Validation of monoplex assays detecting antibodies against Corynebacterium diphtheriae and Clostridium tetani toxins, rubella virus and parvovirus B19 for incorporation into Multiplex Serology.

Serological assays detecting antibodies in serum or plasma samples are useful and versatile instruments to investigate an individual's infection and vaccination history, e.g. for clinical diagnosis, personal risk evaluation, and seroepidemiological studies. Multiplex Serology is a suspension bead array-based high-throughput methodology for simultaneous measurement of antibodies against multiple pathogens in a single reaction vessel, thus economizing sample volume, measurement time, and costs. We developed and validated bead-based pathogen-specific Monoplex Serology assays, i.e. assays including only antigens for the respective pathogen, to detect antibodies against Corynebacterium diphtheriae and Clostridium tetani toxins, rubella virus and parvovirus B19. The developed assays expand the portfolio of existing pathogen-specific bead-based serology assays and can be efficiently incorporated into larger Multiplex Serology panels. The newly developed Monoplex Serology assays consist of only one antigen per infectious agent, expressed as Glutathione S-transferase-fusion proteins in E. coli. Specificity, sensitivity and Cohen's kappa statistics in comparison with routine clinical diagnostic assays were calculated for serum dilutions 1:100 and 1:1000. All pathogen-specific assays were successfully validated at both serum dilutions with the exception of rubella Monoplex Serology which showed impaired sensitivity (57.6%) at dilution 1:1000. Specificities of successfully validated Monoplex Serology assays ranged from 85.6% to 100.0% (median: 91.7%), and sensitivities from 81.3% to 95.8% (median: 90.9%); agreement with the reference assays ranged from substantial to almost perfect (kappa: 0.66-0.86, median: 0.78). Statistical performance and slim assay design enable efficient incorporation of the developed assays into Multiplex Serology.

Evaluation of two (02) platforms for chemiluminescence-based detection of anti-rubella IgG antibodies in a sub-Saharan country, Côte d'Ivoire.

The diagnosis of rubella is mainly made in pregnant women and the newborn by specific IgG and/or IgM detection. In addition to HAI and ELISA techniques, new immunoanalytical methods have been developed. This study aimed to evaluate two chemiluminescence platforms, Architect i2000SR and Maglumi 800 for rubella biological diagnosis in Côte d'Ivoire. Blood samples were taken from 113 pregnant women aged 15 to 30 in prenatal care. Samples were analyzed for Rubella IgG detection at the NBTS laboratory on the evaluated platforms and the Cobas e601 used as a reference. The majority of women were in their second trimester of pregnancy. Among them, only 13.3% were vaccinated against rubella. The evaluated platforms showed good precision with coefficients of variation >10%. Regarding analytical performances, sensitivities were 97.53% and 96.29% whereas specificities were 100% and 96.88% for Architect I2000SR and Maglumi800, respectively. Both platforms showed good agreement with cobas e601 for antibody levels <200 IU/ml and <350UI/ml for Architect and Maglumi 800, respectively. Findings of the current study revealed that the two platforms have similar features with Cobas e601 and could be used routinely for the serological diagnosis of rubella. However, the results of one platform should not be extrapolated to another.

Sample size calculation for estimating key epidemiological parameters using serological data and mathematical modelling.

BACKGROUND: Our work was motivated by the need to, given serum availability and/or financial resources, decide on which samples to test in a serum bank for different pathogens. Simulation-based sample size calculations were performed to determine the age-based sampling structures and optimal allocation of a given number of samples for testing across various age groups best suited to estimate key epidemiological parameters (e.g., seroprevalence or force of infection) with acceptable precision levels in a cross-sectional seroprevalence survey. METHODS: Statistical and mathematical models and three age-based sampling structures (survey-based structure, population-based structure, uniform structure) were used. Our calculations are based on Belgian serological survey data collected in 2001-2003 where testing was done, amongst others, for the presence of Immunoglobulin G antibodies against measles, mumps, and rubella, for which a national mass immunisation programme was introduced in 1985 in Belgium, and against varicella-zoster virus and parvovirus B19 for which the endemic equilibrium assumption is tenable in Belgium. RESULTS: The optimal age-based sampling structure to use in the sampling of a serological survey as well as the optimal allocation distribution varied depending on the epidemiological parameter of interest for a given infection and between infections. CONCLUSIONS: When estimating epidemiological parameters with acceptable levels of precision within the context of a single cross-sectional serological survey, attention should be given to the age-based sampling structure. Simulation-based sample size calculations in combination with mathematical modelling can be utilised for choosing the optimal allocation of a given number of samples over various age groups.

