Isolation and genotyping of rubella virus from a case of congenital infection in Brazil

The incidence of CRS and CRI has decreased markedly worldwide with the implementation of efficient vaccination programs. We report a congenital rubella case with fetal death occurred at 29th week of gestation. RV was confirmed in placenta. The results of phylogenetic analysis showed that the RVs/SaoPaulo01.- BRA/08.CRI belongs to the genotype 2B of RV.     

Application of new assays for rapid confirmation and genotyping of isolates of rubella virus

Rubella virus (RV) isolation is recommended by the WHO Measles and Rubella Labnet for studying the etiology and epidemiology of rubella. However, the absence of cytopathologic effects (CPE) in many of the cell lines used commonly makes it difficult to confirm RV growth. In this study, two assays amplifying RV cDNA were developed and validated in order to confirm and genotype RV isolates after cell culture. A SYBR Green I-based real-time PCR (Rtime-SGE317) was established for initial rapid detection of RV in Vero cells and a nested PCR (PCR-E860) was used for amplifying further the 739 nt window of the E1 gene for the identification of RV genotype as recommended by the WHO. Sensitivities of the two assays were evaluated using eight RV isolates, two from infants with the congenital rubella syndrome (CRS) and six from patients with acute rubella. All the isolates had cycle threshold (C(t)) values <37 after the third passage, which is recommended as the cut-off for the confirmation of a viable RV isolate. Phylogenetic analysis based on the 739 nt window generated by the PCR-E860 showed that the eight RV isolates belonged to genotypes 1E, 1G, and 2B. The Rtime-SGE317 assay can be carried out in local public health laboratories, which would extend the molecular surveillance of rubella and contribute to the WHO goal of eradicating rubella worldwide.     

Epidemiological and molecular assessment of a rubella outbreak in North‐Eastern Italy

From January to June 2008, a rubella outbreak involving 111 laboratory confirmed cases occurred in the Friuli Venezia Giulia (FVG) region of North‐Eastern Italy. The outbreak occurred initially in two residential homes for young adults disabled mentally and physically. Subsequently, the epidemic spread to the general population. Young adult cohorts were mostly affected and the mean age of the patients was 26.8 years; the majority of cases were male (73.8%), with a mean age of 26.6 years in males and 27.4 in females. Three pregnant women had a primary infection and two had their pregnancies terminated. Genotyping of 16 isolates showed the circulation of RUBV 2B, a genotype originating from Asia and South Africa and now present in Europe. In addition, molecular analysis revealed a well defined space‐temporal spread of two viruses showing distinct sequences. A seroepidemiological survey carried out in a city within the same geographical area showed that the proportion of women of childbearing age still susceptible to rubella virus was 5.5%, fairly close to the figure (<5%) expected by 2010. J. Med. Virol. 82:1976–1982, 2010. © 2010 Wiley‐Liss, Inc.     

2010-2012年广州地区人冠状病毒及其亚型分布

目的了解2010—2012年广州地区发热呼吸道感染病人的冠状病毒（HCoV）流行状况和特点,为进一步了解人冠状病毒的流行规律及防控奠定基础。方法收集广州市2010—2012年5家哨点医院共3376例符合要求的发热同时伴随有呼吸道感染症状病人的咽拭子标本及相关临床信息,采用Real—timePCR方法检测冠状病毒的核酸并对其进行主要型别鉴定,从人群分布、月份分布及各型别发病率等方面进行分析。结果3376例发热伴有呼吸道感染症状的病例中,55例为冠状病毒阳性标本,检出率为1．63％。亚型0C43、229E和NL63阳性的标本分别为23、13和19例,阳性率分别为0．68％、0．39％和0．56％,其中SARS、HUKl及EMC亚型未检出。冠状病毒流行高峰为1月（5．33％）,不同月份之间发病率比较差异有统计学意义（xz=38．197,P〈0．01）;阳性率最高年龄组为60。93岁,检出率为2．6％。亚型OC43发病高峰为12—1月,与其他月份比较差异有统计学意义（xL30．054,P〈0．01）,检出率最高的年龄组为60～93岁（1．3％）;229E月分布较为分散,各月份间差异无统计学意义（x：=15．651,P〉0．05）,在各年龄组的分布呈不集中趋势;NL63流行高峰为6-9月,与其他月份比较差异有统计学意义（x2=5．801,P〈0．05）,检出率最高的年龄组为7～17岁（1．03％）。结论广州市2010—2012年冠状病毒流行高峰为1月份。老年组检出率最高;亚型中OC43阳性率最高,NL63次之,229E亚型阳性率最低;0C43流行高峰为冬季,老年组检出率较高;229E流行高峰及感染人群较为分散;NL63流行高峰为夏季,儿童青少年组检出率最高。     

