Identification of human parvovirus B19 among measles and rubella suspected patients from Cuba

Between September 2014 and December 2015, 298 sera from rash and fever patients from all over Cuba were investigated for specific IgM antibodies against measles, rubella, dengue, human parvovirus B19 (B19V) and human herpesvirus 6 (HHV6) using a commercial enzyme-linked immunosorbent assay kits. B19V IgM positive and equivocal samples were investigated by a polymerase chain reaction and genotyping. No measles, rubella or dengue cases were detected. HHV6-IgM antibodies were confirmed in 5.7% and B19V-IgM antibodies in 10.7% of the patients. A total of 31.3% of the B19V cases were between 5 and 9 years old and 34.4% were 20 years and older. The only B19V sequence obtained belonged to genotype 1a. Diagnosis was established for only 16% of the rash and fever patients, suggesting that other diseases such as Zika or Chikungunya may play a role.     

一起应急接种麻疹疫苗后偶合乙型流感的调查报告

目的为了解应急接种麻疹疫苗后发热病人异常增加的原因，指导疫情控制及疫苗应急接种。方法采用现场流行病学调查方法对粤北农村某小学学生应急接种麻疹疫苗后发热病人异常增加的原因进行调查分析。结果该小学有学生655人，应急接种麻疹疫苗644人，接种后发生发热病人34例。3例除有发热、咳嗽外，均有皮疹，采集病例血清标本，麻疹IgM阳性，风疹IgM阴性；其余31例发热病例中，发热38℃及以上30例，占96．77％；咳嗽22例，占70．97％；头晕17例，占54．84％；头痛15例，占48．39％；鼻塞12例，占38．71％；流涕12例，占38．71％；呕吐5例，占16．13％；乏力17例，占54．84％。31例病例均无皮疹，采集2例病例咽拭子标本，检出乙型流感病毒核酸。结论该校发热病例明显增加是应急接种麻疹疫苗后偶合乙型流感所致。提示春季群体性接种疫苗应避免偶合其它传染病的爆发。     

Investigation of measles and rubella outbreaks in Tamil Nadu, India-2003

The aims of the present study were to confirm measles outbreaks by detection of measles-specific IgM antibodies, isolation of measles virus, and genetic characterization to document the circulating genotypes in Tamil Nadu. Eight outbreaks were reported from six districts of Tamil Nadu, India during the period Jan-Dec 2003. Blood samples were collected for serology, urine, and throat swabs for virus isolation. Genotypic characterization of measles isolates was based on the sequence of the N gene. All the clinically suspected outbreaks (n = 8) were confirmed by serology; six out of the eight as measles and two as combination of measles and rubella highlighting the need to carry out rubella serology on measles-negative samples. Genetic characterization of three isolates obtained revealed one as genotype D4 and two as D8. Measles genotypes D4 and D8 were found to circulate in three districts of Tamil Nadu. It is necessary to be aware of the circulating genotypes within the geographical area. The information would be valuable to evaluate control measures and identify viral transmission and importation.     

2009-2013年菏泽市曹县麻疹病毒感染特征分析

目的 分析曹县麻疹病例实验室检测结果、流行病学和临床特点,为当地预防控制麻疹提供指导.方法 从国家疾病监测信息报告管理系统收集2009-2013年曹县麻疹疫情数据,以及麻疹疑似病例,开展麻疹IgM、风疹IgM检测和分析麻疹病毒分离情况、病例特点.结果 2009-2013年度共报告227例麻疹疑似病例,血清标本采集率72.69％,麻疹IgM阳性率为18.71％,12例咽拭子标本中分离出5株麻疹病毒,为H1a基因型.确诊168例麻疹病例,＜4岁年龄组的构成比为80.95％,幼托儿童、散居儿童是麻疹发病的主要人群,占89.29％;主要临床表现包括发热,超过38.5℃者占86.63％,皮疹首发部位为面部,占72.95％,有口腔黏膜斑者占72.80％,麻疹并发症前三位分别为肺炎、心肌炎、喉炎.结论 经血清学和病毒学证实,曹县当地存在麻疹病毒传播,病例以散发为主,麻疹野病毒为H1基因型,麻疹发病与年龄、性别、职业、季节有关.     

Evaluation of rubella IgM enzyme immunoassays.

