high-expression system based on the Semliki Forest virus replicon was used in an enzyme-linked immunosorbent assay (ELISA) for the detection of specific immunoglobulin M (IgM) and IgG in patients with acute-phase measles. One hundred twelve serum specimens from 70 patients with measles were analyzed. Case definition was based on a commercial IgM ELISA that utilizes MV-infected cells (MV-ELISA) (Enzygnost; Behring Diagnostics); the clinical criteria of the Centers for Disease Control and Prevention (Atlanta, Ga.); and/or the increase in hemagglutinin test titers, neutralization test titers, and levels of MV-specific IgG whenever paired sera were available. The initial time courses of the IgM signal after the onset of rash are similar in the H- and MV-ELISAs. On days 0 to 19, both ELISAs detected IgM in 67 of 68 (98.5%) sera. Average maximal levels of IgM seem to persist, however, about 10 days longer in the MV-ELISA (up to day 25) than in the H-ELISA (day 15). From days 20 to 29 and 30 to 59, the H-ELISA detected only 64.3 (9 of 14) and 19.2% (5 of 26), respectively, of sera that were IgM positive by MV-ELISA. At least up to day 30, the performance of the H-ELISA seemed to be similar to that reported for commercial ELISAs based on whole MV. Our results demonstrate that MV H-specific IgM can be used to diagnose most measles cases from a single serum specimen collected within 19 days after the onset of rash and that the recombinant protein used in this study is suitable for this purpose.

scholarly journals Immunosorbent Assay Based on Recombinant Hemagglutinin Protein Produced in a High-Efficiency Mammalian Expression System for Surveillance of Measles Immunity

1998 ◽  
Vol 36 (3) ◽  
pp. 721-726 ◽  
Author(s):  
Fabienne Bouche ◽  
Wim Ammerlaan ◽  
Francoise Berthet ◽  
Sophie Houard ◽  
Francois Schneider ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Mammalian Cells ◽  
Measles Virus ◽  
Expression System ◽  
False Negative ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Mammalian Expression ◽  
Recombinant Hemagglutinin ◽  
H Protein

Recombinant hemagglutinin (H) protein of the measles virus (MV) was produced in mammalian cells with a high-yield expression system based on the Semliki Forest virus replicon. Crude membrane preparations of H protein-transfected BHK-21 cells were used to coat microtiter plates to measure specific immunoglobulin G antibodies in 228 serologically defined serum samples mainly from measles late-convalescent adults. The titers by the enzyme-linked immunosorbent assay for the H protein (H-ELISA) closely correlated with neutralization test (NT) titers (R 2 = 0.66), hemagglutination inhibition test (HI) titers (R 2 = 0.64), with the titers from a certified commercial ELISA based on whole MV-infected cells (MV-ELISA;R 2 = 0.45). The correlations described above were better than those of the commercial MV-ELISA titers with the NT (R 2 = 0.52) or HI (R 2 = 0.48) titers. By using the 2nd International Standard for anti-measles serum, the detection level of the assay corresponds to 215 mIU/ml for undiluted serum, which corresponds to the estimated threshold for protective immunity. The specificity, accuracy, and positive predictive value were, in general, better for the H-ELISA than for a commercial MV-ELISA, independent of whether HI, NT, or HI and NT were used as “gold standards.” In contrast, the H-ELISA proved to be slightly less sensitive than the MV-ELISA (sensitivities, 98.6 versus 99.5%, respectively;P was not significant). The assays did not differ significantly in the number of serum samples with positive HI and NT results (n = 212) which measured false negative (H-ELISA, 2 of 212 [0.94%]; MV-ELISA, 1 of 212 [0.47%]), but the H-ELISA detected significantly more measles-susceptible individuals than the MV-ELISA (10 of 11 versus 3 of 11, respectively;P < 0.05) among the individuals whose sera had negative HI and NT results. Our data demonstrate that the H-protein preparation that we describe could be a cost-effective alternative to current whole-virus-based ELISAs for surveillance for immunity to measles and that such an assay could be more efficient in detecting susceptibility to measles. Furthermore, unlike whole MV-based antigens, H-protein would also be suitable for use in the development of a simple field test for the diagnosis of measles.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay for Immunoglobulin M Antibodies against Measles Virus

2003 ◽  
Vol 10 (3) ◽  
pp. 439-442 ◽  
Author(s):  
F. Roodbari ◽  
M. H. Roustai ◽  
A. Mostafaie ◽  
H. Soleimanjdahi ◽  
R. Sarrami Foroshani ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Neutralizing Antibodies ◽  
Measles Virus ◽  
Clinical Symptoms ◽  
Maculopapular Rash ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Vaccination Programs ◽  
Elisa System