Epidemiology of influenza in hospitalized children with respiratory tract infection in Suzhou area from 2016 to 2019

Influenza is a contagious respiratory disease and risks public health in China, and it has caused wide public concern in recent years. Immunocompromised patients, such as children and elderly people, suffer more severe influenza complication and some extreme cases are even life threatening. To identify the influenza characteristics and its correlation with various climatic and environmental pollution factors, we collected the reported influenza epidemic of hospitalized children in Children's Hospital of Soochow University from 2016 to 2019. Our results show that the main influenza virus subtypes are A/H1N1, A/H3N2, B/BV, and B/BY. We also identified the characteristics of the prevalent influenza virus subtypes in different months, seasons, years, and patients' age. Of all the influenza infected patients, the most susceptible groups are children over 3 to 5 years of age, and more cases are reported in winter than other seasons. We also found that influenza is also highly correlated with climatic and environmental pollution factors, and the autoregressive integrated moving average model is employed for the short-term influenza prediction in Suzhou city, which can provide scientific basis for the prevention and control of influenza and public health decision-making.     

Molecular epidemiology of rubella viruses involved in congenital rubella infections in São Paulo,Brazil, between 1996 and 2009

Rubella virus (RV) infection during the early stages of pregnancy can lead to serious birth defects, known as congenital rubella syndrome (CRS). This retrospective study was conducted between 1996 and 2009 with surveillance specimens collected from patients suspected of congenital rubella infection (CRI) and CRS. The clinical samples (nine amminiotic fluid, eight urine, eight blood, one conception product, and one placenta) were sent for viral isolation and genotyping. Twenty‐seven sequences were analysed and four genotypes (1a, 1B, 1G, and 2B) were identified in São Paulo that were involved in congenital infection. To our knowledge, this study is the first report that describes genetic diversity of the circulating rubella strains involved in CRI. J Med. Virol. 85:2034–2041, 2013. © 2013 Wiley Periodicals, Inc.

     

孕前及孕期妇女风疹免疫力调查和风疹近期感染检测

目的了解孕前妇女及孕期妇女风疹易感水平和近期感染状况，为预防先天风疹感染做好优生优育工作提供依据。方法用生物蛋白芯片技术对2986例孕前妇女及863例孕期妇女（包括725例正常妊娠和138例异常妊娠）进行风疹特异性抗体IgM、IgG检测。结果80．27％的孕前妇女具有免疫力，4．99％孕前妇女为风疹近期感染，14．03％妇女对风疹易感。89．00％的孕期妇女具有免疫力，1．96％孕期妇女为风疹近期感染，7．76％妇女对风疹易感。孕前妇女及孕期妇女的风疹近期感染率和易感率比较有显著性差异（χ^2=14．797、23．846，P〈0．001）；不良妊娠与正常妊娠之间风疹近期感染率和易感率比较有显著性差异（X。=30．635、4．174，P〈0．05）；市区与农村的孕前妇女及孕期妇女在风疹近期感染率和易感率比较无明显不同；孕前妇女及孕期妇女风疹近期感染月份分布均以5月最高。结论本地区孕前妇女风疹感染率高；风疹感染与不良妊娠密切相关：市区与农村妇女的风疹易感水平和近期感染状况相当。对孕前妇女及孕期妇女同时检测风疹特异性抗体IgM、IgG可以正确判定妇女的免疫状况，有效预防、主动发现并合理处理孕妇感染，从而降低风疹感染率提高优生优育水平。     

High prevalence of hepatitis B virus genotype E in Northern Madagascar indicates a West‐African lineage

The prevalence of hepatitis B virus (HBV) markers was investigated in 563 inhabitants aged 15–55 years from a sugar cane region, Sirama, and from a village, Mataipako, in Northern Madagascar. Serological markers of past or present infection were significantly higher in Sirama, 74% versus 45%. There was no difference in the prevalence of chronic HBsAg carriers, 8.7% versus 8.5% between the two regions. Sequencing the S gene in 45 strains revealed a predominance of genotype E, in 53%, followed by subgenotype A1 in 22%, and genotype D in 18%. Phylogenetic analyses of the genotype E strains showed homology with West African strains. All A1 isolates were similar to Malawi strains. Most genotype D strains were subgenotype D7 and related to strains from Somalia and Tunisia. One genotype D strain formed a branch between Pacific D4 and African D7 strains at neighbor‐joining analysis. The pre‐core stop mutant was found in 33% of the genotype D strains, 17% of E but not in any A1 strain. The high prevalence and low variability of genotype E strains in only two villages, indicates a rather recent introduction of this genotype into Madagascar from West Africa, possibly through migration or slave trade. The wider spread and genetic relationship of genotype D with East African and Austronesian strains indicate an earlier introduction of this genotype. Molecular epidemiology of HBV may thus be used to complement linguistic and genetic studies on past human migrations in Africa. J. Med. Virol. 82:1515–1526, 2010. © 2010 Wiley‐Liss, Inc.     