BACKGROUND: Rubella virus generally causes a mild fever, rash illness similar in clinical presentation to infections by other viruses including measles and parvovirus B19. Rubella infections in pregnant women in the first trimester carry a high risk of congenital rubella syndrome (CRS) which can result in severe congenital defects in the infants. The goal of rubella immunization programs is therefore to eliminate CRS. The primary test for the laboratory confirmation of rubella is IgM serology. It is therefore important to evaluate currently available commercial rubella IgM immunoassays to ensure high quality rubella diagnostic testing. STUDY DESIGN: In this study, we compared the performance of seven commercial rubella IgM enzyme immunoassays (EIA) (Meddens, Denka Seiken, Behring, Wampole, Captia, Sigma and Abbott Axsym) using well-defined panels of sera from rubella and non-rubella/rash-illness cases. RESULTS: The Meddens, Denka Seiken, Behring and Wampole rubella IgM EIAs all performed similarly for sensitivity (range of 74.1-76.8%) and specificity (range of 93.9-96.1%). Relative to the other assays, the Axsym had a higher sensitivity (78.9%) but lower specificity (86.5%). The Captia assay had the lowest overall sensitivity (66.4%), while the Sigma assay had a lower specificity (85.6%) in relation to the other assays. CONCLUSIONS: In conclusion, the Meddens, Denka Seiken, Behring and Wampole rubella IgM EIAs are comparable in their overall performance with respect to sensitivity and specificity.     

Detection of measles,mumps, and rubella antibodies in saliva using antibody capture radioimmunoassay

Antibody capture radioimmunoassays were developed for detecting virus specific IgM (MAC-RIA) and IgG (GACRIA) to measles, mumps, and rubella and used to investigate saliva as an alternative specimen to serum for diagnosis. Saliva was collected from 63 patients with measles, 19 with mumps, and 150 with rubella, which were all clinically diagnosed and serologically confirmed. Virus specific IgM was detected in 92% of measles, 75% of mumps, and 100% of rubella saliva samples collected during the first week of illness. Between 1 and 5 weeks after onset virus specific IgM was detected in 100% of saliva specimens. After the 5th week the proportion of reactive specimens declined. The specificity of the MACRIA tests was established by testing saliva samples collected from blood donors for measles (88), mumps (88), and rubella IgM (91). All of the saliva specimens tested for measles and rubella specific IgM were unreactive, 1/88 specimens tested for mumps specific IgM contained significant reactivity. Saliva specimens collected from acute cases of MMR were tested in all 3 MACRIAs. A small proportion of saliva samples contained detectable IgM of more than one virus infection. Rubella and measles specific IgG was detected in the saliva of all cases from the 4th or 5th day of illness, respectively. Detection of mumps specific IgG was less successful. We have demonstrated that virus specific IgM can be reliably detected in saliva samples collected from acute cases of measles, mumps, and rubella and identified 1–5 weeks after onset of illness as the optimum time for collection of samples. © 1993 Wiley-Liss, Inc.     

Circulating immune complexes in patients with acute measles and rubella virus infections

A solid-phase C1q radioimmunoassay was used to test for immune complexes (ICs) in sera obtained longitudinally from patients recovering from acute, uncomplicated measles and rubella virus infections. ICs were detected in 12 (18.5%) of 65 sera from 14 measles patients who did not have prolonged IC formation. Of 12 IC-positive measles sera, 9 were collected 4 weeks or more after rash onset. Transient appearance of detectable circulating ICs occurred sooner in 22 rubella patients who did not have prolonged IC formation. Of 109 rubella sera, 14 (12.8%) were IC-positive, and, of these, 10 were collected within 3 weeks of rash onset. Prolonged IC formation was found for an additional four measles and two rubella virus patients. Fractionation of sera from these six patients revealed that levels of large-sized ICs were highest in the initial 10 days after rash onset. Levels of large-sized ICs then declined to those for medium- and small (approximately immunoglobulin G)-sized ICs. IC-associated virus-specific antigens were detected in some of the sera from the six patients having prolonged IC formation. These results suggest two things: first, measles and rubella virus patients differ in the timing of virus clearance or in the reestablishment of normal immunity after infection; second, virus clearance is prolonged in some measles and rubella virus patients who have seemingly normal recoveries from their infections.     