Measles and rubella seroprevalence among adults in Georgia in 2015: helping guide the elimination efforts.

A large-scale measles outbreak (11 495 reported cases, 60% aged ≥15 years) occurred in Georgia during 2013-2015. A nationwide, multistage, stratified cluster serosurvey for hepatitis B and C among persons aged ≥18 years conducted in Georgia in late 2015 provided an opportunity to assess measles and rubella (MR) susceptibility after the outbreak. Residual specimens from 3125 participants aged 18-50 years were tested for Immunoglobulin G antibodies against MR using ELISA. Nationwide, 6.3% (95% CI 4.9%-7.6%) of the surveyed population were seronegative for measles and 8.6% (95% CI 7.1%-10.1%) were seronegative for rubella. Measles susceptibility was highest among 18-24 year-olds (10.1%) and declined with age to 1.2% among 45-50 year-olds (P < 0.01). Susceptibility to rubella was highest among 25-29 year-olds (15.3%), followed by 18-24 year-olds (11.6%) and 30-34 year-olds (10.2%), and declined to <5% among persons aged ≥35 years (P < 0.001). The susceptibility profiles in the present serosurvey were consistent with the epidemiology of recent MR cases and the history of the immunization programme. Measles susceptibility levels >10% among 18-24 year-olds in Georgia revealed continued risk for outbreaks among young adults. High susceptibility to rubella among 18-34 year-olds indicates a continuing risk for congenital rubella cases.

Seroprevalence of measles, mumps & rubella antibodies among 5-10 years old children in north India.

Background & objectives: : Globally, there is an effort to eliminate the measles and control rubella as these diseases lead to considerable morbidity and mortality especially among under-five children and are important public health problems. This study was aimed to estimate the seroprevalence of measles, mumps and rubella (MMR) antibodies among children of age 5-10 yr in Chandigarh, north India, to provide evidence on prevalent immunity levels. Methods: : This cross-sectional study was conducted in Chandigarh, among 196 randomly selected healthy children (5-10 yr), who received either one or two doses of measles or MMR combination vaccine. Socio-economic background and immunization history were recorded. Blood sample (2 ml) was collected to estimate the MMR IgG antibody titres by using ELISA kits. Results: : Protective seroprevalence of MMR antibodies was 40.8, 75.5 and 86.2 per cent, respectively. The geometric mean titres of MMR IgG antibodies in the study children were 11.3, 50.6 and 54.3 international units (IU)/ ml, respectively. The proportion of seroprotected children for measles was significantly higher among those who had received two or more doses (46.4%) of measles vaccine compared to those who had received single dose (35.6%) (P <0.001). About 16 per cent of children had received single dose of MMR vaccine. Among these, 71.4 and 100 per cent were seroprotected against mumps and rubella, respectively. Interpretation & conclusions: : A large proportion of children aged 5-10 yr lacked protective immunity against measles (60%); about one-fourth (15-25%) were susceptible to infection with mumps and rubella virus. Mumps vaccination may be considered to be included in National Immunization Schedule for children with periodic serosurveillance.

Seasonal influence on TORCH infection and analysis of multi-positive samples with indirect immunofluorescence assay.