Epidemiological and molecular characterization of rubella virus isolated in São Paulo,Brazil during 1997–2004

Rubella virus (RV) infection during the early stages of pregnancy can lead to serious birth defects, known as the congenital rubella syndrome (CRS). In 2003, the Pan American Health Organization (PAHO) adopted a resolution calling for the elimination of rubella and the congenital rubella syndrome (CRS) in the Americas by the year 2010. Brazil will have implemented the recommended PAHO strategy for elimination and interruption of endemic rubella virus transmission. The characterization of genotypes during the final stages of rubella elimination is important for determining whether new rubella isolates represent endemic transmission or importations. Samples (blood, urine, cerebrospinal fluid, and throat swabs) collected from patients with symptoms suggestive of rubella infection in 1997–2004 were isolated in cell culture and genotyped. Twenty‐eight sequences were analyzed and two genotypes were identified: 1a and 1G. The information reported in this paper will contribute to understanding the molecular epidemiology of RV in São Paulo, Brazil. J. Med. Virol. 84:1831–1838, 2012. © 2012 Wiley Periodicals, Inc.     

Emergence and diversity of different HIV‐1 subtypes in South Africa, 2000–2001

HIV‐1 is a major health problem in South Africa with an average prevalence rate of 29.1% in pregnant women and between 4.9 and 6.1 million people infected. Using env gp120 V3 serotyping and genotyping techniques 410 patient samples were investigated. Most of the samples were obtained from different clinics in the greater Cape Town area of the Western Cape Province in South Africa. These included an academic hospital, state and private clinics, an informal settlement, sex worker cohorts, and the blood transfusion services. RNA was extracted from plasma samples followed by RT‐PCR and sequencing of the env gp120 V3 region. Sequence fragments were assembled using Sequencher V4.7 and subsequently codon aligned. Distance calculation, tree construction methods, and bootstrap analysis were implemented using MEGA version 4.0. Viral load measurements indicated that HIV‐1 RNA levels from 74 samples were below the assay detection limit. Three hundred thirty‐six samples were used for env PCR and sequencing and 320 were assigned to subtypes. The majority of the sequences were subtyped as C (n = 285, 89.0%). Other subtypes detected were subtype A (n = 10, 3.1%); subtype B (n = 22, 6.8%); one each of subtypes F1, G, U, and a CH recombinant. Whether this diversity will have major implications for HIV‐1 evolution and vaccine development in this region remains undetermined. J. Med. Virol. 81:1852–1859, 2009. © 2009 Wiley‐Liss, Inc.     

Synthesis of Saccharide‐Terminated Poly(ε‐caprolactone) via Michael Addition and ‘Click’ Chemistry

Maleimido‐terminated PCL (M‐PCL) and alkyne‐terminated PCL (A‐PCL) are prepared by the ring‐opening polymerization of ε‐caprolactone with N‐hydroxyethyl maleimide and 4‐pentyn‐1‐ol as initiators catalyzed by tin(II ) trifluoromethane sulfonate at 25 °C, respectively. A series of saccharide‐terminated PCLs have also been synthesized under mild conditions by two chemical strategies: 1). Michael addition of M‐PCL and amino‐containing maltose, and 2). a ‘click’ reaction of A‐PCL and azide‐containing saccharide. The amphiphilic nature of these maltose‐terminated PCLs make self‐assembly into aggregates in water possible. These aggregates have been characterized by transmission electron microscopy and dynamic light scattering measurements.

     

Comparison of various serological methods and diagnostic kits for the detection of acute, recent, and previous rubella infection, vaccination, and congenital infections

The antibody development after natural rubella infection and rubella vaccination has been followed in 802 sera from 493 patients and 71 sera from 22 vaccinees. Also examined were 67 sera from 28 infants with rubella embryopathy and sera from 50 children with presumed prenatal infection. In addition, 777 sera from 641 patients tested for routine rubella diagnosis were studied. Anamnestic information was available from all these patients. These sera were assayed for IgM antibody detection by sucrose density gradient (SDG), the commercial ELISAs (Enzygnost IgM and Rubazyme M), and the non-commercial anti-my-hemadsorption immunosorbent technique (HIT). For the determination of IgG antibodies the hemagglutination inhibition test (HAI), the commercial ELISAs (Enzygnost IgG, Rubazyme), and a single radial hemolysis test (SRH) were used. The SDG and HIT were less sensitive for IgM antibody detection than the two ELISAs, particularly when IgM concentrations were low. In total 26.5% of the IgM results with the newer tests were discordant with SDG, but only 0.5-1.3% of these results were not explicable when the clinical data was considered. Problems were encountered with all IgM assay systems used. For the detection of rubella antibodies after acute infection and vaccination the ELISA Enzygnost IgG was as sensitive as the HAI whereas the ELISA Rubazyme and SRH detected antibodies with some delay. Corresponding results with all tests were found more than 25 days after acute infection and more than 50 days after vaccination. All methods can be used for detection of antibodies in infants with rubella embryopathy. The results of this study suggest that certain combinations of tests can be used for the reliable detection of rubella infection.     

