Mumps Virus-Specific Antibody Titers from Pre-Vaccine Era Sera: Comparison of the Plaque Reduction Neutralization Assay and Enzyme Immunoassays

Department of Pediatrics, Johns Hopkins University, Baltimore, Maryland, United States
Journal of Clinical Microbiology (Impact Factor: 3.99). 10/2005; 43(9):4847-51. DOI: 10.1128/JCM.43.9.4847-4851.2005
Source: PubMed


Mumps virus-neutralizing antibodies are believed to be the most predictable surrogate marker of protective immunity. However, assays used to detect neutralizing antibodies, such as the plaque reduction neutralization (PRN) assay, are labor- and time-intensive and consequently are often supplanted by the more rapid and inexpensive enzyme immunoassay (EIA) technique. For virus infections for which international antibody standards exist and are bridged to clinical studies of protection (e.g., measles and rubella), the EIA has been successfully used to determine immune surrogate endpoints, yet no such international reference exists for mumps serology. Since both virus-neutralizing and nonneutralizing antibodies are measured in the EIA, in the absence of a mumps serological standard, the EIA may be prone to yielding false-positive results when utilized for assessing surrogate markers of protective immunity. Moreover, since mumps virus-specific antibody titers are generally low in comparison to antibody levels induced by other viruses and EIA procedures often employ relatively high serum dilution factors, the EIA may be prone to yielding false-negative results. To examine these issues, a PRN assay and two commercially available EIA kits were used to evaluate wild-type mumps virus serological responses in human serum samples from the pre-mumps vaccine era. Our results indicate that the PRN assay is a more sensitive and specific method of measuring serological responses to wild-type mumps virus.

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Available from: Jeremy Mauldin, Apr 24, 2014
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    • "While an elevated titer may not necessarily provide a consistent diagnostic marker even within the context of a known outbreak, it is possible that moderate boosting of IgG in the acute blood may interfere with the ability to detect a fourfold rise between acute and convalescent serum samples. The IgG response as measured by the EIA assay does not distinguish between neutralizing and non-neutralizing antibodies, so that a change in neutralizing titer may be masked by high concentrations of non-neutralizing antibodies [Mauldin et al., 2005]. In addition, the time course and magnitude of the immune response may be altered in persons who have received one or more doses of MMR. "
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    ABSTRACT: Following the clinical diagnosis of the first case of mumps on September 22, 2006 at the University of Virginia (UVA), 52 suspected cases were identified through active surveillance for mumps by the end of December 2006. Samples were collected from 47 students who presented with parotitis despite a documented history of two doses of measles, mumps, and rubella (MMR) vaccine. Six of 47 serum samples (13%) were positive for mumps IgM, and 46/47 specimens were positive for mumps IgG. Endpoint titration of acute phase serum samples from laboratory-confirmed cases did not provide evidence that elevated serum IgG is a consistent marker for infection among cases due to secondary vaccine failure. Buccal swab samples from 39 of the 47 students were tested by real-time reverse transcription-polymerase chain reaction (RT-PCR) and/or viral culture. Mumps virus or mumps RNA was detected in 12 of 39 buccal samples (31%). Genetic analysis of the virus from the outbreak at UVA indicated that the outbreak was not linked to the large mumps outbreak in the Midwestern US that occurred earlier in 2006. Our findings support the use of viral detection to improve laboratory diagnosis of mumps among persons who have received two doses of MMR.
    Journal of Medical Virology 10/2009; 81(10):1819-25. DOI:10.1002/jmv.21557 · 2.35 Impact Factor
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    • "These observations have raised public health concerns and suggested that further detailed studies on mumps vaccine efficacy and population susceptibility are warranted. Plaque reduction neutralization test (PRNT) measures the functional antibody (of any class) by in vitro virus neutralization and is considered the " gold-standard " assay for assessing serological correlates of protection (Mauldin et al., 2005). However, PRNT is technically demanding, not easy to automate, and has limitations for screening the large numbers of sera needed for epidemiological investigations. "
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    ABSTRACT: Although the plaque reduction neutralization test (PRNT) is considered the "gold-standard" assay for measuring neutralizing antibodies for mumps, it is technically demanding, slow and requires large serum volumes, which limits its use for investigating mumps vaccine efficacy and population susceptibility. Therefore, an immunocolourimetric-based focus reduction neutralization test (FRNT) was developed and validated against PRNT using 30 blood donor plasma samples (16 positive, 5 equivocal, and 9 negative for mumps IgG by EIA). The samples were tested in triplicate by FRNT and PRNT in 10 and 4 separate assay runs, respectively, and 50% neutralizing antibody titres calculated using the Kärber formula. There was good correlation between the two neutralization assays (R(2)=0.88). Inter-assay variation for FRNT titres was 2-fold, compared to a 3-fold variation for PRNT titres. From the distribution of results, a positive cut-off for FRNT was defined as 1:4. In conclusion, FRNT has similar sensitivity to the PRNT and offers the advantage of speed (2 days vs. 7 days), reduced sample volume (40 microL vs. 150 microL), and the possibility of automation using 96-well plates. FRNT appears to be a good substitute for PRNT for characterising the immune response to mumps and for vaccine efficacy studies.
    Journal of virological methods 09/2009; 163(1):153-6. DOI:10.1016/j.jviromet.2009.09.006 · 1.78 Impact Factor
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    • "With regards to the sensitivity of the assays used, it should be taken into account that false negative ELISA results might distort the actual rate of humoral immunity [12]. These assays are nevertheless commonly used as screening tests to determine immunity status because they are less labor- and time-intensive than cell culture-based neutralisation assays which detect neutralising antibodies. "
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    ABSTRACT: Medical students come into contact with infectious diseases early on their career. Immunity against vaccine-preventable diseases is therefore vital for both medical students and the patients with whom they come into contact. The purpose of this study was to compare the medical history and serological status of selected vaccine-preventable diseases of medical students in Germany. The overall correlation between self-reported medical history statements and serological findings among the 150 students studied was 86.7 %, 66.7 %, 78 % and 93.3 % for measles, mumps, rubella and varicella, conditional on sufficient immunity being achieved after one vaccination. Although 81.2 % of the students' medical history data correlated with serological findings, significant gaps in immunity were found. Our findings indicate that medical history alone is not a reliable screening tool for immunity against the vaccine-preventable diseases studied.
    BMC Public Health 02/2008; 8(1):121. DOI:10.1186/1471-2458-8-121 · 2.26 Impact Factor
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