Evaluation of diagnostic markers for measles virus infection in the context of an outbreak in Spain

A measles outbreak occurred from January to July 2003 in Spain, despite the fact that the Plan of Eradication of Measles and its surveillance program had been set up in 2001. Different diagnostic markers for measles virus infection were compared for 246 patients in tests of serum, urine, and pharyngeal exudate specimens. Measles virus immunoglobulin M (IgM) and IgG and rubella virus and parvovirus IgM levels in serum were assayed. Multiplex PCR was done on urine, serum, and pharyngeal exudates, and isolation of measles virus in the B 95 a cell line from urine was attempted. At least one positive marker for measles virus was obtained from 165 patients (67.1%; total number of patients, 246). A total of 136 cases (82.4% of the patients showing positive markers) were diagnosed by PCR and/or isolation and IgM detection methods. The results for 27 patients (16.4%) were positive only by direct methods. The results for two patients (1.2%) were positive only by IgM detection. In the case of the first group (136 cases), the time elapsed from appearance of the rash was significantly longer than in the case of the group which was only positive by PCR. Besides, 8 out of 27 PCR-positive IgM-negative cases showed specific IgG results, suggesting either secondary vaccine failure or reinfection. Numbers resulting from PCR performed with pharyngeal exudates proved to be significantly higher than those obtained with other specimens. Phylogenetic analysis showed the presence of genotype B3. The results strongly back the World Health Organization recommendation that detection of IgM should be supplemented by PCR and isolation for the diagnosis of measles virus infection.     

Development of a measles specific IgM ELISA for use with serum and oral fluid samples using recombinant measles nucleoprotein produced in Saccharomyces cerevisiae.

In order to develop sensitive assays for detecting measles antibodies in oral fluid specimens, we have produced recombinant measles virus nucleoprotein (rMVN) in a yeast expression system and prepared monoclonal antibodies to the protein. Measles nucleoprotein gene from the Schwarz vaccine strain was cloned into a yeast expression vector, pFX7 under the control of the hybrid GAL10-PYK1 promoter. High levels of rMVN (20 mg/litre of yeast culture) were generated. Electron microscopy showed that the purified rMVN assembled into typical herring-bone structures. Monoclonal antibodies produced to the rMVN also reacted with native measles virus N in immunofluorescence tests. The purified rMVN and a monoclonal antibody to the rMVN conjugated to horseradish peroxidase were used to develop a measles specific IgM capture EIA (MACEIA) in both serum and oral fluid specimens. Evaluations of the MACEIA were performed by testing a) serum samples (n=80) and b) paired oral fluid/serum samples from measles cases (n=50, representing 16 cases) and oral fluids from controls with non-measles rash (n=59, representing 48 cases). The samples were also tested for measles IgM, using a reference radioimmunoassay (MACRIA). The sensitivity and specificity of the MACEIA compared with MACRIA for a) the serum samples were 100 and 96.6% respectively and b) for paired serum/oral fluids samples 100 and 100%, respectively.     

Prévalence de l’infection par le parvovirus B19 humain au cours des éruptions fébriles de l’enfant dans le Nord tunisien

The aim of the study is to evaluate the prevalence of specific antibodies anti-human parvovirus B19 (PVB19) immunoglobulin M (IgM) and IgG in children with fever and rash. This study involved 257 children aged from 7 months to 15 years with febrile rash unrelated to measles and rubella (seronegative for IgM). The sera were examined by immunoenzymatic assay. Detection of antibodies of PVB19 was done by enzyme-linked immunosorbent assay (Elisa). In our study, prevalence of immunoglobulin G (IgG) and IgM were 44 and 11.3%, respectively. Clinically, children with positive IgM serology had submitted an erythema infectiosum (13/29 cases), myocarditis (1 case), encephalitis (1 case), severe sickle cell anemia (7 cases), and immunocompromised (7 cases). The incidence rate of viral infection was 11.3%; most of the cases of PVB19 infection occurred between the months of May and August. Incidence was higher in the 10–15 years age group (21%). The prevalence of IgG antibody varied and increased with age, it rises from 38.2% in preschool children (19 months–4 years) to 53.5% in those aged between 4.5 and 15 years, reaching 58% in the 10–15 years age group. The four risk factors of PVB19 infection are: (1) those aged between 4.5 and 9 years, which is the most affected age group (P = 0.0018); (2) female gender in children aged between 19 months and 4 years (P = 0.037); (3) transfusion and (4) immune deficiency (P = 0.022 and P = 0.001, respectively). The study of the prevalence of PVB19 infection shows that viral infection is acquired early in childhood, increases with age; viral transmission is favored by the community life. Because of the widespread vaccination program against measles and rubella, the systematic search of PVB19 in front of eruptive fevers becomes important.     

