[show abstract][hide abstract] ABSTRACT: Following human infections with novel avian influenza
A(H7N9) viruses in China, the European Centre
for Disease Prevention and Control, the World Health
Organization (WHO) Regional Office for Europe and
the European Reference Laboratory Network for
Human Influenza (ERLI-Net) rapidly posted relevant
information, including real-time RT-PCR protocols.
An influenza RNA sequence-based computational
assessment of detection capabilities for this virus was
conducted in 32 national influenza reference laboratories
in 29 countries, mostly WHO National Influenza
Centres participating in the WHO Global Influenza
Surveillance and Response System (GISRS). Twentyseven
countries considered their generic influenza A
virus detection assay to be appropriate for the novel
A(H7N9) viruses. Twenty-two countries reported having
containment facilities suitable for its isolation and
propagation. Laboratories in 27 countries had applied
specific H7 real-time RT-PCR assays and 20 countries
had N9 assays in place. Positive control virus RNA was
provided by the WHO Collaborating Centre in London
to 34 laboratories in 22 countries to allow evaluation
of their assays. Performance of the generic influenza A
virus detection and H7 and N9 subtyping assays was
good in 24 laboratories in 19 countries. The survey
showed that ERLI-Net laboratories had rapidly developed
and verified good capability to detect the novel
A(H7N9) influenza viruses.
Euro surveillance: bulletin europeen sur les maladies transmissibles = European communicable disease bulletin 01/2014; 19(4). · 5.49 Impact Factor
[show abstract][hide abstract] ABSTRACT: Background. Middle East Respiratory Syndrome Coronavirus (MERS-CoV) emerged in 2012 causing Severe Acute Respiratory Disease and pneumonia, with ∼44% mortality in 136 cases till date. Design of vaccines to limit the virus spread or diagnostics to track newly emerging strains requires knowledge of antigenic and serological relationships of MERS-CoV to other coronaviruses.Methods. Using synthetic genomics and Venezuelan Equine Encephalitis Virus replicons (VRPs) expressing S and N proteins from MERS-CoV and other human and bat CoV`s, we characterize the antigenic (using western blots and ELISA) and serological responses (using Neutralization Assays) against two MERS-CoV isolates in comparison with other human and bat coronaviruses.Results. Serologic and neutralization responses against the S glycoprotein were primarily strain specific with very low level cross reactivity within, or across subgroups. Coronaviruses N proteins within, but not across subgroups, share cross-reactive epitopes with MERS-CoV isolates. Our findings were validated using MERS-CoV patient (NA 01) convalescent serum, and human serum to SARS-CoV, NL63 and OC43.Conclusions. Vaccine design for emerging coronaviruses should involve chimeric S protein containing neutralizing epitopes from multiple virus strains across subgroups, to reduce immune pathology, and diagnostic platform should include a panel of N and S proteins from phylogenetically distinct coronaviruses.
The Journal of Infectious Diseases 11/2013; · 5.85 Impact Factor
[show abstract][hide abstract] ABSTRACT: Background. We report a follow up clinical and serological investigation of 274 children receiving seasonal influenza vaccine (TIV) one year after receipt of either AS03B-adjuvanted subunit or whole virus monovalent A(H1N1)pdm09 vaccine and describe the antibody responses to the H3N2 A/Perth/16/2009 and B/Brisbane/60/2008 components of TIV. Method and Findings. Vaccine responses were analysed using Haemagglutination Inhibition (HAI) assays. In individuals under 3 years of age, previous receipt of adjuvanted vaccine resulted in higher HAI antibody responses to H3N2 and B strains compared to non-adjuvanted vaccine (fold change 16.8 vs 4.3 for H3N2 and 7.0 vs 1.6 for B). In older children, responses to the H3 and B components of TIV were similar between vaccine groups. Sera taken pre- and post pandemic vaccine were also analysed by HAI with A/Perth/16/2009 virus. This analysis showed that 11.1% of children receiving the AS03B-adjuvanted vaccine but only 1.4% in the non-adjuvanted group had a 4-fold rise to A/Perth/16/2009. Conculsion. AS03B-adjuvanted A(H1N1)pdm09 influenza vaccine generates a cross-reactive antibody response to H3N2 in children and enhances responses to heterologous subtypes in <3-year-old children one year later.
