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Comparison of five methods for the detection of measles specific immunoglobulin G antibody
Abstract
In this work we compare the sensitivity, specificity and predictive values of hemagglutination inhibition (HI), immunofluorescent assay (IFA), biotin-streptavidin immunofluorescent assay (B/SA-IFA), enzyme-linked immunosorbent assay (EIA) and plaque neutralization test (PN). This study includes serum samples from children taken before and after vaccination, children with clinically diagnosed measles and household contacts. EIA were the most specific and better serological diagnostic test. HI and IFA had high sensitivity but low specificity. An alternative to EIA is B/SAIFA, which is cheaper than EIA and can be used in the study of small outbreaks or in isolated cases.
... Traditionally, various neutralization assays have been developed for many different viruses. Plaque reduction assays have been widely used to evaluate the neutralizing antibody responses against viruses that can form plaques in infected cells, such as rubella (Rhim and Schell, 1967;Sato et al., 1979), flavivirus (Russell and Nisalak, 1967;Ibrahim et al., 1968), vaccinia (Kitamura et al., 1973;Newman et al., 2003) and measles viruses (Orenstein et al., 1987;Vazquez-Rosales et al., 1996). However, unpublished data including our own have observed that the plaques formed in SCV infected target cells (such as Vero E6) were poor and could not be visualized clearly unless a microscope is used. ...
... Our study demonstrated that an improved plaque reduction assay has been established to measure the neutralizing antibody activities against SCV infection. Previously plaque assays have been used successfully for a wide range of viral infection and neutralization studies including vaccinia (Kitamura et al., 1973;Newman et al., 2003), measles (Orenstein et al., 1987;Vazquez-Rosales et al., 1996) and rabies (Yoshino and Morishima, 1971;Wiktor and Clark, 1973;Cardoso and Pilz, 2004). However, the neutralizing antibody titers observed by the plaque reduction assay in our study were lower than those determined by the CPE assay or the neutral red staining assay. ...
Accurate assessment of neutralizing antibody activities is important either for patients infected with Severe Acute Respiratory Syndrome (SARS) or for animals and volunteers immunized with the experimental vaccines against the SARS associated coronavirus (SCV). However, the current assay based on the cytopathic effect (CPE) which has been frequently cited in literature has several limitations. The CPE assay relies on the visual observation on the damage of SCV infected target cells under a microscope. It is subjected to observer variations and it is difficult to generate a quantitative determination of neutralizing activities based on the level of CPE. In the current study, we established the utility of two additional assays to measure the neutralizing activities against SCV: the plaque reduction (PR) and the neutral red staining (NRS) assays. The PR assay described in this study was modified from the traditional viral plaque reduction assay by using an improved crystal staining method to achieve better plague formation in SCV infected Vero E6 cells. The NRS neutralization assay was adopted from a similar system used for detecting neutralizing antibody responses against human immunodeficiency virus type 1 (HIV-1). In this assay, the protective effect of neutralizing antibodies was determined by the cell viability which is measured by the uptake of neutral red dye at A540. The neutralizing antibody titers can be easily determined with either of the two new assays. In this report, we described the utility of these two new neutralization assays in measuring the neutralizing activities against SCV infection from rabbit sera immunized with various forms of spike protein of SCV.
Aim:
Measles during pregnancy has deleterious effects on both perinatal and maternal outcomes. In Japan, local epidemics of measles and cases of measles during pregnancy are still being reported; therefore, the seroprevalence of antibodies to measles is suspected to be still not sufficient. The aim of this study was to analyze the seroprevalence of antibodies to measles in Japanese pregnant women and estimate the percentage of these women who require vaccination or revaccination against measles.
Material and methods:
We analyzed the seroprevalence of immunity to measles by the neutralization test in 10 349 pregnant women in the first trimester managed at the National Center for Child Health and Development between February 2004 and December 2010. The neutralization test titers were interpreted as follows: ≧1:8, seropositive; =4, low-positive; ≦4, seronegative.
Results:
Of the total number of pregnant women tested, 7408 (71.6%) were seropositive, 1864 (18.0%) were low-positive, and 1079 (10.4%) were seronegative for measles antibodies, respectively.
Conclusion:
Our results revealed that 28% of our pregnant population was seronegative or low-positive for measles antibodies, and thought to require revaccination or vaccination. Screening for measles immunity might be advisable for women of childbearing age.
Measles continues to be a major disease of both human and nonhuman primates. The dot immunobinding assay, a modified enzyme immunoassay, permits the detection of measles virus antibody in the nonlaboratory setting with either serum or whole blood collected on filter paper.
A biotin-avidin-amplified indirect immunofluorescence method was used to demonstrate specific serum immunoglobulin M (IgM) antibodies in nephropathia epidemica, the Scandinavian type of hemorrhagic fever with renal syndrome. The antigen in the test was the cross-reacting agent of Korean hemorrhagic fever, Hantaan virus. Sixty-two serum samples from 15 patients with clinically typical nephropathia epidemica were analyzed. Eleven patients had specific IgM in one or more serum samples. The IgM could be demonstrated from day 2 up to day 37, and all patients had detectable specific IgM within 15 days after the onset of disease. In 49 control serum samples, no specific IgM could be detected, indicating a high specificity for the method. The findings demonstrate that the biotin-avidin-amplified immunofluorescence IgM assay is a useful tool in the diagnosis of early nephropathia epidemica disease.
A convenient and flexible enzyme-linked immunosorbent assay system for the detection of specific antibodies to measles virus
has been developed. In this system infected cells are desiccated on 96-well microtiter plates and stored at room temperature.
After incubation of samples to be tested in the cell-coated plates and subsequent washing, bound antibodies are detected with
a peroxidase-conjugated staphylococcal protein A probe. After another washing and the addition of the appropriate substrate,
the amount of bound probe is estimated by colorimetric analysis. This technique offers several advantages. The need for a
purified viral antigen source is obviated. The plates are easily prepared and can be stored for months at room temperature.
Major viral epitopes, including surface glycoproteins as well as cytoplasmic viral antigens, are preserved despite desiccation.
The method is more sensitive than the conventional means of virus-specific antibody detection.
Epidemiological studies of measles and measles immunization frequently require determination of measles antibody status. In developing countries, where venipuncture is frequently unacceptable and where refrigerated storage of serum specimens is often unavailable, microtiter techniques not requiring refrigeration are required. We developed a filter paper technique that measures measles hemagglutination inhibition antibody and meets these criteria. Comparison of separately collected venous blood and peripheral blood collected on filter paper demonstrated 97% agreement in terms of presence or absence of antibody. In 30 of 32 measles specimens, 94% of titers were the same or varied by less than 2 twofold dilutions.
An amplification of the immunofluorescence technique which uses biotinylated antibody and fluoresceinated avidin is described. By introducing a sandwich technique using fluorescein-conjugated goat anti-avidin, a 5-fold enhancement of staining over the conventional immunofluorescence method is achieved, and the brightness is more than twice that achieved with the simple biotin-fluoresceinated avidin reaction.