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Article ; Online: Measles IgG Avidity Assay.

Mercader, Sara / Crooke, Stephen

Methods in molecular biology (Clifton, N.J.)

2024 Volume 2808, Page(s) 247–264

Abstract: Measles IgG avidity assays determine the overall strength of molecular binding between measles-specific IgG antibodies and measles virus antigens. Avidity results can distinguish recent from distant measles virus infections. Individuals who are ... ...

Abstract	Measles IgG avidity assays determine the overall strength of molecular binding between measles-specific IgG antibodies and measles virus antigens. Avidity results can distinguish recent from distant measles virus infections. Individuals who are immunologically naïve to measles virus develop low-avidity antibodies upon measles virus infection or first-time vaccination. Within 4-6 months, antibodies mature to high avidity. Measles avidity assays are most useful in the context of measles elimination. In such settings, avidity and epidemiological and clinical information are used to classify measles breakthrough infections for control and surveillance purposes and to assist in case confirmation when other laboratory results are inconclusive or nonexistent. We present a highly accurate end-titer measles avidity assay that delivers results based on IgG quality (avidity) that are independent of IgG concentration.
MeSH term(s)	Antibody Affinity/immunology ; Immunoglobulin G/immunology ; Humans ; Antibodies, Viral/immunology ; Measles virus/immunology ; Measles/immunology ; Measles/virology ; Antigens, Viral/immunology ; Enzyme-Linked Immunosorbent Assay/methods
Language	English
Publishing date	2024-05-14
Publishing country	United States
Document type	Journal Article
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-0716-3870-5_18
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Article ; Online: Performance Characteristics of Six Immunoglobulin M Enzyme-Linked Immunosorbent Assays Used for Laboratory Confirmation of Measles.

Sowers, Sun B / Anthony, Kiana / Mercader, Sara / Colley, Heather / Crooke, Stephen N / Rota, Paul A / Latner, Donald R / Hickman, Carole J

Journal of clinical microbiology

2022 Volume 60, Issue 12, Page(s) e0122722

Abstract: Laboratory confirmation of infection is an essential component of measles surveillance. Detection of measles-specific IgM in serum by enzyme-linked immunosorbent assay (ELISA) is the most common method used to confirm measles infection. ELISA formats ... ...

Abstract	Laboratory confirmation of infection is an essential component of measles surveillance. Detection of measles-specific IgM in serum by enzyme-linked immunosorbent assay (ELISA) is the most common method used to confirm measles infection. ELISA formats vary, as does the sensitivity and specificity of each assay. Specimens collected within 3 days of rash onset can yield a false-negative result, which can delay confirmation of measles cases. Interfering substances can yield a false-positive result, leading to unnecessary public health interventions. The IgM capture assay developed at the Centers for Disease Control (CDC) was compared against five commercially available ELISA kits for the ability to detect measles virus-specific IgM in a panel of 90 well-characterized specimens. Serum samples were tested in triplicate using each commercial kit as recommended by the manufacturer. Using the CDC measles IgM capture assay as the reference test; the sensitivity and specificity for each commercial kit ranged from 50 to 83% and 86.9 to 98%, respectively. Discrepant results were observed for samples tested with all five commercial kits and ranged from 13.8 to 28.8% of the specimens tested. False-positive results occurred in 2.0 to 13.1% of sera, while negative results were observed in 16.7 to 50% of sera that were positive by the CDC measles IgM capture assay. Evaluation and interpretation of measles IgM serologic results can be complex, particularly in measles elimination settings. The performance characteristics of a measles IgM assay should be carefully considered when selecting an assay to achieve high-quality measles surveillance.
MeSH term(s)	Humans ; Immunoglobulin M ; Enzyme-Linked Immunosorbent Assay/methods ; Measles/diagnosis ; Measles/epidemiology ; Measles virus ; Sensitivity and Specificity ; Antibodies, Viral
Chemical Substances	Immunoglobulin M ; Antibodies, Viral
Language	English
Publishing date	2022-11-21
Publishing country	United States
Document type	Journal Article
ZDB-ID	390499-4
ISSN	1098-660X ; 0095-1137
ISSN (online)	1098-660X
ISSN	0095-1137
DOI	10.1128/jcm.01227-22
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Zs.A 1188: Show issues

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Article ; Online: Etiological analysis of discarded measles in the context of a measles outbreak among a highly immunized population.