ABSTRACT Measles is a highly contagious respiratory virus infection, with typical clinical symptoms including maculopapular rash, fever, cough, coryza, and conjunctivitis. Despite implementation of widespread vaccination programs throughout the world, the rates of global morbidity and mortality are still considerable. This study was performed to design a reliable indirect enzyme-linked immunosorbent assay (ELISA) to measure measles-specific immunoglobulin M (IgM). First, human IgM was purified, and then an anti-IgM antibody was produced in rabbits and purified in a multistep process. The rabbit IgG against human IgM was conjugated with peroxidase. Measles virus-infected Vero cells produced viral antigen. One hundred serum samples from infants of 9 to 18 months of age, mostly vaccinated, were evaluated for determining the presence of specific IgM antibodies against measles virus. The samples were also evaluated for neutralizing antibodies against measles virus by a microneutralization test (MNT). By comparing the results of the ELISA with those of MNT, it was demonstrated that ELISA had a sensitivity and specificity of 100 and 92%, respectively. On the other hand, when the results obtained by our ELISA system were compared with those of an imported measles virus IgM ELISA kit (EIAgen; Adaltis Italia SPa, Bologna, Italy), a high level of agreement was shown (k = 0.926).

scholarly journals Lateral-Flow Assay for Rapid Serodiagnosis of Human Leptospirosis

2001 ◽  
Vol 8 (1) ◽  
pp. 166-169 ◽  
Author(s):  
Henk L. Smits ◽  
C. K. Eapen ◽  
Sheela Sugathan ◽  
Mariamma Kuriakose ◽  
M. Hussein Gasem ◽  
...  
Keyword(s):  
Positive Result ◽  
Immunosorbent Assay ◽  
Rapid Detection ◽  
Test Line ◽  
Lateral Flow ◽  
Immunoglobulin M ◽  
Lateral Flow Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Immunoglobulin ◽  
Human Sera

ABSTRACT An assay device for the rapid detection ofLeptospira-specific immunoglobulin M (IgM) antibodies in human sera is presented. The sensitivity (85.8%) and specificity (93.6%) of the assay compared well (91.9% agreement) with those of an IgM enzyme-linked immunosorbent assay routinely used in the serodiagnosis of leptospirosis. The sensitivity of the assay varied with the stage of the disease. The assay uses stabilized components and is simply performed by the addition of serum and sample fluid to the sample well of the assay device. The assay is read after 10 min, and a positive result is obtained when staining of the test line is observed.

scholarly journals Altered Enzyme-Linked Immunosorbent Assay Immunoglobulin M (IgM)/IgG Optical Density Ratios Can Correctly Classify All Primary or Secondary Dengue Virus Infections 1 Day after the Onset of Symptoms, when All of the Viruses Can Be Isolated

2006 ◽  
Vol 13 (9) ◽  
pp. 1044-1051 ◽  
Author(s):  
Andrew K. I. Falconar ◽  
Elsa de Plata ◽  
Claudia M. E. Romero-Vivas
Keyword(s):  
Dengue Virus ◽  
Optical Density ◽  
Immunosorbent Assay ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Virus Infections ◽  
Igm Elisa ◽  
Elisa Titer ◽  
Specific Immunoglobulin ◽  
Density Ratios

ABSTRACT We compared dengue virus (DV) isolation rates and tested whether acute primary (P) and acute/probable acute secondary (S/PS) DV infections could be correctly classified serologically when the patients' first serum (S1) samples were obtained 1 to 3 days after the onset of symptoms (AOS). DV envelope/membrane protein-specific immunoglobulin M (IgM) capture and IgG capture enzyme-linked immunosorbent assay (ELISA) titrations (1/log10 1.7 to 1 log10 6.6 dilutions) were performed on 100 paired S1 and S2 samples from suspected DV infections. The serologically confirmed S/PS infections were divided into six subgroups based on their different IgM and IgG responses. Because of their much greater dynamic ranges, IgG/IgM ELISA titer ratios were more accurate and reliable than IgM/IgG optical density (OD) ratios recorded at a single cutoff dilution for discriminating between P and S/PS infections. However, 62% of these patients' S1 samples were DV IgM and IgG titer negative (<ODmax/2 titer threshold), and in 35% of the S/PS infections, the patients' S1 and S2 samples were IgM titer negative. The IgM OD values were, however, much higher than those of IgG in the S1 samples of many of these, and the other, S/PS infections. This necessitated using higher (≥2.60 and <2.60) discriminatory IgM/IgG OD (DOD) ratios on these S1 samples than those published previously to correctly classify the highest percentage of these P and S/PS infections. The DV isolation rate was highest (12/12; 100%) using IgG and IgM titer-negative S1 samples collected 1 day AOS, when 100% of them were correctly classified as P or S/PS infections using these higher DOD ratios.

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