[show abstract][hide abstract] ABSTRACT: Current seasonal influenza vaccines have reduced immunogenicity and are of suboptimal efficacy in older adults. We have previously shown that the novel candidate vaccineMVA-NP+M1 is able to boost memory T cell responses in adultsaged 50-85 years. Pre-clinical studies have demonstrated that viral vectored vaccines can act as adjuvants when co-administered with protein-based vaccines.We have conducted a phase I clinical trial to compare co-administration of seasonal influenza vaccine and MVA-NP+M1, to seasonal influenza vaccine alone in adults aged 50 years and over. This combination of vaccines was safe and well tolerated. T cell responses to internal influenza proteins were boosted to significantly higher levels in the group receiving MVA-NP+M1 compared to the group receiving seasonal influenza vaccine alone. Rates of seroprotection and seroconversionagainst the three vaccine strains were similar in both groups however there was a significant increase in the geometric mean titre ratio for the H3N2 component of seasonal influenza vaccine in the co-administration group. While some vaccine combinations result in immune interference, the co-administration of MVA-NP+M1 alongside seasonal influenza vaccine is shown here to increase some influenza strain-specific antibody responses and boost memory T cells capable of recognising a range of influenza A subtypes.Molecular Therapy (2013); doi:10.1038/mt.2013.162.
[show abstract][hide abstract] ABSTRACT: Booster vaccination against 2009 H1N1 influenza virus was recommended for rheumatologic patients under immunosuppressive therapy during the 2009/2010 H1N1 pandemic. In this study we assessed whether B cell depletion with Rituximab influences of the antiviral immune response in 2009 H1N1 influenza virus-vaccinated patients.
Influenza virus-specific immune responses were analysed after the first and a booster vaccination with PandemrixTM in sixteen consecutive Rituximab-treated patients with different rheumatic autoimmune disorders. Antibody titers were determined by a haemagglutination-inhibition assay and virus-specific T cell responses were evaluated by a flow cytometry-based intracellular cytokine-secretion assay. Patients showing clinical symptoms of influenza infection were excluded from this study.
Two out of seven patients with low (<10%) and four out of nine with normal (>10%) B cells developed significant antibody responses after the first vaccination. Booster vaccination led to an antibody response in one additional patient. After the first vaccination, virus-specific CD4+ and CD8+ T cell responses were significantly lower in patients with low B cells than in those with normal B cells. Of importance, the booster vaccination stimulated the antiviral T cell response only in patients with low B cells.
In the absence of a significant effect of booster vaccinations against 2009 H1N1 influenza virus on the humoral immune response in B cell-depleted patients with autoimmune rheumatic diseases, enhanced antiviral T cell responses in patients with low B cells indicate that T cells, maybe, compensate for the impaired humoral immunity in these patients.
Clinical and experimental rheumatology 06/2013; · 2.66 Impact Factor
[show abstract][hide abstract] ABSTRACT: OBJECTIVE: To perform antiviral susceptibility monitoring of treated individuals in the community during the 2009 influenza A(H1N1) pandemic in England. PATIENTS AND METHODS: Between 200 and 400 patients were enrolled daily through the National Pandemic Flu Service (NPFS) and issued with a self-sampling kit. Initially, only persons aged 16 and over were eligible, but from 12 November (week 45), self-sampling was extended to include school-age children (5 years and older). All samples received were screened for influenza A(H1N1)pdm09 as well as seasonal influenza [A(H1N1), A(H3N2) and influenza B] by a combination of RT-PCR and virus isolation methods. Influenza A(H1N1)pdm09 RT-PCR-positive samples were screened for the oseltamivir resistance-inducing H275Y substitution, and a subset of samples also underwent phenotypic antiviral susceptibility testing by enzyme inhibition assay. RESULTS: We were able to detect virus by RT-PCR in self-taken samples and recovered infectious virus enabling further virological characterization. The majority of influenza A(H1N1)pdm09 RT-PCR-positive NPFS samples (n = 1273) were taken after oseltamivir treatment had begun. No reduction in phenotypic susceptibility to neuraminidase inhibitors was detected, but five cases with minority quasi-species of oseltamivir-resistant virus (an H275Y amino acid substitution in neuraminidase) were detected. CONCLUSIONS: Self-sampling is a useful tool for community surveillance, particularly for the follow-up of drug-treated patients. The virological study of self-taken samples from the NPFS provided a unique opportunity to evaluate the emergence of oseltamivir resistance in treated individuals with mild illness in the community, a target population that may not be captured by traditional sentinel surveillance schemes.