Torner, Nuria / Mercader, Sara / Dominguez, Angela / Martinez, Ana / Costa, Josep / Sowers, Sun B / Abernathy, Emily S / Bellini, William J / Hickman, Carol J

Pediatrics international : official journal of the Japan Pediatric Society

2022 Volume 65, Issue 1, Page(s) e15430

Abstract: Background: Measles can lead to serious complications and remains an important cause of morbidity and mortality worldwide. In this study we aimed to assess the etiological diagnosis of discarded measles cases in the context of an outbreak among a highly ...

Abstract	Background: Measles can lead to serious complications and remains an important cause of morbidity and mortality worldwide. In this study we aimed to assess the etiological diagnosis of discarded measles cases in the context of an outbreak among a highly immunized population. Methods: We conducted a retrospective observational study of discarded measles cases from an outbreak that occurred from October 2006 to July 2007 in Catalonia. A confirmed case was defined as having a positive measles serum IgM result and/or a positive result by RT-PCR in urine and/or nasopharyngeal swab; or an epidemiological link to a confirmed case. Serum specimens were tested by a commercially available indirect-format and by an in-house capture-format measles IgM enzyme immunoassays. Results: Testing of 89 samples discarded for measles determined the etiologies for 10 (11.2%), including one rubella, three human herpes virus 6, and six measles infections. Of 381 confirmed cases in the outbreak, 10% had received at least one dose of the measles-mumps-rubella vaccine versus 54% of the discarded for measles (OR: 0.09; 95% CI: 0.06, 0.14; p < 0.001). Conclusions: Highly sensitive surveillance systems are critical to identifying cases, responding to outbreaks and verifying progress towards measles elimination. Molecular tools for measles detection and differential diagnosis, and collection of appropriate specimens for molecular and serological testing are essential to correctly diagnose suspected measles infection.
MeSH term(s)	Humans ; Measles/diagnosis ; Measles/epidemiology ; Measles/prevention & control ; Rubella/epidemiology ; Measles virus/genetics ; Disease Outbreaks ; Immunoglobulin M ; Antibodies, Viral
Chemical Substances	Immunoglobulin M ; Antibodies, Viral
Language	English
Publishing date	2022-11-28
Publishing country	Australia
Document type	Observational Study ; Journal Article
ZDB-ID	1470376-2
ISSN	1442-200X ; 1328-8067
ISSN (online)	1442-200X
ISSN	1328-8067
DOI	10.1111/ped.15430
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Zs.A 848: Show issues

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Article: Long-term Neutralizing Antibody Levels Against Measles and Rubella Viruses Among Adults With 3 Doses of Measles-Mumps-Rubella Vaccine.

Alonge, Oluwakemi D / Marin, Mona / Hickman, Carole J / Sowers, Sun B / Chen, Min-Hsin / Hao, Lijuan / Mercader, Sara / El-Badry, Elina / McClure, David L / Icenogle, Joseph P / Sugerman, David E / Crooke, Stephen N / Nguyen, Huong Q

Open forum infectious diseases

2024 Volume 11, Issue 1, Page(s) ofad700

Abstract: Background: A third dose of measles-mumps-rubella vaccine (MMR) may be administered for various reasons, but data on long-term immunity are limited. We assessed neutralizing antibody levels against measles and rubella among adults up to 11 years after ... ...