BACKGROUND: TORCH including the pathogens of Toxoplasma gondii (TOX), rubella virus (RV), cytomegalovirus (CMV), and herpes simplex virus (HSV) causes intrauterine infections and poses a worldwide threat to women especially in pregnancy. In this study, we described the seasonal difference in TORCH infection and analyzed the anti-TORCH IgM multipositive serum samples by the indirect immunofluorescence assays (IFA). METHODS: To observe the seasonal influence of the anti-TORCH IgG and IgM antibodies, a retrospective study was conducted with 10 669 women (20-40 y old) before pregnancy from August 2016 to July 2017. Totally 199 ELISA anti-TORCH IgM multipositive serum samples were further tested by IFAs for false-positive analysis. RESULTS: The prevalence of positive HSV1-IgM, RV-IgM, HSV2-IgM, CMV-IgM, and TOX-IgM in the present population was 6.30%, 2.55%, 1.94%, 1.24%, and 0.67%, respectively. Additionally, the prevalence of positive RV-IgM, CMV-IgM, and HSV1-IgM was statistically different among four seasons, with the highest positive rates of RV-IgM (4.12%) in autumn, CMV-IgM (1.75%) in summer, and HSV1-IgM (7.53%) in winter. The confirmatory IFAs showed that the positive rates of RUV-IgM, CMV-IgM, and HSV2-IgM were significantly different from those in ELISA screening experiments. Interestingly, only 32.7% (65/199) of the TORCH IgM multipositive results were consistent with those by the IFA, indicating that cross-reaction caused false positives were common in ELISA IgM antibody screening. CONCLUSION: The TORCH infection displayed different prevalence among four seasons in our 12-month retrospective study. The IgM multipositives by ELISA screening may need further confirmation analysis due to its relatively high cross-reaction rate.

Comparison of the LIAISON®XL and ARCHITECT IgG, IgM, and IgG avidity assays for the diagnosis of Toxoplasma, cytomegalovirus, and rubella virus infections.

This study compared the performance of the LIAISON®XL system of immunoglobulin (Ig) G and IgM immunoassays for the diagnosis of Toxoplasma gondii, cytomegalovirus (CMV), and rubella virus infections with that of the ARCHITECT system. Patient serum samples, previously screened and clinically diagnosed with T. gondii, CMV or rubella, were used to compare LIAISON®XL and ARCHITECT IgG and IgM immunoassays. LIAISON®XL Toxo and CMV IgG avidity assays were also compared with equivalent ARCHITECT assays and reference methods. Overall agreement between the LIAISON®XL and ARCHITECT assays was 99% and 92% for the Toxo IgG and IgM assays, respectively, 98% and 96% for the CMV IgG and IgM assays, respectively, and 93% and 98% for the rubella virus IgG and IgM assays, respectively. LIAISON®XL IgG Toxo and CMV avidity assays showed high concordance with the VIDAS® Toxo IgG avidity assay and an in-house CMV avidity assay (reference methods), and faster IgG avidity maturation in a larger number of samples collected months after the primary infection compared with equivalent ARCHITECT assays. LIAISON®XL assays for detection of anti-T. gondii, CMV and rubella virus IgG and IgM are at least equal to the competitor assays on the ARCHITECT platform.

Diagnostic algorithm for the confirmation of cases of measles and rubella in Mexico.

Due to the successful implementation of measles and rubella elimination strategies, Mexico announced the interruption of endemic transmission of measles in 1996 and that of rubella in 2008. After a verification process, the region of the Americas was declared free of rubella and congenital rubella syndrome in 2015 and of measles in 2016. In order to maintain the elimination status in Mexico, it is essential to continue laboratory surveillance within the framework of the Global Measles and Rubella Laboratory Network. The Institute of Epidemiological Diagnosis and Reference, through the National Network of Public Health Laboratories, guarantees timely and reliable results in view of the possible reintroduction of these and other emerging pathogens.

Association between false-positive TORCH and antiphospholipid antibodies in healthy pregnant women.

