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Journal of Clinical Microbiology, December 1998, p. 3509-3513, Vol. 36, No. 12
Laboratoire National de Santé, L-1011
Luxembourg, Luxembourg1;
Service de
Génétique Appliquée, Université Libre de
Bruxelles, B-1400 Nivelles, Belgium2; and
Medizinische Fakultät, Universität
Tübingen, D-72076 Tübingen, Germany3
Received 18 May 1998/Returned for modification 29 June
1998/Accepted 18 September 1998
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.
Vaccination has reduced the
worldwide morbidity and mortality of measles, and eradication of the
disease within the next few decades has become a realistic goal.
Despite high vaccination coverage, outbreaks continue to occur, and it
is likely that a rapid intervention strategy after detection of measles
cases will be required. However, rare and isolated cases tend to become
more difficult to diagnose clinically. Incomplete protection after vaccination (vaccine-modified infections) can result in clinical measles with uncharacteristic symptoms (9, 22). Numerous diseases with similar skin involvement, such as allergies, add to the
difficulties of measles diagnosis. These difficulties are compounded in
patients with dark skin. Therefore, measles diagnosis relies
increasingly on serological tests.
Diagnosis of measles may be confirmed by virus isolation, by the
demonstration of a significant increase in specific immunoglobulin G
(IgG) titers, or by the detection of anti-measles virus (MV) IgM
antibodies by using radioimmunoassays (2, 18, 37), enzyme-linked immunosorbent assays (ELISAs) (27, 34), and direct or indirect fluorescence-antibody techniques (20,
24). MV IgM appears at the same time as rash (12, 21,
34) and can be detected 3 days after the onset of rash in most
individuals (29, 32). IgM peaks on days 7 to 10 and wanes
within weeks (28). Since IgM is transient, the demonstration
of specific IgM corresponds to a recent primary measles infection
(14, 21). A single serum specimen collected at the
appropriate time (14) is now generally accepted to be
sufficient to diagnose measles (10, 12, 14, 17, 26, 32, 34,
37).
Current IgG and IgM assays are based on whole MV or virus-infected
cells as antigens (8, 10, 11, 33, 35, 36, 40). These
antigens are possibly more costly and difficult to produce and preserve
under standardized conditions than are recombinant proteins.
Rapid diagnosis of measles is essential for the timely implementation
of control measures to prevent the spread of infection. Inexpensive,
simple, and rapid tests which could be used under field conditions, as
an alternative to whole-virus-based ELISA, would represent an important
step towards measles control. Assays based on recombinant proteins
would potentially benefit from the higher stability of their antigen.
The nucleoprotein has been described as a suitable antigen to detect
specific IgG and IgM (16, 38, 39). More recently we have
shown that recombinant hemagglutinin (H) protein produced in a
high-yield mammalian expression system can be used for the surveillance
of measles immunity in late convalescents (3) and in
vaccinees (4). The high sensitivity and specificity of this
assay was due to high levels of detectable H-specific IgG antibodies in
both of these cohorts.
In the present study, we have investigated whether this recombinant
assay could also be used for detection of H-specific IgM antibodies and
for the diagnosis of measles.
(This work was done by Fabienne B. Bouche in partial fulfillment of the
requirements for a Ph.D. from Université Libre de Bruxelles,
Brussels, Belgium.)
Serum panel.
From 1996 to 1997, 112 serum samples were
collected from 70 patients (age range, 1.1 to 35.4 years; median, 8.2 years) during a major outbreak and from several isolated cases of
measles in Luxembourg. Paired sera were obtained from 31 patients; 4 patients were bled three times, and 1 patient was bled four times
within 59 days after the onset of rash. A total of 70 first samples (50 on or before day 19), 36 second samples (17 on or before day 19), 5 third samples (4 on or before day 19), and 1 fourth sample (before day
19) were collected. All patients had confirmed cases of measles, based
on the detection of specific IgM antibodies in a certified commercial
ELISA (MV-ELISA) (Enzygnost; Behring Diagnostics, Marburg, Germany). In
some patients measles was also confirmed by an increase in specific
antibodies in paired sera (n = 36) and/or the clinical case definition of the Centers for Disease Control and Prevention (CDC)
(Atlanta, Ga.) (n = 24) (Fig.