Abstract	Background: A third dose of measles-mumps-rubella vaccine (MMR) may be administered for various reasons, but data on long-term immunity are limited. We assessed neutralizing antibody levels against measles and rubella among adults up to 11 years after receipt of a third MMR dose. Methods: In this longitudinal study, healthy adults who received a third MMR dose as young adults (ages 18-28 years) were recalled around 5 years and 9-11 years after the third dose. Measles and rubella antibody levels were assessed by plaque-reduction and immunocolorimetric neutralization assays, respectively. Antibody concentrations <120 mIU/mL and <10 U/mL were considered potentially susceptible to measles and rubella, respectively. Geometric mean concentrations (GMCs) and 95% confidence intervals (CIs) over time were estimated from generalized estimating equation models. Results: Approximately 5 and 9-11 years after receipt of the third dose, 405 and 304 adults were assessed, respectively. Measles GMC was 428 mIU/mL (95% CI, 392-468 mIU/mL) 5 years postvaccination, declining to 381 mIU/mL (95% CI, 339-428 mIU/mL) 11 years postvaccination. At the last follow-up visit (9-11 years postvaccination), 10% of participants were potentially susceptible to measles infection. Rubella GMCs were stable throughout the follow-up period (63 U/mL to 65 U/mL); none of the participants was susceptible to rubella at the last follow-up visit. Conclusions: Eleven years after receiving a third MMR dose, measles and rubella neutralizing antibody levels remained high in adults. However, on the basis of waning antibody levels, some adults may become susceptible to measles infection over time despite receipt of 3 vaccine doses.
Language	English
Publishing date	2024-01-03
Publishing country	United States
Document type	Journal Article
ZDB-ID	2757767-3
ISSN	2328-8957
ISSN	2328-8957
DOI	10.1093/ofid/ofad700
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Article ; Online: Preservation of lymphocyte functional fitness in perinatally-infected and treated HIV+ pediatric patients displaying sub-optimal viral control.

Khanolkar, Aaruni / Muller, William J / Simpson, Bridget M / Cerullo, Jillian / Williams, Ruth / Sowers, Sun Bae / Matthews, Kiana / Mercader, Sara / Hickman, Carole J / D'Aquila, Richard T / Liu, Guorong

Communications medicine

2022 Volume 2

Abstract: Background: Host-pathogen dynamics associated with HIV infection are quite distinct in children versus adults. We interrogated the functional fitness of the lymphocyte responses in two cohorts of perinatally infected HIV+ pediatric subjects with early ... ...

Abstract	Background: Host-pathogen dynamics associated with HIV infection are quite distinct in children versus adults. We interrogated the functional fitness of the lymphocyte responses in two cohorts of perinatally infected HIV+ pediatric subjects with early anti-retroviral therapy (ART) initiation but divergent patterns of virologic control. We hypothesized that sub-optimal viral control would compromise immune functional fitness. Methods: The immune responses in the two HIV+ cohorts ( Results: We demonstrate that contrary to expectations pediatric HIV+ patients with sub-optimal viral control display no significant deficits in immune functional fitness. In fact, the patients that display better virologic control lack functional Gag-specific T cell responses and compared to healthy controls they display signaling deficits and an enrichment of mitogen-stimulated CD3 negative and positive lymphocyte clusters with suppressed cytokine production. Conclusions: These results highlight the immune resilience in HIV+ children on ART with sub-optimal viral control. With respect to HIV+ children on ART with better viral control, our data suggest that this cohort might potentially benefit from targeted interventions that might mitigate cell-mediated immune functional quiescence.
Language	English
Publishing date	2022-03-04
Publishing country	England
Document type	Journal Article
ISSN	2730-664X
ISSN (online)	2730-664X
DOI	10.1038/s43856-022-00085-9
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Article ; Online: Measles virus IgG avidity assay for use in classification of measles vaccine failure in measles elimination settings.

Mercader, Sara / Garcia, Philip / Bellini, William J

Clinical and vaccine immunology : CVI

2012 Volume 19, Issue 11, Page(s) 1810–1817

Abstract: In regions where endemic measles virus has been eliminated, diagnostic assays are needed to assist in correctly classifying measles cases irrespective of vaccination status. A measles IgG avidity assay was configured using a commercially available ... ...