泉州市2009年麻疹监测结果分析

目的 分析泉州市2009年麻疹监测结果,为制定消除麻疹策略提供科学依据。方法 按国家麻疹监测方案开展监测工作,应用描述流行病学方法和Excel 2003软件对2009年麻疹监测结果进行分析。结果 2009年共有132例麻疹疑似病例纳入麻疹监测信息报告管理系统管理,确诊15例,发病率为0.19/10万,排除病例报告发病率﹥1/10万的县（市、区）占41.67%;48 h个案调查率为96.09%,7 d内血清检测结果及时报告率达到85.51%;血标本采集率为52.27%（69/132）,检测麻疹IgM阳性率为21.74%,检测风疹IgM阳性率为42.03%。73.33%的病例集中在晋江;53.33%的病例集中在1月份和4月份;66.67%的病例集中在0～7月龄小年龄组和﹥14岁的大年龄组;无免疫史和免疫史不详者占73.33%;66.67%的病例是流动人口;60%的病例在发病前7～21 d去过医院。结论 泉州市2009年麻疹监测的敏感性和特异性均较低;无MV免疫史和免疫史不详的人群是麻疹发病的高危人群。     

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.     

Assessment of RNA amplification by multiplex RT‐PCR and IgM detection by indirect and capture ELISAs for the diagnosis of measles and rubella

Sanz JC, Mosquera M, Ramos B, Ramírez R, de Ory F, Echevarria JE. Assessment of RNA amplification by multiplex RT‐PCR and IgM detection by indirect and capture ELISAs for the diagnosis of measles and rubella. APMIS 2010; 118: 203–9. The aim of the study was to compare RNA amplification using multiplex RT‐PCR and IgM detection by means of indirect and capture ELISAs for the diagnosis of measles and rubella. A total of 229 cases of maculopapular rash with serum and throat swab samples were included. Specific serological IgM to measles and rubella was determined by Enzygnost^® (Siemens) and Platelia^TM (Bio‐Rad). Both viruses were researched using multiplex RT‐PCR performed on throat samples. Criteria for inclusion of measles or rubella cases were a positive RT‐PCR result for one virus and negative for the other; and/or a positive IgM result for one virus by both ELISAs and negative RT‐PCR for the other virus. A total of 74 cases were classified as measles and 54 as rubella. In measles, sensitivity and specificity were 93.2% and 100% for RT‐PCR, 97.3% and 98.1% for Enzygnost^®, and 90.5% and 95.5% for Platelia^TM. For rubella, these values were 42.6% and 100% for RT‐PCR, 100% and 97.1% for Enzygnost^®, and 94.4% and 98.3% for Platelia^TM. Enzygnost^® and Platelia^TM are useful techniques for detecting IgM against measles and rubella. RNA amplification by RT‐PCR was both sensitive and specific for the diagnosis of measles; however, for rubella, the sensitivity of this technique must be improved.     

Monitoring progress toward measles elimination by genetic diversity analysis of measles viruses in China 2009–2010

With the achievement of high coverage for routine immunization and supplementary immunization activities (SIAs), measles incidence in mainland China reached its lowest level in 2010. The proportion of measles cases in the vaccination-targeted population decreased during 2007–2010 after the SIAs. More than 60% of measles cases were in adults or infants, especially in the coastal and eastern provinces during 2009 and 2010. A total 567 isolates of measles virus were obtained from clinical specimens from 27 of 31 provinces in mainland China during 2009 and 2010. Except for two vaccine-associated cases, one genotype D4 strain, two genotype D9 strains, and four genotype D11 strains, the other 558 strains were genotype H1 cluster H1a. Genotype H1 has been the only endemic genotype detected in China since surveillance began in 1993. Only genotype H1 was found in mainland China during 1993–2008, except for one detection of genotype H2. More recently, multiple genotypes of imported measles were detected even with the background of endemic genetotype H1 viruses. Analysis of the 450-nucleotide sequencing window of the measles virus N gene showed that the overall genetic diversity of the recent geneotype H1 strains decreased between 2008 and 2010. The lower genetic diversity of H1 strains suggested that enhanced vaccination may have reduced the co-circulating lineages of endemic genotype H1 strains in mainland China.     

Concomitant administration of varicella vaccine with combined measles,mumps, and rubella vaccine in healthy children aged 12 to 24 months of age

The reactogenicity and immunogenicity of three combined measles, mumps and rubella (MMR) vaccines and one administered with a varicella vaccine was studied in infants. The vaccines were Priorix (designated MeMuRu, Group 1), M-M-R II (Group 2), Triviraten (Group 3) and Priorix + a varicella vaccine, Varilrix (Group 4). Fever was greater in Group 2 (61.3%) compared to Group 1 (48.5%; p = 0.033) or Group 3 (37.1%; p = 0.009). Rash with fever was reported in Group 2 (4.8%) and Group 4 (3.3%), but not for Group 1. Anti-measles, -mumps and -rubella seroconversion was similar for Group 1 (96.1%, 96.1% and 100%, respectively), Group 4 (98% for all three), and Group 2 (91.5%, 93.6% and 97.9%) 60 days post-vaccination. GMTs for measles (3,053.7-3,412.2 mIU/ml), mumps (1,001.5-1,158.8 U/ml) and rubella (68.7-89.1 IU/ml) were similar for Groups 1, 2 and 4 at Day 60. Antibody persistence was noted 2 years post-vaccination. The MeMuRu + varicella combination showed no clinically relevant increase in reactogenicity and should facilitate introduction of a varicella vaccine into national immunization schedules.     