1). The CDC criteria include (i)
generalized maculopapular rash for 3 days or more; (ii) fever of
38.3°C, if measured; (iii) and at least one of the following symptoms; cough, coryza, or conjunctivitis (7). All patients presented with a typical rash. Four of the 112 samples were obtained between 2 and 14 days before the onset of rash; these cases were also
confirmed by increased hemagglutination inhibition (HI) and neutralization (NT) test titers and by MV-specific IgG and IgM antibodies in paired samples drawn after the onset of rash.
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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
ABSTRACT
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
INTRODUCTION
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
MATERIALS AND METHODS
Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
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FIG. 1.
Case definition by IgM in MV-ELISA, by CDC criteria, or
by increase in HI, NT, and specific IgG levels (found in all paired
sera) of 70 patients with measles from whom single or paired (or
multiple) sera were available. For most patients, CDC criteria were not
evaluated. (A) and (B) correspond to patients A and B in Fig. 2.
The numbers represent the numbers of serum samples.
Serological assays.
Anti-MV IgG and IgM were measured by
using a certified commercial MV-ELISA (Enzygnost for IgG and IgM;
Behring Diagnostics) based on whole-MV-infected simian cells, following
the supplier's instructions. HI and NT titers were determined as
described before (15). The titers are expressed as
log2 dilutions, with values of 
1:24
considered negative.
H-ELISA. (i) Antigen. MV H protein was obtained from a crude membrane preparation of BHK-21 cells transiently expressing the protein, as previously described (3). The negative control antigen was extracted from untransfected or mock-transfected BHK-21 cells. These preparations were used as antigens to determine IgG and IgM values in the H-ELISA.
(ii) IgM detection. Microtiter plates (Maxisorp; Nunc, Roskilde, Denmark) were coated with 50 µl of a mixture of three conformation-dependent H-specific monoclonal antibodies (5 µg/ml) in 0.1 M sodium bicarbonate buffer (pH 9.6). The monoclonal antibodies were derived from mice immunized with Edmonston strain MV. The plates were washed three times with 1% Tween 20 in Tris-buffered saline (15 mM; pH 8.0) and incubated for 75 min at room temperature with 50 µl of the above H antigen/well (10 µg of protein/ml) or negative control antigen. The plates were blocked with 1% bovine serum albumin in Tris-buffered saline (15 mM; pH 7.4). Test sera were diluted 1:10 in GULLSORB (Gull Laboratories, Louvain-La Neuve, Belgium) to eliminate interference by IgG. The sera were further diluted to a final concentration of 1:25 in a modified commercial dilution buffer (Enzymum-test; Boehringer, Mannheim, Germany) and added for 75 min at room temperature to the antigen-coated microtiter plates. The plates were washed three times with the above-mentioned wash buffer. Alkaline phosphatase-conjugated goat anti-human IgM (1:1,000; Southern Biotechnology Associates, Birmingham, Ala.) and p-nitrophenylphosphate (0.5 mg/ml; 100 µl/well) (Sigma, St. Louis, Mo.) were used to develop the assay. Optical density (OD) was measured at 405 nm following a 2-h incubation at 37°C. Data are expressed as milli-OD (mOD). The threshold for positivity was defined as the mean mOD (+ 2 standard deviations [SD]) of the IgM-negative sera, measured after 2 h.
(iii) IgG detection. Specific IgG was measured by H-ELISA following the procedure described previously (3). In this assay, the H antigen and the negative control antigen were directly coated onto microtiter plates. The threshold for positivity (210 mOD) was defined with sera that were negative for HI and NT, as described previously (3).