Abstract	In regions where endemic measles virus has been eliminated, diagnostic assays are needed to assist in correctly classifying measles cases irrespective of vaccination status. A measles IgG avidity assay was configured using a commercially available measles-specific IgG enzyme immunoassay by modifying the protocol to include three 5-min washes with diethylamine (60 mM; pH 10.25) following serum incubation; serum was serially diluted, and the results were expressed as the end titer avidity index. Receiver operating characteristic analysis was used for evaluation and validation and to establish low (≤30%) and high (≥70%) end titer avidity thresholds. Analysis of 319 serum specimens expected to contain either high- or low-avidity antibodies according to clinical and epidemiological data indicated that the assay is highly accurate, with an area under the curve of 0.998 (95% confidence interval [CI], 0.978 to 1.000), sensitivity of 91.9% (95% CI, 83.2% to 97.0%), and specificity of 98.4% (95% CI, 91.6% to 100%). The assay is rapid (<2 h) and precise (standard deviation [SD], 4% to 7%). In 18 samples from an elimination setting outbreak, the assay identified 2 acute measles cases with low-avidity results; both were IgM-positive samples. Additionally, 11 patients (15 samples) with modified measles who were found to have high-avidity IgG results were classified as secondary vaccine failures; one sample with an intermediate-avidity result was not interpretable. In elimination settings, measles IgG avidity assays can complement existing diagnostic tools in confirming unvaccinated acute cases and, in conjunction with adequate clinical and epidemiologic investigation, aid in the classification of vaccine failure cases.
MeSH term(s)	Adult ; Antibodies, Viral/blood ; Antibody Affinity ; Disease Eradication ; Humans ; Immunoassay/methods ; Immunoglobulin G/blood ; Infant ; Measles/diagnosis ; Measles/immunology ; Measles/prevention & control ; Measles Vaccine/immunology ; Measles virus/immunology ; ROC Curve ; Sensitivity and Specificity ; Specimen Handling/methods ; Time Factors ; Treatment Failure
Chemical Substances	Antibodies, Viral ; Immunoglobulin G ; Measles Vaccine
Language	English
Publishing date	2012-09-12
Publishing country	United States
Document type	Evaluation Study ; Journal Article ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	2221082-9
ISSN	1556-679X ; 1556-6811
ISSN (online)	1556-679X
ISSN	1556-6811
DOI	10.1128/CVI.00406-12
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Zs.A 4000: Show issues

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Article ; Online: Development and Use of an Endpoint Titration Assay To Characterize Mumps IgG Avidity following Measles, Mumps, and Rubella Vaccination and Wild-Type Mumps Infection.

Mercader, Sara / McGrew, Marcia / Sowers, Sun B / Williams, Nobia J / Bellini, William J / Hickman, Carole J

mSphere

2018 Volume 3, Issue 5

Abstract: Waning mumps IgG antibody and incomplete IgG avidity maturation may increase susceptibility to mumps virus infection in some vaccinees. To measure mumps IgG avidity, serum specimens serially diluted to the endpoint were incubated on a commercial mumps- ... ...

Abstract	Waning mumps IgG antibody and incomplete IgG avidity maturation may increase susceptibility to mumps virus infection in some vaccinees. To measure mumps IgG avidity, serum specimens serially diluted to the endpoint were incubated on a commercial mumps-specific IgG enzyme immunoassay and treated with the protein denaturant diethylamine (60 mM, pH 10). End titer avidity indices (etAIs [percent ratio of detected diethylamine-resistant IgG at endpoint]) were calculated. Unpaired serum specimens (
MeSH term(s)	Antibodies, Viral/blood ; Antibody Affinity ; Case-Control Studies ; Female ; Humans ; Immunization Schedule ; Immunoglobulin G/blood ; Infant ; Male ; Measles-Mumps-Rubella Vaccine/administration & dosage ; Mumps/prevention & control ; Mumps virus/immunology ; United States
Chemical Substances	Antibodies, Viral ; Immunoglobulin G ; Measles-Mumps-Rubella Vaccine
Language	English
Publishing date	2018-09-12
Publishing country	United States
Document type	Journal Article
ISSN	2379-5042
ISSN (online)	2379-5042
DOI	10.1128/mSphere.00320-18
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Article ; Online: Decreased humoral immunity to mumps in young adults immunized with MMR vaccine in childhood.

Rasheed, Mohammed Ata Ur / Hickman, Carole J / McGrew, Marcia / Sowers, Sun Bae / Mercader, Sara / Hopkins, Amy / Grimes, Vickie / Yu, Tianwei / Wrammert, Jens / Mulligan, Mark J / Bellini, William J / Rota, Paul A / Orenstein, Walter A / Ahmed, Rafi / Edupuganti, Srilatha

Proceedings of the National Academy of Sciences of the United States of America

2019 Volume 116, Issue 38, Page(s) 19071–19076

Abstract: In the past decade, multiple mumps outbreaks have occurred in the United States, primarily in close-contact, high-density settings such as colleges, with a high attack rate among young adults, many of whom had the recommended 2 doses of mumps-measles- ... ...