The present study investigated: (a) the presence of antiphospholipid antibodies and (b) the obstetric outcome in healthy pregnant women showing false-positive TORCH-Toxoplasmosis, Other: syphilis, varicella-zoster, Rubella, Cytomegalovirus (CMV), and Herpes infections-results. Data from 23 singleton healthy pregnancies with false-positive TORCH results were collected. Each woman was systematically screened for TORCH IgG and IgM during the pre-conception assessment and/or at the beginning of pregnancy. In the presence of IgM positivity, when indicated (CMV, toxoplasmosis, rubella, herpes simplex virus), IgG avidity was evaluated and, if possible, polymerase chain reaction was performed on an amniotic fluid sample in order to distinguish between primary infection or false positivity. The antiphospholipid antibodies tests were: lupus anticoagulant, anticardiolipin antibodies IgG, IgM, and anti-ß2glicoprotein I IgG, IgM. The antiphospholipid antibodies tests, if positive, were repeated after 12 weeks to confirm the results. In pregnant women with false-positive TORCH, the overall prevalence of positive antiphospholipid antibodies for one or more tests was 52.2%. To clarify the correlation of false-positive TORCH results with clinical practice, obstetric outcome was analyzed in terms of live births, week of delivery, neonatal birth weight, and neonatal birth weight percentile. A statistically significant lower neonatal birth weight and neonatal birth weight percentile were observed in women with false-positive TORCH associated with antiphospholipid antibodies positivity (Group A) in comparison with those in women with false-positive TORCH without antiphospholipid antibodies positivity (Group B). No statistically significant difference was found for the week of delivery between the two groups. It is hoped that future studies will verify the life-long persistence of antiphospholipid antibodies positivity by follow-up of these women and identify who will develop a classical antiphospholipid syndrome or other autoimmune disorders.

Seroprevalence of HIV, HTLV, CMV, HBV and rubella virus infections in pregnant adolescents who received care in the city of Belém, Pará, Northern Brazil.

BACKGROUND: Prenatal tests are important for prevention of vertical transmission of various infectious agents. The objective of this study was to describe the prevalence of human immunodeficiency virus (HIV), human T-lymphotropic virus (HTLV), hepatitis B virus (HBV), cytomegalovirus (CMV), rubella virus and vaccination coverage against HBV in pregnant adolescents who received care in the city of Belém, Pará, Brazil. METHODS: A cross-sectional study was performed with 324 pregnant adolescents from 2009 to 2010. After the interview and blood collection, the patients were screened for antibodies and/or antigens against HIV-1/2, HTLV-1/2, CMV, rubella virus and HBV. The epidemiological variables were demonstrated using descriptive statistics with the G, χ2 and Fisher exact tests. RESULTS: The mean age of the participants was 15.8 years, and the majority (65.4%) had less than 6 years of education. The mean age at first intercourse was 14.4 years, and 60.8% reported having a partner aged between 12 and 14 years. The prevalence of HIV infection was 0.3%, and of HTLV infection was 0.6%. Regarding HBV, 0.6% of the participants had acute infection, 9.9% had a previous infection, 16.7% had vaccine immunity and 72.8% were susceptible to infection. The presence of anti-HBs was greater in adolescent between 12 and 14 years old (28.8%) while the anti-HBc was greater in adolescent between 15 and 18 years old (10.3%). Most of the adolescents presented the IgG antibody to CMV (96.3%) and rubella (92.3%). None of the participants had acute rubella infection, and 2.2% had anti-CMV IgM. CONCLUSIONS: This study is the first report of the seroepidemiology of infectious agents in a population of pregnant adolescents in the Northern region of Brazil. Most of the adolescents had low levels of education, were susceptible to HBV infection and had IgG antibodies to CMV and rubella virus. The prevalence of HBV, HIV and HTLV was similar to that reported in other regions of Brazil. However, the presence of these agents in this younger population reinforces the need for good prenatal follow-up and more comprehensive vaccination campaigns against HBV due to the large number of women susceptible to the virus.

Vaccination Status and Antibody Titers against Rubella and Measles among Japanese Female College Students Majoring in Childcare between 2015 and 2018.