For IgM and IgG detection, OD was measured after 2 h at 405 nm, and data are expressed as net mOD by subtracting for each serum sample the background obtained with the control antigen from the H antigen.Statistics. The data were analyzed with Sigmastat software (Jandel Scientific, Erkrath, Germany). The z test was applied to compare the proportions of two groups of values within a cohort, and the t test was used to evaluate the significance of the difference between the mean values of two groups.
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RESULTS |
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Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
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Definition of threshold for positivity.
Specific IgM was
measured by H-ELISA with sera obtained from late-convalescent adults
with measles and from vaccinees; all of these individuals were negative
for IgM by MV-ELISA. The IgG levels of these donors were between 270 and 2,750 mOD by MV-ELISA. By H-ELISA, the means (± SD) of IgM values
for the convalescents and the vaccinees were 39 ± 61 (range, 
77
to 131) and 23 ± 49 (range, 45 to 105), respectively. The
t test detected no significant difference between the means
of vaccinees and convalescents. When the IgM values of sera with IgG
levels of <1,000 mOD and of those with >1,000 mOD were compared,
no difference was detectable (26 ± 58.2 versus 35 ± 57.2; P value, not significant).
Detection of MV-specific IgM in patients with measles.
In 108 serum samples (from 70 different patients) drawn at different time
intervals after the onset of rash, levels of specific IgM were analyzed
both by H-ELISA and by MV-ELISA. Figure 2
shows that the MV-ELISA detected 107 IgM-positive sera (99.1%) whereas the H-ELISA detected IgM in only 81 sera (75%) on the basis of the
above threshold. All (additional) sera collected before the onset of
rash (on days 14, 9, 8, and 2; n = 4) were IgM
negative in both ELISAs (Fig. 3).
|
) and on days 0 to 19 (
) and days 20 to 59 (
) after the onset of rash (n = 108).
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Detection of IgG by H-ELISA in patients with measles. With the MV-ELISA, IgG could be detected in all samples after day 3 (i.e., no false-negative sample). When the first serum sample (n = 16) was drawn between days 0 and 15, the second serum sample (drawn 7 to 54 days later) exhibited a mean increase of 1,013 mOD (range, 146 to 2,076 mOD) in the H-ELISA and 959 mOD (range, 215 to 1,963 mOD) in the MV-ELISA. Thus, no significant difference between the two assays became apparent (Fig. 4).
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DISCUSSION |
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Top
Abstract
Introduction
Materials & Methods
Results
Discussion
References
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Currently the most reliable diagnostic assays for measles are based on the detection of IgM by ELISA with whole MV or MV-infected cells as antigens (5, 10, 40). ELISAs based on recombinant proteins detect only a fraction of the total MV antibodies. The time of appearance of protein-specific antibodies is critical for the sensitivity of the assay early after the onset of rash. Nucleoprotein-specific antibodies may appear somewhat earlier during measles infection and are thought to be the most abundant antibodies early after the onset of rash (13, 25, 27), but this protein shows considerable sequence variability (31). In contrast, lower levels of H-specific antibodies were detected by immunoprecipitation (13), by competition ELISA with H-specific monoclonal antibodies (27), and by comparing complement fixation with immunodiffusion tests (25). In addition, these antibodies seemed to develop only later after the onset of rash (13). These observations suggested that H-specific antibodies may not be sufficient for diagnosis, but the observations are in conflict with the rapid increase of HI and NT antibodies, both of which are predominantly directed against H protein after the onset of rash (reference 30 and unpublished results).
Our study demonstrates that there are considerable amounts of H-specific antibodies within the early days after the onset of rash which are readily detectable with an ELISA based on a mammalian expressed recombinant protein. The initial time courses of IgM in the H-ELISA and the commercial MV-ELISA seemed to be similar. Maximal average levels of specific IgM were reached between days 5 and 10 after the onset of rash. This is in agreement with earlier studies with total IgM isolated by sucrose gradient and tested by HI (day 10) (6) or by IgM Enzygnost (days 5 to 10) (28, 32).
