Mumps Virus Genotype Research Articles

Genomic diversity of mumps virus and global distribution of the 12 genotypes.

The WHO recently proposed an updated nomenclature for mumps virus (MuV). WHO currently recognizes 12 genotypes of MuV, assigned letters from A to N (excluding E and M), which are based on the nucleotide sequences of small hydrophobic (SH) and haemagglutinin-neuraminidase (HN) genes. A total of 66 MuV genomes are available in GenBank, representing eight of the 12 genotypes. To complete this dataset, whole genomes of seven isolates representing six genotypes (D, H, I, J, K and L) and one unclassified strain were sequenced. SH and HN genes of other representative strains were also sequenced. The degree of genetic divergence, predicted amino acid substitutions in the HN and fusion (F) proteins and geographic distributions of MuV strains were analysed based on the updated dataset. Nucleotide heterogeneity between genotypes reached 20% within the SH gene, with a maximum of 9% within the HN gene. The geographic and chronologic distributions of the 12 genotypes were summarised. This review contributes to our understanding of strain diversity for wild type MuV, and the results support the current WHO nomenclature.

Complete genome sequence of mumps virus genotype g from a vaccinated child in franceville, southeastern Gabon, in 2013.

The genome of mumps virus (MuV), a member of the family Paramyxoviridae of the genus Rubulavirus, consists of a single-stranded, negative-sense, nonsegmented RNA. Here, we report the first whole-genome sequence of 15,263 nucleotides of a mumps virus strain from a 6-year-old vaccinated boy in Franceville, southeastern Gabon.

Anti-mumps virus activity by extracts of Mimosa pudica, a unique Indian medicinal plant

Mumps is an acute and self-limiting disease characterized by parotitis, however in some cases it leads to aseptic meningitis, deafness, encephalitis and orchitis, which is a serious health concern. MMR vaccination was successful in eradicating the disease however, recent reports question the efficacy of MMR vaccine and countless outbreaks are observed in vaccinated populations throughout the world. Lack of specific treatment methods for mumps infection and inefficiency of MMR vaccine in vaccinated populations accentuates the need for the development of novel drugs to control mumps virus mediated serious infections. It was with this backdrop of information that the anti-mumps virus activity of Mimosa pudica was evaluated. Suspected mumps cases were collected to isolate a standard mumps virus by systematic laboratory testing which included IgM antibody assays, virus isolation, RT-PCR and phylogenetic analysis. The virus was quantified by TCID50 assay and anti-mumps virus property was evaluated by CPE reduction assay and cytotoxicity of the extract was measured by MTT assay and phytochemical analysis was done by gas chromatography-mass spectroscopy. The RT-PCR and phylogenetic tree analysis of the SH gene sequence of the clinical isolate showed it to be mumps virus genotype C. 150μg/ml concentration of M. pudica completely inhibited mumps virus and the drug was found to be non-toxic up to 2mg/ml. M. pudica was thus found to be a potent inhibitor of MuV.

Genotyping and subtyping of mumps virus isolates from the Indian subcontinent

Mumps is a vaccine-preventable disease that usually occurs as a self-limiting parotitis, but it can also lead to several life-threatening complications, including pancreatitis, meningitis, and encephalitis. The molecular epidemiology of the virus is poorly understood. The present study describes an outbreak of mumps virus infection in Punjab, India. The etiology was confirmed by serology and RNA detection to be mumps virus in 72 % of the cases and 50 % of contacts. This study, for the first time, revealed the mumps virus genotypes circulating in the Indian subcontinent as subtype G2 of genotype G.

Circulation of Two Mumps Virus Genotypes in an Unimmunized Population in India

Two separate outbreaks of fever with parotitis were reported from the Apsinga and Pimpla villages in the Osmanabad district of the Maharashtra State, India during February and March 2012. Meningo-encephalitis was noted in two patients resulting in the death of an 11-year male. Samples of blood and throat swabs were collected from patients with fever and parotitis. Serum samples from suspected (n=62) and convalescent (n=19) patients were tested for mumps virus specific IgM and/or IgG antibodies. Mumps virus specific IgM antibodies were detected in 44 of 62 serum samples (71%). Of the 19 convalescent phase sera 16 had both, anti-mumps virus IgM and IgG antibodies. Twenty-eight throat swabs collected from patients with parotitis were tested by RT-PCR for the SH gene. Twenty-three specimens were found to be positive and nucleotide sequencing of the amplified PCR products revealed circulation of two distinct genotypes that were village specific. Mumps virus genotype C (n=18) was detected in Apsinga village and genotype G (n=5) in Pimpla village. Two mumps virus isolates were also obtained using Vero cells. This is the first report from India confirming simultaneous circulation of mumps virus genotype C in one village and the G genotype in another village only 37 km away.