In 2014, for the protection of medical workers against measles and rubella infection, the Japanese Society for Infection Prevention and Control (JSIPC) recommended either maintaining antibody titers of seroprotective range or two-dose vaccination. JSIPC defined antibody titers into 3 ranges: seroprotective as expected prevention of infection, seronegative as under detection levels, and seropositive as antibody titers ranged between seronegative and seroprotective. This study aimed to explore the association between the number of vaccine doses received and the antibody titers against measles and rubella among Japanese college students majoring in childcare. A total of 841 female students with no history of measles or rubella were serologically screened at the time of college admission between 2015 and 2018. All 841 students had been vaccinated against measles; 738 (87.8%) received two doses of the measles vaccine and 103 (12.2%) received one dose. Likewise, 839 students, except for two, had been vaccinated against rubella; 719 (85.7%) received two doses of the rubella vaccine and 120 (14.3%) received one dose. We thus found that 107 students (12.7%) were seropositive for measles-specific IgG and 731 (86.9%) attained seroprotective titers. By contrast, in case of rubella-specific IgG, only 462 students (55.1%) attained seroprotective titers, and 371 students (44.1%) were seropositive. The two students without receiving rubella vaccination were classified as seronegative. In conclusion, despite that > 85% of students surveyed had received two doses of measles and rubella vaccines, a substantial number of students remain susceptible to measles and especially rubella at the time of college admission.

Lack of immunity against rubella among Italian young adults.

BACKGROUND: To support the evaluation of the 2010-15 National Plan for Measles and Congenital Rubella Elimination, the authors designed and performed a serosurveillance survey to verify the immunity/susceptibility rate against rubella among Apulian young adults. METHODS: The study was carried out from May 2011 to June 2012 in the Department of Transfusion Medicine/Blood Bank of Policlinico General Hospital in Bari. Subjects were enrolled by a convenience sampling. For each enrolled patient a 5 ml serum sample was collected and tested for anti-rubella IgG. The geometrical means (GMT) of anti-rubella IgG was calculated. T student test or ANOVA test, when appropriate, was used to compare the means of age per gender and GMT of anti-rubella IgG titres per age classes. Chi-square test was used to compare the proportion of anti-rubella IgG positive subjects per gender and per age classes. For all tests, a p value <0.05 was considered as significant. RESULTS: At the end of the study 1764 subjects were enrolled, 1362 (77.2%) of which were male. The mean age was 38.4 ± 11.7 years (range: 17-65). 86.7% (95% CI = 85.0-88.2) had a positive titre of anti-rubella IgG. GMT of anti-rubella IgG titre was 4.3. The proportion of positive subjects was of 76.8% (n = 279/363; 95% CI = 72.2-81.1) in persons aged 18-26 years; 88.1% (n = 310/352; 95% CI = 84.2-91.3) in 27-35 year-old people; 88.5% (n = 464/524; 95% CI = 85.5-91.1) in 36-45 year-old people; 90.7% (n = 350/386; 95% CI = 87.3-93.4) among people aged 46-55 years and 90.6% (n = 126/139; 95% CI = 84.5-94.9) in 55-65 year-old people (Chi-square = 39.7; p < 0.0001). GMT of anti-rubella IgG titre was 4.3 (4.3 in male and 4.2 in female, t = 2.2; p = 0.03) and seems to differ dividing the enrolled subjects by age group (F = 14.3; p < 0.0001). CONCLUSIONS: According to our data, too many women of child-bearing age are still unprotected from rubella in the elimination era and in this scenario the public health efforts should be oriented to catch-up activities.

Rubella in Sub-Saharan Africa and sensorineural hearing loss: a case control study.