The early appearance of H-specific IgM translates into a high sensitivity (98.5%) of the H-ELISA within the first 19 days, which matches that of the Enzygnost MV-ELISA (98.5%) for the same sera. When an interval from 0 to 30 days is considered, the sensitivity of the H-ELISA was 92.7% and that of the MV-ELISA was 98.8%. In another cohort the sensitivity between days 0 and 30 was reported to be 91.8% for the Behring test (specificity, 98.2%), 93.3% for the Gull test (specificity, 90.5%), and 85.5% for the Incstar test (specificity, 95.2%) (1). When an even longer interval is considered, the sensitivity of the H-ELISA rapidly deteriorated (days 0 to 59, 75%) while the sensitivity of the MV-ELISA did not (98.1%) because it served as a "gold standard."
The high sensitivity seems to agree with the results of Ozanne and d'Halewyn (26), who reported a sensitivity of 91.9% for Enzygnost between days 1 and 56. However, the overall sensitivity strongly depends on the number of sera collected during the later time points. In some studies only 0, 8, 5, or 4.5% of the samples were collected after day 15 (32), day 19 (23), day 25 (26), or day 30 (1), respectively. In contrast, 37% of the samples in our study were drawn after day 20 and 25% of the samples were collected after day 30. It is therefore important to compare sensitivities later after the onset of rash. In the H-ELISA, the sensitivity was 64.3 (n = 14) and 19.2% (n = 26) between days 20 and 29 and days 30 and 59, respectively. In the study of Arista et al. (1), the sensitivity of Enzygnost was 40% between days 31 and 35.
After day 19, less than half of the sera (16 of 40) were solely confirmed by MV-ELISA. Therefore, the assessment of the sensitivity of the H-ELISA after day 19 may depend on the performance of IgM Enzygnost. However, essentially no difference in the sensitivity of the H-ELISA was found whether the sera were confirmed by the commercial ELISA only (29.4%) or by at least one additional parameter (37.5%). Thus, in our cohort, the difference in sensitivity between the H-ELISA and the MV-ELISA cannot be explained by excessive false-positive results (low specificity) of the MV-ELISA but rather by the accelerated waning of H-specific IgM in comparison to that of MV IgM.
The threshold that gave a sensitivity of 98.8% in the H-ELISA (days 0 to 19) was associated with a specificity of 100% in the IgM-negative panel. However, this needs to be confirmed in a panel of IgM-negative sera which is independent of the definition of the threshold. Lowering the threshold of the H-ELISA would decrease the specificity but increase the sensitivity above 95% within the first 30 days. A high specificity, i.e., a low percentage of false-positive results, was also reported by several authors for the Behring test (1, 26, 32).
Measles is most contagious within 1 week before and 1 week after the onset of rash (19). Serological assays before the onset of rash are not available, but during the contagious period after the onset of rash the H-ELISA and the MV-ELISA perform equally well. Most studies which rely on IgM for measles diagnosis recommend that serum samples be drawn within about 3 weeks after the onset of rash (1, 10, 14, 21, 26, 32). Thus, an optimal performance within 19 days seems to be sufficient for measles diagnosis (14, 26). Nonetheless, we are currently investigating whether further optimizing the assay could increase the sensitivity beyond day 19.
We have also studied the development of H-specific IgG in patients with measles. No false-negative serum was detected, and increases between paired sera were as significant as they were with MV-ELISA. In light of these results and the overall high sensitivity and specificity of H-ELISA for IgG (3, 4), we conclude that this assay is as reliable for the diagnosis of measles in paired sera as an ELISA based on whole MV. Despite the longer persistence of maximal levels of IgM in MV-ELISA and the early waning of H-specific IgM, the initial time courses of H- and MV-specific IgGs are similar.