Virological Analysis of a Regional Mumps Outbreak in the Northern Island of Japan—Mumps Virus Genotyping and Clinical Description

Virological Analysis of a Regional Mumps Outbreak in the Northern Island of Japan—Mumps Virus Genotyping and Clinical Description

Mumps virus diagnosis and genotyping using a novel single RT-PCR

BackgroundIgM detection is considered as the gold standard for mumps diagnosis. Currently, most cases in developed countries occur in highly vaccinated populations due to secondary vaccine failure. In these patients, pre-existing vaccine-induced antibodies are not able to neutralise the virus, but prevent the typical primary response, so that specific IgM is not always elicited. Consequently, acute infection has to be demonstrated by direct detection of the virus by viral isolation or genomic amplification. RT-PCR allows a diagnosis with the maximum sensitivity to be made and also forms the basis for genotype characterisation by sequencing the SH gene, according to WHO recommendations. However, none of the RT-PCR techniques properly evaluated for the diagnosis of acute mumps infection yields an amplification fragment useful for genotyping, and none of the amplification techniques described for genotyping has proved to be sensitive enough for diagnosis. ObjectivesDevelopment of a RT-PCR for the mumps virus diagnosis and genotyping, properly evaluated in comparison with serological gold-standard technique. Study design195 suspected mumps cases and six wild type MuV genotypes were studied. ResultsOur method was able to detect 0.001 TCID50 of mumps virus. Fifty-eight of these showed positive results, of which 54 (93.3%) showed mumps RNA in saliva, while only 20 (34.5%) had mumps IgM in serum. Genotypes G1, G2, H1, H2, D1 and C were identified in positive samples. ConclusionsThe technique described could be a very useful tool for mumps surveillance, management and control.

Rescue of wild-type mumps virus from a strain associated with recent outbreaks helps to define the role of the SH ORF in the pathogenesis of mumps virus

Mumps virus (MuV) causes acute infections in humans. In recent years, MuV has caused epidemics among highly vaccinated populations. The largest outbreak in the U.S. in the past 20 years occurred in 2005–2006 with over 5000 reported cases in which the majority of the cases was in vaccinated young adults. We sequenced the complete genome of a representative strain from the epidemic (MuV-IA). MuV-IA is a member of genotype G, the same genotype of MuV that was associated with the outbreak in the UK in 2004–2005. We constructed a reverse genetics system for MuV-IA (rMuV-IA), and rescued a virus lacking the open reading frame (ORF) of the SH gene (rMuV∆SH). rMuV∆SH infection in L929 cells induced increased NF-κB activation, TNF-α production and apoptosis compared to rMuV-IA. rMuV∆SH was attenuated in an animal model. These results indicated that the SH ORF of MuV plays a significant role in interfering with TNF-α signaling and viral pathogenesis during virus infection.

Genetic characteristics on the small hydrophobic protein and hemagglutinin-neuraminidase genes of mumps virus in Yunnan province, China from 2007 to 2009

To analyze genetic characterization of the small hydrophobic and hemagglutinin-neuraminidase genes of mumps virus (MuV) isolated in Yunnan province, China from 2007 to 2009. Fourteen MuV strains were isolated in Yunnan, China from 2007 to 2009. Using RT-PCR, the SH gene fragments contained 316 nucleotides in all strains and HN gene of six strains were sequenced. The sequences were aligned with other mumps virus sequences downloaded from GenBank using Mega 4.1 software. Fourteen isolated strains were closely related to other reference strains of F genotypes. In SH gene, the homology of nucleotide and amino acid among the fourteen isolated strains were 98.3% - 100.0% and 96.5% - 100.0%, respectively, and 92.6% - 99.4% and 87.7% - 100.0% of homology when compared with that of strains isolated from other provinces in China, respectively. Wsh1 and Wsh2 strains had less homology when compared to other strains of F genotypes. The fourteen strains had homology of 84.5% - 85.1% and 77.2% compared to vaccine strains on nucleotide and amino acid, respectively, and had homology of 83.4% - 90.9% and 70.1% - 86.0% compared to that of other genotypes. In HN gene, the homology of nucleotide and amino acid among the six isolated strains were 99.3% - 99.5% and 99.1% - 99.7%, respectively, and also 99.8% and 99.8% of homology respectively when compared to the SP strain in China. All the six strains had homology of 92.4% - 93.2% and 95.5% - 96.4% when compared to the vaccine strains on nucleotide and amino acid, respectively, and had homology of 94.7% - 96.8% and 95.5% - 99.1% compared to other genotypes. Fourteen strains isolated in Yunnan from 2007 to 2009 belonged to F genotype of MuV while the HN gene seemed more conservative than SH gene.