99th Dahlem Conference on Infection,Inflammation and Chronic Inflammatory Disorders: Darwinian medicine and the ‘hygiene’ or ‘old friends’ hypothesis

The current synthesis of the ‘hygiene hypothesis’ suggests that the recent increase in chronic inflammatory disorders is at least partly attributable to immunodysregulation resulting from lack of exposure to microorganisms that have evolved an essential role in the establishment of the immune system. This document provides a background for discussion of the following propositions.

• 1 The essential role of these organisms is an example of ‘evolved dependence’.
• 2 The most relevant organisms are those that co‐evolved with mammals, and already accompanied early hominids in the Paleolithic.
• 3 More recently evolved ‘childhood infections’ are not likely to have evolved this role, and recent epidemiology supports this contention.
• 4 This mechanism is interacting with other modern environmental changes that also lead to enhanced inflammatory responses [inappropriate diet, obesity, psychological stress, vitamin D deficiency, pollution (dioxins), etc.].
• 5 The range of chronic inflammatory disorders that is affected is potentially larger than usually assumed [allergies, autoimmunity, inflammatory bowel disease, but also vascular disease, some cancers, depression/anxiety (when accompanied by raised inflammatory cytokines), and perhaps neurodegenerative disorders and type 2 diabetes].

     

Genetic suppression of collapsin response mediator protein 2 phosphorylation improves outcome in methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine‐induced Parkinson’s model mice

Parkinson's disease (PD) is a common neurodegenerative disorder characterized by slow and progressive degeneration of dopaminergic neurons in the substantia nigra pars compacta (SNc). Levodopa (l ‐Dopa), the current main treatment for PD, supplies dopamine, but it does not prevent neurodegeneration. There is thus no promising remedy for PD. Recent in vitro study showed the increase in the phosphorylation levels of Collapsin Response Mediator Protein 2 (CRMP2) is involved in dopaminergic axon degeneration. In the present study, we report elevation of CRMP2 phosphorylation in dopaminergic neurons in SNc after challenge with the dopaminergic neurotoxin 1‐methyl‐4‐phenyl‐1,2,3,6‐tetrahydropyridine (MPTP), a common model for PD. Genetic suppression of CRMP2 phosphorylation by mutation of the obligatory Cyclin‐dependent kinase 5 (Cdk5)‐targeted serine‐522 site prevented axonal degradation in the nigrostriatal pathway of transgenic mice. As a result, the degree of MPTP‐induced motor impairment in the rotarod test was suppressed. These results suggest that suppression of CRMP2 phosphorylation may be a novel therapeutic target for PD.     

Co‐circulation of multiple measles virus genotypes during an epidemic in France in 2008

In 2008, measles reappeared in France in a series of outbreaks. During this period, 604 measles cases were reported to a routine surveillance system and 305 (50%) of these cases were then confirmed in the laboratory. To understand better the current epidemiological situation and the circulation of different measles strains, a phylogenetic characterization of 113 (19%) of the measles cases from these outbreaks was performed. All measles cases met the WHO clinical criteria and were confirmed either by laboratory detection of measles‐specific IgM and/or by detection of the virus genome by polymerase chain reaction (PCR) and viral isolation. PCR products generated from blood, oral fluid, urine, or nasopharyngeal‐swab samples were sequenced for molecular epidemiology studies. Phylogenetic analysis showed a co‐circulation of genotypes D4 and D5 during the first measles outbreak in the city of Reims in early 2008. Over the course of the year, the A, B3.2, D8, and D9 genotypes also appeared. The data from this study show the simultaneous circulation of several measles genotypes in France and describe genotypes D8 and D9 for the first time in this country. The data also suggest that there are still many pockets of unvaccinated individuals helping to maintain the circulation of measles virus in the population. Phylogenetic studies allowed the corroboration of epidemiologic links and showed that nosocomial transmission can create significant risk for measles dissemination. Finally, the pattern of changes in viral genotypes during 2008 suggests a regular introduction of measles strains from abroad. J. Med. Virol. 82:1033–1043, 2010. © 2010 Wiley‐Liss, Inc.