Since the H protein can be efficiently produced in a mammalian system (3, 4), and since recombinant proteins may benefit from enhanced stability, an ELISA based on this antigen seems like an interesting alternative in the search for a low-cost, rapid diagnostic system for measles.
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ACKNOWLEDGMENTS |
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We thank the children with measles and their parents, their teachers, and their doctors, and in particular E. Mertens (Clervaux) for excellent collaboration and W. Ammerlaan for excellent technical help during the production and characterization of the recombinant measles H protein.
F.B.B. was supported by a fellowship from the Ministère de l'Education Nationale. This study was also supported by a grant from the Centre de Recherche Public-Santé, Luxembourg (CRP93/08).
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FOOTNOTES |
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* Corresponding author. Mailing address: Department of Immunology, Laboratoire National de Santé, 20A, rue Auguste Lumière, L-1011 Luxembourg, Luxembourg. Phone: 00352-490604. Fax: 00352-490686. E-mail: claude.muller{at}santel.lu.
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REFERENCES |
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Abstract
Introduction
Materials & Methods
Results
Discussion
References
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| 1. | Arista, S., D. Ferraro, A. Cascio, E. Vizzi, and R. di Stefano. 1995. Detection of IgM antibodies specific for measles virus by capture and indirect enzyme immunoassays. Res. Virol. 146:225-232[Medline]. |
| 2. |
Arstila, P.,
T. Vuorimaa,
K. Kalimo,
P. Halonen,
M. Viljanen,
K. Granfors, and P. Toivanen.
1977.
A solid-phase radioimmunoassay for IgG and IgM antibodies against measles virus.
J. Gen. Virol.
34:167-176 |
| 3. |
Bouche, F.,
W. Ammerlaan,
F. Berthet,
S. Houard,
F. Schneider, and C. P. Muller.
1998.
Immunosorbent assay based on recombinant hemagglutinin protein produced in a high-efficiency mammalian expression system for surveillance of measles immunity.
J. Clin. Microbiol.
36:721-726 |
| 4. | Bouche, F., W. Ammerlaan, F. Schneider, and C. P. Muller. 1998. A simplified immunoassay based on measles virus recombinant hemagglutinin for testing the immune status of vaccinees. J. Virol. Methods 74:77-87[Medline]. |
| 5. |
Boteler, W. L.,
P. M. Luipersbeck,
D. A. Fuccillo, and A. J. O'Beirne.
1983.
Enzyme-linked immunosorbent assay for detection of measles antibody.
J. Clin. Microbiol.
17:814-818 |
| 6. | Centers for Disease Control. 1982. Serologic diagnosis of measles. Morbid. Mortal. Weekly Rep. 31:396-402[Medline]. |
| 7. | Centers for Disease Control. 1983. Classification of measles cases and categorization of measles elimination programs. Morbid. Mortal. Weekly Rep. 31:707-711[Medline]. |
| 8. |
Cremer, N. E.,
C. K. Cossen,
G. Shell,
J. Diggs,
D. Gallo, and N. J. Schmidt.
1985.
Enzyme immunoassay versus plaque neutralization and other methods for determination of immune status to measles and varicella-zoster viruses and versus complement fixation for serodiagnosis of infections with those viruses.
J. Clin. Microbiol.
21:869-874 |
| 9. |
Edmonson, M. B.,
D. G. Addiss,
J. T. McPherson,
J. L. Berg,
S. R. Circo, and J. P. Davis.
1990.
Mild measles and secondary vaccine failure during sustained outbreak in a highly vaccinated population.
JAMA
263:2467-2471 |
| 10. |
Erdman, D. D.,
L. J. Anderson,
D. D. Adams,
J. A. Stewart,
L. E. Markowitz, and W. J. Bellini.
1991.
Evaluation of monoclonal antibody-based capture enzyme immunoassays for detection of specific antibodies to measles virus.