Assessment of mumps virus-specific antibodies by different serological assays: which test correlates best with mumps immunity?

Mumps is one of the vaccine-preventable childhood diseases and it has not yet been eradicated in Germany. This raises the question as to whether the available mumps vaccines are effective enough to prevent mumps and which antibody test system allows the authentic assigning of mumps-specific immunity. In an attempt to answer this question, we analysed 227 sera samples from medical students of the University Hospital Frankfurt/Main, Germany, using different test systems: indirect immune fluorescence, neutralisation assay, routine ELISA and newly developed immunoassays, which contain the mumps nucleoprotein and the wild-type strain Enders ATCC VR106, respectively. Mumps vaccination coverage of the screened collective amounted to 75.1%, which differs notably from the detected mumps-specific seropositivity rates in the literature (range 53.3% to 82.4%). In contrast, a small group of unvaccinated students had much higher seropositivity rates. Of course, assigned vaccination coverage and calculated seropositivity rates are not effective enough to interrupt the transmission of the mumps virus. The often-occurring mumps outbreaks, some in highly vaccinated populations, may not always demonstrate vaccine failure. The investigation of newly developed test systems and the occurrence of different mumps virus genotypes should also be considered.

Characterization of Mumps Viruses Circulating in Mongolia: Identification of a Novel Cluster of Genotype H

Although mumps virus is still causing annual epidemics in Mongolia, very few epidemiological and virological data have been reported. We describe here the first phylogenetic analysis data on the mumps viruses circulated in Mongolia in 2009. We detected 21 mumps virus cDNAs and obtained a virus isolate from 32 throat swabs of mumps patients in Ulaanbaatar, the capital of Mongolia. The phylogenetic analyses based on the 316 nucleotides of the small hydrophobic gene show that these sequences form a single cluster, with the closest relatedness to the viruses belonging to genotype H. According to the recommendation of the World Health Organization, Mongolian mumps viruses could be classified into a novel genotype because the divergence between new sequences and genotype H reference viruses is >5% (6.3 to 8.2%). However, additional analyses based on the fusion gene, the hemagglutinin-neuraminidase gene, and the whole-genome indicate that the divergences between the Mongolian isolate and other genotype H strains never exceed the within-genotype divergences of other genotypes. These results suggest that Mongolia strains should be included in genotype H and that the current criteria for mumps virus genotyping should be revised. We propose here that the Mongolian viruses should be classified as a new subgenotype termed H3. Since previous epidemiological studies suggested that genotypes H may be associated with central nervous system diseases, we evaluated the neurovirulence of the Mongolian isolate in the neonatal rat system. However, the virus does not exhibit prominent neurovirulence in rats.

Identification and development of a promising novel mumps vaccine candidate strain

Mumps epidemics are usually caused by airborne transmission of mumps virus (MuV) and have high morbidity in non-immunized children. Epidemiological studies in many regions of China show that the genotype F viral strain is the most prevalent. However, the genotype A strain is currently used to prepare vaccines. Regional epidemiological MuV data suggest a significant application for the development of live attenuated mumps vaccines targeting specific genotypes. This article reports the isolation and culture of a genotype F MuV candidate strain that could be used to prepare a live attenuated mumps vaccine. This strain is shown to have good immunological efficacy and stability in neurovirulence evaluations. This work should facilitate the implementation of mumps vaccination in mainland China by targeting the most prevalent MuV genotype, genotype F.