Abstract	In the past decade, multiple mumps outbreaks have occurred in the United States, primarily in close-contact, high-density settings such as colleges, with a high attack rate among young adults, many of whom had the recommended 2 doses of mumps-measles-rubella (MMR) vaccine. Waning humoral immunity and the circulation of divergent wild-type mumps strains have been proposed as contributing factors to mumps resurgence. Blood samples from 71 healthy 18- to 23-year-old college students living in a non-outbreak area were assayed for antibodies and memory B cells (MBCs) to mumps, measles, and rubella. Seroprevalence rates of mumps, measles, and rubella determined by IgG enzyme-linked immunosorbent assay (ELISA) were 93, 93, and 100%, respectively. The index standard ratio indicated that the concentration of IgG was significantly lower for mumps than rubella. High IgG avidity to mumps Enders strain was detected in sera of 59/71 participants who had sufficient IgG levels. The frequency of circulating mumps-specific MBCs was 5 to 10 times lower than measles and rubella, and 10% of the participants had no detectable MBCs to mumps. Geometric mean neutralizing antibody titers (GMTs) by plaque reduction neutralization to the predominant circulating wild-type mumps strain (genotype G) were 6-fold lower than the GMTs against the Jeryl Lynn vaccine strain (genotype A). The majority of the participants (80%) received their second MMR vaccine ≥10 years prior to study participation. Additional efforts are needed to fully characterize B and T cell immune responses to mumps vaccine and to develop strategies to improve the quality and durability of vaccine-induced immunity.
MeSH term(s)	Adolescent ; Adult ; Antibodies, Neutralizing/blood ; Antibodies, Neutralizing/immunology ; Antibodies, Viral/blood ; Antibodies, Viral/immunology ; Child ; Child, Preschool ; Female ; Humans ; Immunity, Humoral/drug effects ; Immunity, Humoral/immunology ; Immunization ; Immunoglobulin G/blood ; Immunoglobulin G/immunology ; Infant ; Male ; Measles-Mumps-Rubella Vaccine/administration & dosage ; Measles-Mumps-Rubella Vaccine/pharmacology ; Mumps/immunology ; Mumps/prevention & control ; Mumps/virology ; Mumps virus/immunology ; Young Adult
Chemical Substances	Antibodies, Neutralizing ; Antibodies, Viral ; Immunoglobulin G ; Measles-Mumps-Rubella Vaccine
Language	English
Publishing date	2019-09-03
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	209104-5
ISSN	1091-6490 ; 0027-8424
ISSN (online)	1091-6490
ISSN	0027-8424
DOI	10.1073/pnas.1905570116
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Zs.A 1148: Show issues

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Article: Comparison of available methods to elute serum from dried blood spot samples for measles serology.

Mercader, Sara / Featherstone, David / Bellini, William J

Journal of virological methods

2006 Volume 137, Issue 1, Page(s) 140–149

Abstract: Six existing protocols for the extraction of serum from blood spots dried onto filter paper were compared. Assessment criteria included: detection of measles IgM and IgG by the Dade Behring Enzygnost immunoassays, volumes of recovered eluates, ... ...