J. Clin. Microbiol.
29:1466-1471 |
| 11. | Erdman, D. D., J. L. Heath, J. C. Watson, L. E. Markowitz, and W. J. Bellini. 1993. Immunoglobulin M antibody response to measles virus following primary and secondary vaccination and natural virus infection. J. Med. Virol. 41:44-48[Medline]. |
| 12. |
Forghani, B.,
C. K. Myoraku, and N. J. Schmidt.
1983.
Use of monoclonal antibodies to human immunoglobulin M in "capture" assays for measles and rubella immunoglobulin M.
J. Clin. Microbiol.
18:652-657 |
| 13. |
Graves, M.,
D. E. Griffin,
R. T. Johnson,
R. L. Hirsch,
I. L. de Soriano,
S. Roedenbeck, and A. Vaisberg.
1984.
Development of antibody to measles virus polypeptides during complicated and uncomplicated measles virus infection.
J. Virol.
49:409-412 |
| 14. | Helfand, R. F., J. L. Heath, L. J. Anderson, E. F. Maes, D. Guris, and W. J. Bellini. 1997. Diagnosis of measles with an IgM capture EIA: the optimal timing of specimen collection after rash onset. J. Infect. Dis. 175:195-199[Medline]. |
| 15. | Huiss, S., B. Damien, F. Schneider, and C. P. Muller. 1997. Characteristics of asymptomatic secondary immune responses to measles virus in late convalescent donors. Clin. Exp. Immunol. 109:416-420[Medline]. |
| 16. |
Hummel, K. B.,
D. D. Erdman,
J. Heath, and W. J. Bellini.
1992.
Baculovirus expression of the nucleoprotein gene of measles virus and utility of the recombinant protein in diagnostic enzyme immunoassays.
J. Clin. Microbiol.
30:2874-2880 |
| 17. |
James, K.
1990.
Immunoserology of infectious diseases.
Clin. Microbiol. Rev.
3:132-152 |
| 18. | Jankowski, M. A., W. Gut, and M. Kantoch. 1982. Radioimmunoassay of measles virus antibodies in SSPE. Acta Virol. 26:481-487[Medline]. |
| 19. | Katz, S. L. 1985. Measles and subacute sclerosing panencephalitis, 18th ed., p. 1062. Appleton-Century-Crofts, New York, N.Y. |
| 20. | Kleiman, M. B., C. A. Blockhold, and N. J. Schmidt. 1986. Use of monoclonal antibodies to human immunoglobulin M in "capture" assays for measles and rubella immunoglobulin M. J. Clin. Microbiol. 18:652-657. |
| 21. |
Lievens, A. W., and P. A. Brunell.
1986.
Specific immunoglobulin M enzyme-linked immunosorbent assay for confirming the diagnosis of measles.
J. Clin. Microbiol.
24:391-394 |
| 22. | Makkhene, M. K., and P. S. Diaz. 1993. Clinical presentations and complications of suspected measles in hospitalized children. Pediatr. Infect. Dis. 12:836-840. |
| 23. |
Mayo, D. R.,
T. Brennan,
D. P. Cormier,
J. Hadler, and P. Lamb.
1991.
Evaluation of a commercial measles virus immunoglobulin M enzyme immunoassay.
J. Clin. Microbiol.
29:2865-2867 |
| 24. | Minnich, L., and C. G. Ray. 1980. Comparison of direct immunofluorescent staining of clinical specimens for respiratory virus antigens with conventional isolation techniques. J. Clin. Microbiol. 12:2865-2867. |
| 25. |
Norrby, E., and Y. Gollmar.
1972.
Appearance and persistence of antibodies against different virus components after regular measles infections.
Infect. Immun.
6:240-247 |
| 26. |
Ozanne, G., and M.-A. d'Halewyn.
1992.
Performance and reliability of the Enzygnost measles enzyme-linked immuno-sorbent assay for detection of measles virus-specific immunoglobulin M antibody during a large measles epidemic.