Molecular epidemiology of circulating measles virus in Ireland 2002–2007

The molecular characterization of measles virus (MeV) is a valuable epidemiological tool to monitor virus transmission and to discriminate between imported and endemic infection. There has been significant immigration into Ireland in recent years and many individuals originate from regions of high measles incidence. Ireland has had a number of outbreaks of MeV which appear attributable to sub‐optimal vaccine uptake and possibly imported strains as new genotypes have been identified in recent years. To ascertain any significant changes in circulating measles genotypes we investigated 65 confirmed measles cases between the years 2002 and 2007. The laboratory diagnosis of measles was confirmed by detection of measles‐specific IgM in oral fluid in conjunction with a real‐time polymerase chain reaction assay targeting the MeV hemagglutinin gene. Phylogenetic analysis based on the 3′ hypervariable region of the nucleoprotein gene was performed and three genotypes, all within measles clade D, were found to be circulating during this time period. In 2002 and 2003, genotype D8 (n = 2) was observed whereas genotype D7 was dominant in 2003 (n = 31). A distinct change in the circulating MeV genotype and increased genetic diversity was observed between 2004 and 2007. All cases were within genotype D4 (n = 32) but were phylogenetically distinct from each other. These data provide important epidemiologic baseline information on MeV in Ireland and facilitates detailed examination of measles transmission. J. Med. Virol. 81:125–129, 2009. © 2008 Wiley‐Liss, Inc.     

2013—2017年北京市通州区不同剂次水痘疫苗免疫史突破病例特征分析

目的了解2013—2017年北京市通州区不同剂次水痘疫苗免疫史突破病例流行病学及临床特征。方法通过现场调查和查阅中国疾病控制信息系统和北京市免疫规划信息系统获得水痘发病数据和免疫史资料,采用描述流行病学方法进行分析。结果2013—2017年通州区共报告中小学、托幼机构水痘病例数2102例,原发水痘病例989例,有明确1剂次免疫史的水痘突破病例966例,2剂次免疫史的突破病例147例。不同免疫史患者的性别、年龄、职业构成比差异有统计学意义。原发病例中,发热病例占46.71%,中重度出疹病例占34.68%;1剂次突破病例中发热病例占41.20%,中重度出疹病例占17.39%;2剂次水痘突破病例中发热病例占26.53%,中重度出疹比例占7.48%。3组的发热、出疹构成比差异有统计学意义。1剂次突破病例发病间隔中位数为5.11年,2剂次突破病例发病间隔中位数为2.44年,1剂次和2剂次突破病例发病间隔差异有统计学意义。结论随着免疫接种剂次的增加,水痘突破病例发热和出疹症状越来越不典型,由于临床症状相对较轻,突破病例在集体单位中作为传染源更容易被忽视。     

Antibodies against different measles antigens and rubella in patients with multiple sclerosis and other neurological diseases

Antibodies against measles virus have been demonstrated in CSF of some patients with MS or a related disease as optic neuritis or myelopathy. This report describes antibody measurements in CSF of MS patients against different measles antigens and various other virus antigens compared with other nonmyelinating neurological diseases. CSF specimens were obtained from 63 MS patients and from 101 other neurological patients with diagnosis as meningo-encephalitis, Parkinsonism, GuillanBarré and disturbances in BBB. CF antibodies were measured against herpes simplex, zoster-varicella, mumps and RSSE, and HI antibodies against measles and rubella. Additional tests were measles HLI, RNP gel-precipitation, and quantitation of total proteins, IgG and albumin. No CF antibodies were found in either group. Positive rates of measles tests im MS patients were from 20 to 60%, and 6 to 9% in controls. Six control patients showed antibodies in measles HLI test. In a followup study a demyelinative process or disturbance in BBB was evident in four cases. The concentration of IgG was significantly higher among MS patients, but not amounts of albumin or total proteins, suggesting intrathecal synthesis of IgG. Rubella HI antibodies were found in 18% of MS patients compared with 2% in control group. Also rubella antibodies seemed to be correlated with a higher IgG level.