Journal of Antimicrobial Chemotherapy 06/2013; · 5.34 Impact Factor
[show abstract][hide abstract] ABSTRACT: During the summer of 2012, in Jeddah, Saudi Arabia a hitherto unknown coronavirus was isolated from the sputum of a patient with acute pneumonia and renal failure (1, 2).…
[show abstract][hide abstract] ABSTRACT: INTRODUCTION: Illness and death from influenza increase during pregnancy. In the United Kingdom pregnant women were targeted in a national programme for vaccination during the H1N1 2009-10 pandemic. METHODS: In this study, pregnant women were recruited in labour from November 9, 2009 to March 10, 2010. Pandemic vaccination status was determined. Venous cord blood collected at delivery was evaluated for transplacental transfer of antibodies by measurement of haemagglutination inhibition and microneutralization titres. RESULTS: Samples were collected from 77 vaccinated and 27 unvaccinated women. Seroprotection (HI titre ≥1∶40) was detected in 58 (75.3%, 95% CI 64.2-84.4) cord blood samples from vaccinated women and 5 (18.5%, 95% CI 6.3-38.1) from unvaccinated women (P<0.0001). There was evidence of transplacental seroprotection 8 days after maternal immunization (77.9%, 95 CI 66.2-87.1), maintained in most cases for at least 16 weeks. DISCUSSION: Immunization of pregnant women with AS03(A)-adjuvanted vaccine is followed by transplacental transfer of passive immunity at titres consistent with clinical protection in three-quarters of new-born infants. The findings support national and international pandemic H1N1 2009 recommendations for immunization during pregnancy.
PLoS ONE 01/2013; 8(1):e47448. · 3.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: Following the discovery in September 2012 of 2 patients, both with links to the Eastern Mediterranean Region, with serious respiratory illness due to novel coronavirus, all countries have instigated surveillance and laboratory activities to detect further cases, with intensive case-contact investigations undertaken on laboratory confirmation of cases. A total of 30 cases, of whom 18 have died, and at least 3 clusters have been detected to date (1 cluster among health-care workers and another 2 clusters among family members). To date, transmission studies have shown a low risk of onward human transmission, with clinical presentation remaining severe for the majority. Many questions remain including the zoonotic source and geographical extent of infection. Surveillance has been extended to include clusters of cases or health-care workers with severe, undiagnosed respiratory illness regardless of travel history. Environmental studies, on-going surveillance and linked case-contact investigations will provide a critical role in answering some of these issues.
Eastern Mediterranean health journal = La revue de santé de la Méditerranée orientale = al-Majallah al-ṣiḥḥīyah li-sharq al-mutawassiṭ 01/2013; 19 Suppl 1:S55-60.
[show abstract][hide abstract] ABSTRACT: Full text available at: http://currents.plos.org/outbreaks/article/state-of-knowledge-and-data-gaps-of-middle-east-respiratory-syndrome-coronavirus-mers-cov-in-humans-2/
Between September 2012 and 22 October 2013, 144 laboratory-confirmed and 17 probable MERS-CoV cases from nine countries were notified to WHO.
We summarize what is known about the epidemiology, virology, phylogeny and emergence of MERS-CoV to inform public health policies.