BACKGROUND: Rubella infection can affect several organs and cause birth defects that are responsible for congenital rubella syndrome (CRS). Congenital hearing loss is the most common symptom of this syndrome, occurring in approximately 60% of CRS cases. Worldwide, over 100 000 babies are born with CRS every year. There is no specific treatment for rubella, but the disease is preventable by vaccination. Since 1969, the rubella vaccine has been implemented in many countries, but in Africa, only a few countries routinely immunize against rubella. The aim of this study was to estimate the rate of infection from the wild-type rubella virus in São Tomé and Príncipe by determining rubella seroprevalence with a DBS method. The goal of this study was to reinforce the need for implementation of the rubella vaccine in this country. As secondary objectives, the validation of a DBS method was first attempted and an association between seroprevalence and hearing loss was assessed. METHODS: We collected samples from individuals observed during humanitarian missions in São Tomé and Príncipe. All individuals underwent an audiometric evaluation, and a drop of blood was collected for the dried blood spot (DBS). We define two groups: the case group (individuals with unilateral or bilateral hearing loss (HL)) and the control group (individuals with two normal ears). Patients were excluded if they suffered from conductive HL, if they showed evidence of possible causes of HL, if they had developmental delay or if they refused to participate in the study. RESULTS: Among the 315 subjects, we found 64.1% individuals with IgG for the rubella virus, 32.1% without immunity for the rubella virus and 3.8% who were borderline. In the control group, 62.6% were positive for the rubella IgG, whereas in the case group, 72% were positive. Analyzing both groups, with ages ranging from 2 to 14 years of age and from 15 to 35 years of age, we found a seroprevalence of 50.3% to rubella in the younger group and 82.1% in the older group, with a significant difference between cases and control group noted within the younger patients (p = 0.025). CONCLUSIONS: Rubella is a disease that can be prevented. Rubella infections are still very common in São Tomé and Príncipe, and women of child-bearing age are still at risk for rubella infection during pregnancy, justifying the urgency of vaccination against rubella. A statistically significant association between the group of children under 14 years of age with HL and immunity for rubella was observed in this country, although this study did not allow us to establish a cause-effect relationship between rubella infection and SNHL.

Genotypes of rubella virus and the epidemiology of rubella infections in the Democratic Republic of the Congo, 2004-2013.

Rubella is a viral infection that may cause fetal death or congenital defects, known as congenital rubella syndrome (CRS), during early pregnancy. The World Health Organization (WHO) recommends that countries assess the burden of rubella and CRS, including the determination of genotypes of circulating viruses. The goal of this study was to identify the genotypes of rubella viruses in the Democratic Republic of the Congo (DRC). Serum or throat swab samples were collected through the measles surveillance system. Sera that tested negative for measles IgM antibody were tested for rubella IgM antibody. Serum collected within 4 days of rash onset and throat swabs were screened by real-time RT-PCR for rubella virus RNA. For positive samples, an amplicon of the E1 glycoprotein gene was amplified by RT-PCR and sequenced. 11733 sera were tested for rubella IgM and 2816 (24%) were positive; 145 (5%) were tested for the presence of rubella RNA by real-time RT-PCR and 10 (7%) were positive. Seventeen throat swabs were analyzed by RT-PCR and three were positive. Sequences were obtained from eight of the positive samples. Phylogenetic analysis showed that the DRC rubella viruses belonged to genotypes 1B, 1E, 1G, and 2B. This report provides the first information on the genotypes of rubella virus circulating in the DRC. These data contribute to a better understanding of rubella burden and the dynamics of rubella virus circulation in Africa. Efforts to establish rubella surveillance in the DRC are needed to support rubella elimination in Africa. J. Med. Virol. 88:1677-1684, 2016. © 2016 Wiley Periodicals, Inc.

Measles & rubella outbreaks in Maharashtra State, India.