Abstract	Six existing protocols for the extraction of serum from blood spots dried onto filter paper were compared. Assessment criteria included: detection of measles IgM and IgG by the Dade Behring Enzygnost immunoassays, volumes of recovered eluates, reproducibility, processing time and throughput, difficulty of protocol, equipment required, safety and estimated costs. Detection of measles IgM in eluates obtained by four of these protocols was as in serum, and significant differences were only observed in eluates from the two remaining protocols (p < 0.05). Significant differences were found between extraction protocols regarding measles-specific IgG detection when an IgG indeterminate DBS was analyzed (p < 0.05), but not when an IgG positive and negative DBS were studied. Sufficient eluate volumes were recovered for testing in the IgM Behring assay following all protocols but two. Sufficient eluate was recovered for testing in the IgG Behring assay following all six protocols. While all protocols were relatively easy to perform, only two protocols required less than 2h for completion. In general, compared protocols performed well on the extraction of antibodies from DBS for serology with differences being observed with eluate volume recovery, turn around time, required equipment and cost. An easy-to-implement protocol is proposed for the rapid extraction of serum for measles/rubella serology in outbreak situations for use in the World Health Organization Global Measles and Rubella Laboratory Network.
MeSH term(s)	Antibodies, Viral/blood ; Blood Specimen Collection ; Humans ; Immunoglobulin G/blood ; Immunoglobulin M/blood ; Measles/diagnosis ; Measles/immunology ; Measles virus/immunology ; Reagent Kits, Diagnostic ; Reproducibility of Results ; Sensitivity and Specificity ; Serologic Tests/methods ; Serum/immunology
Chemical Substances	Antibodies, Viral ; Immunoglobulin G ; Immunoglobulin M ; Reagent Kits, Diagnostic
Language	English
Publishing date	2006-10
Publishing country	Netherlands
Document type	Comparative Study ; Evaluation Studies ; Journal Article
ZDB-ID	8013-5
ISSN	1879-0984 ; 0166-0934
ISSN (online)	1879-0984
ISSN	0166-0934
DOI	10.1016/j.jviromet.2006.06.018
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Article ; Online: Dried blood spots on filter paper as an alternative specimen for measles diagnostics: detection of measles immunoglobulin M antibody by a commercial enzyme immunoassay.

Uzicanin, Amra / Lubega, Irene / Nanuynja, Miriam / Mercader, Sara / Rota, Paul / Bellini, William / Helfand, Rita

The Journal of infectious diseases

2011 Volume 204 Suppl 1, Page(s) S564–9

Abstract: Background: We compared the results of a serum-based measles immunoglobulin M (IgM) test with results of tests using paired reconstituted dried filter paper blood spot (DBS) samples to assess the feasibility of using DBS samples for measles diagnostic ... ...

Abstract	Background: We compared the results of a serum-based measles immunoglobulin M (IgM) test with results of tests using paired reconstituted dried filter paper blood spot (DBS) samples to assess the feasibility of using DBS samples for measles diagnostic procedures. Methods: We collected 588 paired serum and DBS samples from 349 children aged 8 months through 12 years at Mulago Hospital in Kampala, Uganda; of these samples, 513 (87%) were collected from children with a clinical diagnosis of measles 0-33 days after rash, and 75(13%) were collected from children hospitalized for other reasons. Eluted DBS and serum samples were tested using a commercial measles IgM enzyme immunoassay. Detection of viral RNA was attempted on a subset of 20 DBS by reverse-transcriptase polymerase chain reaction. Results: Among the 513 sample pairs collected from children with measles, the concordances for samples collected during days 0-6 and >1 week after rash were 95.7% and 100%, respectively (P<.01). The relative sensitivity and specificity of the DBS-based assay during the first week were 98.7% and 88.9%, respectively, and the sensitivity and specificity >1 week after rash were 100% and 100%, respectively. Viral RNA was detected in 5 (26%) of 19 DBS samples tested. Among 75 sample pairs collected from children hospitalized for other reasons, concordance was 94.7%. Conclusions: DBS samples are a feasible alternative sample for measles diagnostic procedures in high-incidence settings.
MeSH term(s)	Antibodies, Viral/blood ; Case-Control Studies ; Child ; Child, Preschool ; Female ; Humans ; Immunoenzyme Techniques/methods ; Immunoglobulin M/blood ; Infant ; Male ; Measles/blood ; Measles/diagnosis ; Measles/epidemiology ; Paper ; Specimen Handling/methods ; Uganda/epidemiology
Chemical Substances	Antibodies, Viral ; Immunoglobulin M
Language	English
Publishing date	2011-07
Publishing country	United States
Document type	Comparative Study ; Journal Article ; Research Support, U.S. Gov't, P.H.S.
ZDB-ID	3019-3
ISSN	1537-6613 ; 0022-1899
ISSN (online)	1537-6613
ISSN	0022-1899
DOI	10.1093/infdis/jir088
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