The median age of patients (n=161) was 50 years (range 14 months to 94 years), 64.5% were male and 63.4% experienced severe respiratory disease. 76.0% of patients were reported to have ≥1 underlying medical condition and fatal cases, compared to recovered or asymptomatic cases were more likely to have an underlying condition (86.8% vs. 42.4%, p<0.001). Analysis of genetic sequence data suggests multiple independent introductions into human populations and modelled estimates using epidemiologic and genetic data suggest R<sub>0</sub> is <1, though the upper range of estimates may exceed 1. Index/sporadic cases (cases with no epidemiologic-link to other cases) were more likely to be older (median 59.0 years vs. 43.0 years, p<0.001) compared to secondary cases, although these proportions have declined over time. 80.9% vs. 67.2% of index/sporadic and secondary cases, respectively, reported ≥1 underlying condition. Clinical presentation ranges from asymptomatic to severe pneumonia with acute respiratory distress syndrome and multi-organ failure. Nearly all symptomatic patients presented with respiratory symptoms and 1/3 of patients also had gastrointestinal symptoms.
Sustained human-to-human transmission of MERS-CoV has not been observed. Outbreaks have been extinguished without overly aggressive isolation and quarantine suggesting that transmission of virus may be stopped with implementation of appropriate infection control measures.
[show abstract][hide abstract] ABSTRACT: Very different influenza seasons have been observed from 2008/09-2011/12 in England and Wales, with the reported burden varying overall and by age group. The objective of this study was to estimate the impact of influenza on all-cause and cause-specific mortality during this period. Age-specific generalised linear regression models fitted with an identity link were developed, modelling weekly influenza activity through multiplying clinical influenza-like illness consultation rates with proportion of samples positive for influenza A or B. To adjust for confounding factors, a similar activity indicator was calculated for Respiratory Syncytial Virus. Extreme temperature and seasonal trend were controlled for. Following a severe influenza season in 2008/09 in 65+yr olds (estimated excess of 13,058 influenza A all-cause deaths), attributed all-cause mortality was not significant during the 2009 pandemic in this age group and comparatively low levels of influenza A mortality were seen in post-pandemic seasons. The age shift of the burden of seasonal influenza from the elderly to young adults during the pandemic continued into 2010/11; a comparatively larger impact was seen with the same circulating A(H1N1)pdm09 strain, with the burden of influenza A all-cause excess mortality in 15-64 yr olds the largest reported during 2008/09-2011/12 (436 deaths in 15-44 yr olds and 1,274 in 45-64 yr olds). On average, 76% of seasonal influenza A all-age attributable deaths had a cardiovascular or respiratory cause recorded (average of 5,849 influenza A deaths per season), with nearly a quarter reported for other causes (average of 1,770 influenza A deaths per season), highlighting the importance of all-cause as well as cause-specific estimates. No significant influenza B attributable mortality was detected by season, cause or age group. This analysis forms part of the preparatory work to establish a routine mortality monitoring system ahead of introduction of the UK universal childhood seasonal influenza vaccination programme in 2013/14.
PLoS ONE 01/2013; 8(12):e79360. · 3.73 Impact Factor
[show abstract][hide abstract] ABSTRACT: In order to comply with national and international clinical laboratory accreditation standards (e.g. the ISO 15189, Clinical Pathology Accreditation standards) and with the joint code of practice for research, there must be a method of assessing that test methods are "fit for purpose". This document gives guidance on development and describes how a validation file is produced. A test method may be a commercial kit, an in-house assay or reagent or a set of reagents bought separately and used to prepare an in house assay. A validation file is needed for both current and new test procedures. The file may refer to data recorded in workbooks, papers and reports. Modifications to assays (including commercially available assays) necessitate either an update to the validation file or creation of a new file. This paper is intended to provide a generic framework for in-house assay development and validation of new nucleic acid amplification assays including real-time polymerase chain reaction (PCR).
Journal of clinical virology: the official publication of the Pan American Society for Clinical Virology 12/2012; · 3.12 Impact Factor
[show abstract][hide abstract] ABSTRACT: The intense influenza activity in England during the 2010-11 winter resulted from a combination of factors. Population-based seroepidemiology confirms that the third wave of influenza A(H1N1)pdm09 virus circulation was associated with a shift in age groups affected, with the highest rate of infection in young adults.