Towards elimination: measles susceptibility in Australia and 17 European countries.

OBJECTIVE: To evaluate age-specific measles susceptibility in Australia and 17 European countries. METHODS: As part of the European Sero-Epidemiology Network 2 (ESEN2), 18 countries collected large national serum banks between 1996 and 2004. These banks were tested for measles IgG and the results converted to a common unitage to enable valid intercountry comparisons. Historical vaccination and disease incidence data were also collected. Age-stratified population susceptibility levels were compared to WHO European Region targets for measles elimination of < 15% in those aged 2-4 years, < 10% in 5-9-year-olds and < 5% in older age groups. FINDINGS: Seven countries (Czech Republic, Hungary, Luxembourg, Spain, Slovakia, Slovenia and Sweden) met or came very close to the elimination targets. Four countries (Australia, Israel, Lithuania and Malta) had susceptibility levels above WHO targets in some older age groups indicating possible gaps in protection. Seven countries (Belgium, Bulgaria, Cyprus, England and Wales, Ireland, Latvia and Romania) were deemed to be at risk of epidemics as a result of high susceptibility in children and also, in some cases, adults. CONCLUSION: Although all countries now implement a two-dose measles vaccination schedule, if the WHO European Region target of measles elimination by 2010 is to be achieved higher routine coverage as well as vaccination campaigns in some older age cohorts are needed in some countries. Without these improvements, continued measles transmission and outbreaks are expected in Europe.

Comparison of rubella seroepidemiology in 17 countries: progress towards international disease control targets.

OBJECTIVE: To standardize serological surveillance to compare rubella susceptibility in Australia and 16 European countries, and measure progress towards international disease-control targets. METHODS: Between 1996 and 2004, representative serum banks were established in 17 countries by collecting residual sera or community sampling. Serum banks were tested in each country and assay results were standardized. With a questionnaire, we collected information on current and past rubella vaccination programmes in each country. The percentage of seronegative (< 4 IU/ml) children (2-14 years of age) was used to evaluate rubella susceptibility, and countries were classified by seronegativity as group I (< 5%), group II (5-10%) or group III (> 10%). The proportion of women of childbearing age without rubella protection (< or = 10 IU/ml) was calculated and compared with WHO targets of < 5%. FINDINGS: Only Romania had no rubella immunization programme at the time of the survey; the remaining countries had a two-dose childhood schedule using the measles, mumps and rubella (MMR) vaccine. The percentage of susceptible children defined five countries as group I, seven as group II and four as group III. Women of childbearing age without rubella protection were < 5% in only five countries. CONCLUSION: Despite the low reported incidence in many countries, strengthening the coverage of the routine two-dose of MMR vaccine among children is needed, especially in group III countries. Catch-up campaigns in older age groups and selective targeting of older females are needed in many countries to ensure necessary levels of protective immunity among women of childbearing age.

Vaccinated students with negative enzyme immunoassay results show positive measles virus-specific antibody levels by immunofluorescence and plaque neutralisation tests.

BACKGROUND: The prevalence of measles antibodies was investigated by an enzyme immunoassay (EIA) in students aged 14 every year since 1996 in a Swiss municipality. This region has wide measles vaccine coverage (first dose > or = 95%, second dose > or = 65%) without any reported measles outbreaks since 20 years. In 2003 and 2004, in contrast to previous years, surprisingly many negative results (33% and 54%, respectively) were observed. OBJECTIVES: To corroborate the measles antibody values by different methods. STUDY DESIGN: Serum samples from 101 students with known vaccination status were available. Sera with equivocal and negative results obtained by two different EIAs were retested by indirect immunofluorescence test (IFT) and plaque neutralisation test (PNT). RESULTS: Retesting by IFT showed a positive result in 17/21 sera (81%) and retesting by PNT indicated that 46/49 sera (94%) were positive; the three sera with negative PNT result were from unvaccinated individuals. Only 3/96 vaccinated students showed measles antibodies below the putative protective level of 0.2 IU/ml after retesting by PNT. CONCLUSIONS: Negative EIA results should be interpreted with caution in a widely vaccinated population without booster by circulation of wild viruses. Retesting by IFT or PNT is recommended.

European seroepidemiology network 2: Standardisation of assays for seroepidemiology of varicella zoster virus.