J. Clin. Microbiol.
30:564-569 |
| 27. | Pedersen, I. R., A. Antonsdottir, T. Evald, and C. H. Mordhorst. 1982. Detection of measles IgM antibodies by enzyme-linked immunosorbent assay (ELISA). Acta Pathol. Microbiol. Immunol. Scand. Sect. B 90:153-160[Medline]. |
| 28. | Pederson, I. R., C. H. Mordhorst, T. Ewald, and H. von Magnus. 1986. Long-term antibody response after measles vaccination in an isolated arctic society in Greenland. Vaccine 4:173-178[Medline]. |
| 29. | Perry, K. R., D. W. G. Brown, J. V. Parry, S. Panday, C. Pipkin, and A. Richards. 1993. Detection of measles, mumps, and rubella antibodies in saliva using antibody capture radioimmunoassay. J. Med. Virol. 40:235-240[Medline]. |
| 30. | Preblud, S. R., and S. L. Katz. 1988. Measles vaccine, p. 182-222. In S. A. Plotkin, and E. A. Mortimer, Jr. (ed.), Vaccines. W. B. Saunders, Philadelphia, Pa. |
| 31. | Rima, B. K., R. G. Wishaupt, M. J. Welsh, and J. A. Earle. 1995. The evolution of morbilliviruses: a comparison of nucleocapsid gene sequences including a porpoise morbillivirus. Vet. Microbiol. 44:127-134[Medline]. |
| 32. |
Rossier, E.,
H. Miller,
B. McCulloch,
L. Sullivan, and K. Ward.
1991.
Comparison of immunofluorescence and enzyme immunoassay for detection of measles-specific immunoglobulin M antibodies.
J. Clin. Microbiol.
29:1069-1071 |
| 33. | Salonen, J., R. Vainionpaa, and P. Halonen. 1986. Assay of measles virus IgM and IgG class antibodies by use of peroxidase-labelled viral antigens. Arch. Virol. 29:93-106. |
| 34. | Tuokko, H., and A. Salmi. 1983. Detection of IgM antibodies to measles virus by enzyme-immunoassay. Med. Microbiol. Immunol. 171:187-198[Medline]. |
| 35. | Tuokko, H. 1988. The detection of measles specific immunoglobulin M antibodies using biotinylated antigens. APMIS 96:491-496[Medline]. |
| 36. | Vaz De Lima, L. R., S. Hoshino-Shimizu, V. A. de Souza, C. S. Pannuti, H. F. Andrade, Jr., L. M. Sumita, and A. W. Ferreira. 1994. Measles serodiagnosis: standardization and evaluation of a Dot-ELISA. Rev. Inst. Med. Trop. Sao Paulo 36:139-147[Medline]. |
| 37. | Vuorimaa, T. O., P. P. Aristila, B. R. Ziola, A. A. Salmi, P. T. Hanninen, and P. E. Halonen. 1978. Solid phase radioimmunoassay determination of virus specific IgM antibody levels in a follow-up of patients with naturally acquired measles infections. J. Med. Virol. 2:271-278[Medline]. |
| 38. | Warnes, H., A. R. Fooks, and J. R. Stephenson. 1994. Production of measles nucleoprotein in different expression systems and its use as a diagnostic reagent. J. Virol. Methods 49:257-268[Medline]. |
| 39. | Warnes, A., A. R. Fooks, A. B. Dowsett, G. W. G. Wilkinson, and J. R. Stephenson. 1995. Expression of the measles virus nucleoprotein gene in Escherichia coli and assembly of nucleocapsid-like structures. Gene 160:173-178[Medline]. |
| 40. |
Weigle, K. A.,
M. D. Murphy, and P. A. Brunell.
1984.
Enzyme-linked immunosorbent assay for evaluation of immunity to measles virus.
J. Clin. Microbiol.
19:376-379 |
Journal of Clinical Microbiology, December 1998, p. 3509-3513, Vol. 36, No. 12
0095-1137/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
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