Development of a new neutralization test for measles virus

Fujino, Motoko; Yoshida, Naoko; Kimura, Keiko; Zhou, Jian; Motegi, Yoshie; Komase, Katsuhiro; Nakayama, Tetsuo

doi:10.1016/j.jviromet.2007.01.001

Cited by 20 publications

(20 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Fujino et al [19] reported the neutralization test for measles virus using a GFP-expressing recombinant measles virus to evaluate the neutralizing antibody titer by Fluorescent EIA reader. Here, a recombinant mumps Hoshino vaccine strain expressing GFP was developed to check the expression of GFP instead of observing the appearance of CPE or plaque counting.…”

Section: Discussionmentioning

confidence: 99%

“…The plates were incubated for 7 days. In order to calculate the titers automatically, the plates were processed to detect fluorescence intensity (Fluoro-Units: FU) at an emission wavelength of 528 nm and excitation wavelength of 485 nm using a fluorescence reader, FLx800 (Bio-Tek Instruments, Vermont, USA), similar to a method used to detect measles neutralizing antibodies [19]. To evaluate the requirement of complement, various concentrations of guinea pig complement (Denka Seiken, Tokyo, Japan) were added to the neutralization mixture of serially diluted serum with challenge virus.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

A New Method for the Detection of Neutralizing Antibodies against Mumps Virus

et al. 2013

Self Cite

View full text Add to dashboard Cite

Neutralization test is the most reliable method of evaluating immunity against viral diseases but there is no standard procedure for mumps virus, with tests differing in the infectivity of the challenge virus, 50% plaque reduction or complete inhibition of cytopathic effects (CPE), and usage of complement. A reliable, easy, and simple neutralization test for mumps virus was developed in this study. A recombinant mumps virus expressing GFP was generated as a challenge virus. Complement was added to the neutralizing mixture at 1∶200 when stocked serum samples were used. Neutralizing antibody titers were expressed as the reciprocal of the highest dilution that did not exceed two-fold of FU values (GFP expression) of the cell control wells. A total of 1,452 serum samples were assayed by inhibition of GFP expression in comparison with those examined by conventional 100% inhibition of CPE. 1,367 (94.1%) showed similar neutralizing antibody titers when examined by both methods. The GFP expression inhibition assay, using a recombinant mumps virus expressing GFP, is a simple and time- saving method.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

A New Method for the Detection of Neutralizing Antibodies against Mumps Virus

et al. 2013

Self Cite

View full text Add to dashboard Cite

show abstract

“…Vervet monkeys have often been used to provide red blood cells for the measles hemagglutination inhibition test. However, the number of captive vervet monkeys is decreasing in Japan because of new diagnostic methods for measles (Fujino et al, 2007). Although there is a US primate center with a vervet monkey colony (http://www.ncrr.nih.gov/comparative_medicine/resource_directory/primates.asp#verv), funds are insufficient for maintaining such a huge facility in Japan.…”

Section: Discussionmentioning

confidence: 99%

Transthyretin Amyloidosis in Aged Vervet Monkeys, as a Candidate for the Spontaneous Animal Model of Senile Systemic Amyloidosis

Nakamura¹,

Ueda²,

Ageyama³

et al. 2011

Amyloidosis - Mechanisms and Prospects for Therapy

View full text Add to dashboard Cite

“…Recombinant viruses expressing various reporter genes, like GFP, luciferase, and ␤-galactosidase genes, provide us with excellent tools to aid in the assessment of protective neutralizing antibodies (14). For MV, alternatives to the classical PRN assay include H protein-based EIA (3), neutralization EIA, (13) and a neutralization test, based on a recombinant GFP-expressing MV (10). The last study applied a different strain of MV (AIK-C) and a longer and more complicated assay protocol, including a semisolid agarose overlay.…”

Section: Discussionmentioning

confidence: 99%

Development of a Novel Efficient Fluorescence-Based Plaque Reduction Microneutralization Assay for Measles Virus Immunity

Haralambieva

Ovsyannikova

Vierkant

et al. 2008

Clin Vaccine Immunol

View full text Add to dashboard Cite

The measurement of functional measles virus-specific neutralizing antibodies is of considerable interest for vaccine-related research. In this study, we developed and standardized a simple, rapid, highly sensitive, and reproducible fluorescence-based plaque reduction microneutralization (PRMN) assay with visual and automated readout, using a recombinant measles virus engineered to express enhanced green fluorescent protein.The assay is performed in micro format, requires less time to complete (2 versus 4 to 7 days), and is less labor-intensive and less costly than the classical plaque reduction neutralization (PRN) test, widely accepted as the "gold standard" in measles serology. Two available WHO international anti-measles virus standards and one in-house reference serum were used to develop and standardize the new assay. The mean PRMN values from repeated assays were found to be similar to those reported in the literature or assigned to the WHO standards by the classical PRN assay. For validation, we used three groups of low, moderate, and high measles virus vaccine responders' sera with moderate values of correlation in antibody levels (mIU/ml) between PRMN and the Dade Behring immunoglobulin G enzyme immunoassay (EIA). The PRMN assay was more sensitive at low antibody levels and more informative in terms of protection than this commercial EIA. In conclusion, we have developed and validated a sensitive and high-throughput measles virus-specific PRMN that can be readily used in large population-based measles studies.Measles is a highly communicable infectious disease, and it remains the leading cause of vaccine-preventable childhood mortality in developing countries and is still a major public health concern in developed countries (15). Measles outbreaks are known to occur even in highly vaccinated populations despite the availability of an effective live attenuated measles virus (MV) vaccine (17).Neutralizing and protective antibodies are directed against the two surface MV glycoproteins, the hemagglutinin (H) and fusion (F) proteins, and are sufficient to provide protection (4). Therefore, diagnostic measures of sufficient levels of functional neutralizing MV-specific antibodies correlate with protection. Neutralizing antibodies are operationally defined by a standard plaque reduction neutralization (PRN) test as antibodies that prevent a cytopathic effect and formation of plaques, using a laboratory MV strain (Edmonston) on Vero cells (4). The PRN assay, an enhanced version of the neutralization test, is widely accepted as the "gold standard" in MV serology and measures the serum dilution capable of preventing 50% of plaque formation by MV (PRN titer; 50% neutralizing dose [ND 50 ]) (1). However, the classical PRN assays are slow (4 to 7 days), labor-intensive, and require large volumes of test components (since they are conducted in 24-well and 12-well plates using semisolid cell overlay and staining procedures) and are impractical for large numbers of samples. The availability of a relatively simple, more...

show abstract

Development of a new neutralization test for measles virus

Cited by 20 publications

References 15 publications

A New Method for the Detection of Neutralizing Antibodies against Mumps Virus

A New Method for the Detection of Neutralizing Antibodies against Mumps Virus

Transthyretin Amyloidosis in Aged Vervet Monkeys, as a Candidate for the Spontaneous Animal Model of Senile Systemic Amyloidosis

Development of a Novel Efficient Fluorescence-Based Plaque Reduction Microneutralization Assay for Measles Virus Immunity

Contact Info

Product

Resources

About