BACKGROUND & OBJECTIVES: Under the outbreak-based measles surveillance in Maharashtra State the National Institute of Virology at Pune receives 3-5 serum samples from each outbreak and samples from the local hospitals in Pune for laboratory diagnosis. This report describes one year data on the measles and rubella serology, virus isolation and genotyping. METHODS: Maharashtra State Health Agencies investigated 98 suspected outbreaks between January-December 2013 in the 20 districts. Altogether, 491 serum samples were received from 20 districts and 126 suspected cases from local hospitals. Samples were tested for the measles and rubella IgM antibodies by commercial enzyme immunoassay (EIA). To understand the diagnostic utility, a subset of serum samples (n=53) was tested by measles focus reduction neutralization test (FRNT). Further, 37 throat swabs and 32 urine specimens were tested by measles reverse transcription (RT)-PCR and positive products were sequenced. Virus isolation was performed in Vero hSLAM cells. RESULTS: Of the 98 suspected measles outbreaks, 61 were confirmed as measles, 12 as rubella and 21 confirmed as the mixed outbreaks. Four outbreaks remained unconfirmed. Of the 126 cases from the local hospitals, 91 were confirmed for measles and three for rubella. Overall, 93.6 per cent (383/409) confirmed measles cases were in the age group of 0-15 yr. Measles virus was detected in 18 of 38 specimens obtained from the suspected cases. Sequencing of PCR products revealed circulation of D4 (n=9) and D8 (n=9) strains. Four measles viruses (three D4 & one D8) were isolated. INTERPRETATION & CONCLUSIONS: Altogether, 94 measles and rubella outbreaks were confirmed in 2013 in the State of Maharasthra indicating the necessity to increase measles vaccine coverage in the State.

Some factors affecting rubella seronegative prevalence among pregnant women in a North West England region between April 2011 and March 2013.

BACKGROUND: Rubella is usually a mild viral illness, but during pregnancy, it can have potentially devastating effects causing fetal losses and severe congenital malformations (congenital rubella syndrome). Rubella is now rare in most developed countries following a successful vaccination programme. We aimed to investigate differences in epidemiological profile of pregnant women screened antenatally in Liverpool to identify risk factors for rubella immunity. METHODS: All samples were tested with the Elecsys Rubella IgG immunoassay kit. A result <10 IU/ml was considered to be seronegative. RESULTS: The seronegativity prevalence among pregnant women in Liverpool (6.3%) is higher than average value for the North West region (3.7%). The seronegative rates varied with age (15.4% for <15 years, 18.7% for 15-20 years, compared with 2% for 30-35 years). The areas with the highest seronegative rates correspond with areas of Liverpool with high pockets of socioeconomic deprivation. CONCLUSION: The highest proportion of seronegative women were among the youngest age groups. Local areas with highest level of deprivation should be given priority and additional resources to develop targeted programmes and pathways to implement appropriate interventions such as MMR catch-up programmes and put in place arrangements for offering MMR vaccination in maternity units.

Epidemiology of rubella virus cases in the pre-vaccination era of Ethiopia, 2009-2015.

BACKGROUND: Rubella is a common mild rash illness caused by rubella virus. The majority of infections occur in children and young adults. The infection is the cause of a serious birth defect known as Congenital Rubella Syndrome (CRS) when a woman acquires infection early in pregnancy. Ethiopia has not yet established rubella virus surveillance and has not yet introduced rubella vaccine into the routine immunization program. We characterize the epidemiology of laboratory confirmed rubella virus cases collected through measles surveillance from 2009 to 2015 to better understand the burden of the disease in the country. METHODS: A descriptive analysis was made to characterize rubella cases reported through the national measles case based surveillance system. The measles case definition was used to capture potential rubella cases. A suspected measles case was a person with generalized rash and fever with cough, or coryza or conjunctivitis. Those cases whose sera were negative for measles IgM antibodies were tested for rubella IgM antibody. A confirmed rubella case was a person who tested positive for rubella IgM. Only laboratory confirmed rubella cases were analyzed in this article. RESULTS: Between 2009 and 2015, a total of 28,284 serum/plasma samples were collected and tested for measles IgM antibody and 11,151 (39.4%) were found positive. A total of 17,066 measles IgM negative or indeterminate samples were tested for rubella virus IgM and 2615 (15.3%) were found positive during the same period. Of 2615 confirmed rubella cases, 52.2% were females. The age of confirmed cases ranged from one month to 42 years with a mean age of 7.3 years. Three-fourth of all confirmed rubella cases were aged less than 10 years. The number of laboratory confirmed rubella cases linearly increased from 83 in 2009 to 856 in 2013 but dropped to 222 and 319 in 2014 and 2015 respectively. Higher number of cases occurred in the hot dry season (January through June) and in the central and western part of Ethiopia with 127 lab-confirmed outbreaks in the study period. CONCLUSIONS: Based on our analysis, rubella was found to be endemic throughout Ethiopia. Children below the age of 10 years were the most affected. The burden of rubella cases varied from year to year but had a seasonal peak in March. To better understand the magnitude of rubella prior to vaccine introduction, establishing rubella surveillance system, conducting sero-prevalence studies among child bearing age females and establishing CRS sentinel surveillance among young infants are critical.