BACKGROUND: The aim of the European Sero-Epidemiology Network (ESEN2) is to harmonise the serological surveillance of vaccine-preventable diseases in Europe. OBJECTIVE: To allow comparison of antibody prevalence in different countries by standardising results into common units. STUDY DESIGN: For varicella zoster virus (VZV), a reference laboratory established a panel of 148 samples, characterised by indirect enzyme-immunoassay (ELISA), indirect immunofluorescence, and complement fixation test. Fifty-seven samples were also studied by the fluorescence antibody to membrane antigen test. The geometric mean of the antibody activity (GMAA) obtained from four ELISA determinations was used to characterise each sample of the panel as positive (GMAA: >100 mIU/ml), equivocal (GMAA: 50-100 mIU/ml) or negative (GMAA: <50 mIU/ml) for antibody to VZV (anti-VZV). Thirteen laboratories, using five different ELISA tests, tested the panel. RESULTS: Agreement with the reference laboratory was above 85% in all cases, and the R(2) values obtained from regression analysis of the quantitative results were always higher than 0.87. Finally, the regression equations could be used to convert national values into a common unitage. CONCLUSION: This study confirmed that results for anti-VZV obtained by different ELISA methods can be converted into common units, enabling the comparison of the seroprevalence profiles obtained in the participant countries.

Open randomized trial comparing the immunogenicity and safety of a new measles-mumps-rubella vaccine and a licensed vaccine in 12- to 24-month-old children.

BACKGROUND: A trivalent measles-mumps-rubella (MMR) vaccine (MMR Berna) has been developed with a new mumps component, BBM-18, to replace a previously licensed MMR vaccine containing the Rubini mumps strain. Previous studies showed Rubini to confer insufficient long term protection against mumps infection. This study compared the immunogenicity and safety of MMR Berna, which is produced entirely in human diploid cells, with those of the licensed vaccine M-M-RVax (Merck & Co.). METHODS: We vaccinated 467 subjects, 12-24 months of age, in an open, randomized (1:1), phase II, multicenter study. Antibody titers were determined for each vaccine component with a plaque neutralization test (PNT) and a commercial enzyme-linked immunosorbent assay. Solicited local and systemic reactions were recorded in subject diaries for 6 weeks after vaccination. RESULTS: Seroconversion rates 6 to 8 weeks after vaccination for measles and rubella were statistically comparable for the 2 vaccines. However, mumps seroconversion rates were highly assay dependent, with significant differences being measured with the enzyme-linked immunosorbent assay (Berna, 77.4%; Merck, 91.3%; P < 0.001) but not the PNT (Berna, 84.8%; Merck, 87.6%; P = 0.42). The overall rate of systemic reactions was lower in the MMR Berna group (36.8% versus 45.9%; P < 0.05), including a significantly lower rate of fever of >38 degrees C (37.2% versus 51.8%; P < 0.01). CONCLUSIONS: MMR Berna was statistically noninferior to M-M-RVax with respect to seroconversion rates, and the BBM-18 strain elicited a level of functional antimumps antibodies comparable to the Jeryl Lynn strain, as measured with the PNT. Overall, MMR Berna was better tolerated than the comparison vaccine, particularly with respect to the frequency of fever.

Serosurvey of measles, mumps and rubella antibodies in Saudi children.

OBJECTIVE: A serosurvey study to evaluate the proportion of children with antibodies against diseases targeted by the Expanded Program of Immunization in the Kingdom of Saudi Arabia. METHODS: Using multistage sampling techniques, we collected samples and sent them for laboratory assay from the following age groups; 100 samples at 6 months, 12 months, 18 months, 6 years, 13 years, and 17 years. We conducted the study from September 2001 to February 2002. We assayed sera for measles, rubella, and mumps antibodies in the measles-mumps-rubella reference laboratory in Germany, using enzyme immunoassay and plaque neutralization (PN) as a backup test for equivocal and negative samples. We only carried out a backup test for measles samples. RESULTS: The age group of 6 months had the highest proportion with negative measles antibodies. After adding the backup test (PN), the proportions of children with protective measles antibody were; 64% at 6 months, 87% at 12 months, 91% at 18 months, 75% at 6 years, 96% at 13 years, and 98% at 17 years. Rubella antibody positivity rates (>7 IU) were 28% at 6 months, 49% at 12 months, 97% at 18 months, 98% at 6 years, and 100% at 13 years. While positivity rates in mumps were 14% at 6 months, 29% at 12 months, 59% at 18 months, 64% at 6 years, and 75% at 13 years. CONCLUSION: The unexpected low proportion of children with protective level at 6 years, despite being vaccinated with 2 measle doses is an important phenomenon. This reflects the interference between the first and the second measles dose. The Ministry of Health decided to conduct a catch up campaign targeting 1st through 3rd grade primary schools, who did not catch the mass campaign conducted in 2000. Also, this supports the decision taken by the ministry to change the measles immunization schedule to MMR at 12 months and a second dose at 6 years of age.