Digital microfluidic platform for the detection of rubella infection and immunity: a proof of concept.

BACKGROUND: Whereas disease surveillance for infectious diseases such as rubella is important, it is critical to identify pregnant women at risk of passing rubella to their offspring, which can be fatal and can result in congenital rubella syndrome (CRS). The traditional centralized model for diagnosing rubella is cost-prohibitive in resource-limited settings, representing a major obstacle to the prevention of CRS. As a step toward decentralized diagnostic systems, we developed a proof-of-concept digital microfluidic (DMF) diagnostic platform that possesses the flexibility and performance of automated immunoassay platforms used in central facilities, but with a form factor the size of a shoebox. METHODS: DMF immunoassays were developed with integrated sample preparation for the detection of rubella virus (RV) IgG and IgM. The performance (sensitivity and specificity) of the assays was evaluated with serum and plasma samples from a commercial antirubella mixed-titer performance panel. RESULTS: The new platform performed the essential processing steps, including sample aliquoting for 4 parallel assays, sample dilution, and IgG blocking. Testing of performance panel samples yielded diagnostic sensitivity and specificity of 100% and 100% for both RV IgG and RV IgM. With 1.8 µL sample per assay, 4 parallel assays were performed in approximately 30 min with <10% mean CV. CONCLUSIONS: This proof of concept establishes DMF-powered immunoassays as being potentially useful for the diagnosis of infectious disease.

Epidemiological Evaluation of Rubella Virus Infection among Pregnant Women in Ibadan, Nigeria.

Rubella is a vaccine-preventable, mild rash-inducing viral disease with complications that include a spectrum of birth defects in the developing fetus, especially if the infection is acquired in the early months of pregnancy. Consequently, the primary objective of global rubella control programs is prevention of congenital rubella infection and associated birth defects. Despite the availability of safe and effective vaccines, and the elimination of the rubella virus in many developed countries, substantial commitment to rubella control has not been demonstrated in developing countries. This study appraises immunity to rubella, and consequently makes appropriate recommendations aimed at facilitating effective control. A cross-sectional sero-surveillance study was carried out among defined 272 consenting ante-natal clinic attendees in south-western, Nigeria. Prevalence rates of 91.54% and 1.84% were recorded for the anti-rubella virus (anti-RV) IgG and IgM, respectively. Also, 90.7% and 92.3% of the women aged ≤30 years and >30 years, respectively, had detectable anti-RV IgG. No significant association (p = 0.94) was recorded between anti-RV IgG detection and age of the women. Previous exposure and susceptibility of significant fraction of the population to rubella infection were confirmed. Considerable political commitment and promotion of free rubella immunization specifically for women with childbearing potential were recommended.

[The multiplex method of estimation of humoral immunity to vaccine regulated childhood infections].

The goal of this work was to present the results of the laboratory tests of the multiplex dot immunoassay method using protein microarray for complex estimation of humoral immunity to measles, mumps, and rubella viruses. It was shown that the obtained results were in a good agreement with data of commercial monospecific ELISA kits. The developed method is fast, requires fewer resources, and may be used in the field.