Effectiveness of measles vaccination after household exposure during a measles outbreak: a household contact study in Coburg, Bavaria.

BACKGROUND: A measles outbreak was recently observed in Coburg, Bavaria, in a population with vaccination rates of 76.5% in 5- to 6-year-old children in the years preceding the outbreak. Only a small proportion of children had received 2 vaccinations against measles. Vaccine effectiveness is estimated in a household contact study and also by a screening method. METHODS: A household contact study was conducted in families with at least 1 measles case by standardized computer-assisted telephone interviews to assess secondary attack rate and to estimate vaccine effectiveness. Vaccine effectiveness was also estimated with Farrington's screening method with information from school entry examinations and from questionnaires of confirmed measles cases in the Coburg outbreak. RESULTS: Thirty-eight children were primary cases. Of their contacts, 20 children were included in the study as secondary cases (1 vaccinated), and 23 children were contacts who did not develop measles (12 vaccinated once and 4 vaccinated twice), resulting in a vaccine effectiveness of 90% (95% confidence interval, 35-97%) for one vaccine dose. The proportion of the population vaccinated reached 81.5% during the outbreak and the proportion of the cases vaccinated was 10.9%, resulting in a vaccine effectiveness estimated using the screening method of 97.2% (95% confidence interval, 95.7-98.3%). CONCLUSIONS: With the use of 2 approaches to estimate the effectiveness of measles vaccination, a consistently high vaccine effectiveness of 90% or above was shown during a measles outbreak in Western Europe.

Probing neutralizing-antibody responses against emerging measles viruses (MVs): immune selection of MV by H protein-specific antibodies?

Measles virus (MV) infection and vaccination induce long-lasting immunity and neutralizing-antibody responses that are directed against the MV haemagglutinin (H) and the fusion (F) protein. A new MV genotype, D7, emerged recently in western Germany and rapidly replaced the long-term endemically circulating genotypes C2 and D6. Analysis of the H gene of C2, D6, D7 and vaccine viruses revealed uniform sequences for each genotype. Interestingly, a consistent exchange of seven distinct amino acids in the D7 H was observed when compared with residues shared between C2, D6 and vaccine viruses, and one exchange (D416-->N) in the D7 H was associated with an additional N-linked glycosylation. In contrast, the F gene is highly conserved between MVs of these genotypes. To test whether the D7 H protein escapes from antibody responses that were raised against earlier circulating or vaccine viruses, the neutralizing capacity of mAbs recognizing seven distinct domains on the H of an Edmonston-related MV was compared. The mAbs revealed a selective and complete loss of two neutralizing epitopes on the D7 H when compared with C2, D6 and vaccine viruses. To assess whether these alterations of the D7 H affect the neutralizing capacity of polyclonal B-cell responses, genotype-specific antisera were produced in cotton rats. However, no significant genotype-dependent difference was found. Likewise, human sera obtained from vaccinees (n=7) and convalescents (n=6) did not distinguish between the MV genotypes. Although the hypothesis of selection of D7 viruses by pre-existing neutralizing antibodies is compatible with the differing pattern of neutralizing epitopes on the H protein, it was not confirmed by the results of MV neutralization with polyclonal sera.

Laboratory investigations are indispensable to monitor the progress of measles elimination--results of the German Measles Sentinel 1999-2003.

BACKGROUND: The elimination of measles is a goal set by the World Health Organisation to be reached by 2010 in the European region. OBJECTIVES: To enhance the measles surveillance in Germany, a country-wide laboratory supported a sentinel was established. STUDY DESIGN: A network of >1200 representatively distributed practitioners reported detailed data on all clinically diagnosed cases and provided specimens for laboratory diagnosis. RESULTS: A total of 3225 suspected cases were reported between October 1999 and December 2003. The incidence in Western Germany decreased from >15 cases per 100,000 population to one case in 2003, while in Eastern Germany <1 case per 100,000 population was observed during these years. Laboratory investigations were undertaken in 40% of cases in 2000/2001. This rate increased to 79% in 2003. Simultaneously, the rate of confirmed cases dropped from 60% in the former years to 23% in 2003. Measles virus (MV) detection by serology and by PCR revealed concordant results in 92%. Most suspected cases (85%) were unvaccinated with 66% being laboratory confirmed. Only 10% of suspected cases occurred in vaccinated individuals and very few (22%) could be confirmed. Analyses of confirmed measles in vaccinated patients (n = 49) revealed 24.5% primary vaccine failures, 24.5% reinfections after successful vaccination and 31% MV infection before or shortly after vaccination. The genetic characterisation of 389 MV isolates identified eight genotypes: B3, C2, D4, D5, D6, D7, G2 and H1. Only the C2, D6 and D7 MV genotypes circulated endemically in Western Germany. The newly emerged MV D7 almost completely replaced the pre-existing C2 and D6 MVs in 2001. The few measles cases detected in Eastern Germany were mostly caused by imported MVs. CONCLUSION: The data demonstrate that laboratory investigations including molecular methods are an indispensable tool for surveillance in all countries advanced in measles elimination.

Passive acquired immunity against measles in infants born to naturally infected and vaccinated mothers.

BACKGROUND: Currently, the majority of newborns in Poland are born to mothers who have been immunized against measles. The aim of this study was to compare the maternal measles antibody titers of infants born to mothers who had been infected by measles wild virus (group I) with those born to mothers who had been vaccinated (group II). MATERIAL/METHODS: Serum samples were tested for measles antibodies from 79 infants in the 7th month of life and from 27 mothers between 17 and 41 years of age. Two commercial enzyme immunoassays (EIA) and a plaque neutralization test (PNT) were used. RESULTS: Only 12.7% of all infants showed measles antibodies in EIA. However, antibodies could be detected by PNT in all the infants, although only 36.6% showed titers of >1:8, which corresponds to protective antibody values of >0.2 IU/ml. The mean geometrical titer was significantly higher among infants from group I than in group II (1:7.16 vs. 1:3.71, p=0.0038). Protective antibody titers were detected in 50% of infants from group I and only 18.2% in group II (p<0.02). CONCLUSIONS: Passive acquired immunity in infants born to mothers who have had measles lasts longer than in infants born to vaccinated mothers. Nearly two thirds of infants (65.4%) in the 7th month of life did not have sufficient maternally derived neutralizing antibodies to protect against measles. Our data suggest that the recommended age for the first dose of measles vaccine during measles epidemics should be lowered to 9 months, with re-vaccination at 12-15 months.

Progress toward measles elimination in Germany.

While the former East Germany (FEG) achieved a reduction of measles incidence to <1 case per 100,000 population before reunification in 1990, the former West Germany (FWG) experienced significant measles morbidity. In 2001, according to statutory surveillance data, the incidence of measles was still higher in FWG than in FEG (8.7 vs. 0.7 cases/100,000 population). This article describes the development of the vaccination strategies in FEG and FWG, vaccination coverage, results of seroprevalence studies, measles surveillance in Germany, the epidemiology of a recent outbreak, and the role of laboratory diagnosis for measles control in Germany. Recent establishment of comprehensive nationwide surveillance and prevention programs to attain higher vaccine coverage have led to a decrease in measles incidence. However, further improvement of age-appropriate vaccine coverage and closure of immunity gaps in school-age children are necessary to eliminate measles in Germany.

Rapid replacement of endemic measles virus genotypes.

Although vaccination campaigns have significantly reduced the number of measles cases worldwide, endemic transmission of measles virus (MV) continues to occur in several continents, including Europe. To obtain current information on measles incidence and molecular data on circulating MVs in Germany, a nationwide measles sentinel was established. Phylogenetic analysis based on the variable part of the N gene from 80 MVs isolated between November 1999 and October 2001 revealed the presence of at least six distinct MV genotypes: B3, C2, D4, D6, G2 and a new variant of D7. Both the incidence and the pattern of MV genotypes differed markedly between the former East and West Germany. In the eastern part, few measles cases, mainly caused by genotypes originating from other countries (B3, D4, G2), were detected. In the western and southern parts, genotypes C2, D6 and D7 were associated with endemic transmission. Surprisingly, the indigenous genotypes predominant during the 1990s - C2 and D6 - disappeared simultaneously over the period of observation coinciding with the emergence and the wide spread of D7 viruses. While the incidence of measles remained constant, all MVs isolated in 2001 were assigned to D7. We note that the haemagglutinin (H) sequence of D7 viruses shows distinct exchanges of certain amino acids in the stem and propeller domain compared to C2, D6 and the MV vaccine strains used. This raises the possibility of a selective advantage of D7 viruses transmitted in the presence of H-specific antibodies.