A 176 amino acid polypeptide derived from the mumps virus HN ectodomain shows immunological and biological properties similar to the HN protein

BackgroundThe hemagglutinin-neuraminidase (HN) protein is the major antigenic determinant of the Mumps virus (MuV) and plays an important role in the viral infectious cycle through its hemagglutination/hemadsorption (HA/HD) and neuraminidase (NA) activities. Objective: analyze the biological and immunological properties of a polypeptide derived from a highly conserved region of the HN ectodomain. Methods: a highly conserved region of the HN gene among several MuV genotypes was chosen to be cloned in a eukaryotic expression vector. The pcDNAHN176-construct was transfected into Vero cells and RNA expression was detected by RT-PCR, while the corresponding polypeptide was detected by immunofluorescence and immunochemistry techniques. The HD and NA activities were also measured. The immunogenic properties of the construct were evaluated using two systems: rabbit immunization to obtain sera for detection of the HN protein and neutralization of MuV infection, and hamster immunization to evaluate protection against MuV infection.ResultsA 567 nucleotide region from the HN gene was amplified and cloned into the plasmid pcDNA3.1. Vero cells transfected with the construct expressed a polypeptide that was recognized by a MuV-hyperimmune serum. The construct-transfected cells showed HD and NA activities. Sera from immunized rabbits in vitro neutralized two different MuV genotypes and also detected both the HN protein and the HN176 polypeptide by western blot. Hamsters immunized with the pcDNAHN176-construct and challenged with MuV showed a mild viral infection in comparison to non-immunized animals, and Th1 and Th2 cytokines were detected in them.ConclusionsThe pcDNAHN176-construct was capable of expressing a polypeptide in Vero cells that was identified by a hyperimmune serum anti Mumps virus, and these cells showed the HD and NA activities of the complete MuV HN protein. The construct also elicited a specific immune response against MuV infection in hamsters.

Mumps Outbreak in the Republic of Moldova, 2007–2008

Moldova experienced a nationwide mumps outbreak between 2007 and 2008. Single-dose monovalent mumps vaccination at 15 to 18 months was introduced in 1983, replaced by a 2-dose MMR schedule at age 1 and 6 to 7 years in 2002. We investigated the outbreak to quantify its extent, explore the role of primary and secondary vaccine failure, and provide control recommendations. We analyzed national mumps surveillance and vaccination coverage data to estimate vaccine effectiveness (VE) using the screening method. A retrospective cohort study in 5 educational institutions was conducted to determine age-specific attack rates (ARs) and VE. We compared vaccine strain-specific ARs. Isolation and genotyping of mumps virus strains were performed. Of 31,142 cases reported during October 2007 and July 2008, 80% were in 15- to 24-year-olds. Of cases with information (66%), 92% were vaccinated once, 4% twice. One-dose mumps VE estimates based on surveillance data over 1997-2001 declined from 91% (95% CI: 88%-92%) in 2-year-olds to 72% (70%-74%) in 15- to 19-year-olds. In the cohort study (n = 1589), VE was -40% (-120% to 20%) for 1 dose. For 2 doses it was 62% (-43% to 90%) in 13- to 15-year-olds. ARs were higher in individuals vaccinated with Urabe strains (43%) than with Leningrad-Zagreb strains (14%, P < 0.001). Mumps virus genotype G5 was identified. Low effectiveness of single-dose mumps vaccination was the main cause of the outbreak. Waning immunity may have contributed to this. The risk of mumps in 2-dose vaccinees was low. Other countries in which large population groups have received <2 doses of mumps vaccine may face similar outbreaks.

Circulation of Mumps Virus Genotypes in Spain from 1996 to 2007

Although the WHO recommends the use of genotyping as a tool for epidemiological surveillance for mumps, limited data on mumps virus (MV) genotype circulation that may be used to trace the patterns of virus spread are available. We describe the first complete series of data from Spain. The small hydrophobic region was sequenced from 237 MV-positive samples from several regions of Spain collected between 1996 and 2007. Six different genotypes were identified: A, C, D (D1), G (G1, G2), H (H1, H2), and J. Genotype H1 was predominant during the epidemic that occurred from 1999 to 2003 but was replaced by genotype G1 as the dominant genotype in the epidemic that occurred from 2005 to 2007. The same genotype G1 strain caused concomitant outbreaks in different parts of the world (the United States, Canada, and the United Kingdom). The remaining genotypes (genotypes A, C, D, and J) appeared in sporadic cases or small limited outbreaks. This pattern of circulation seems to reflect continuous viral circulation at the national level, despite the high rates of vaccine coverage.

Genotyping of mumps virus circulating in Turkey in the 2006–2007 winter season

In the winter of 2006-2007, several parotitis cases were reported in different provinces of Turkey. Serological and virological studies were undertaken to investigate these cases with the aim of determining the genotype of the mumps virus (MuV) circulating in Turkey. Samples from 23 cases-Ankara (n:5), Kirklareli (n:4), Mugla (n:10), Isparta (n:3), Trabzon (n:1)-with a diagnosis of clinical parotitis were investigated. Serum samples were tested against mumps IgM and IgG, nested PCR amplification of a 639-bp fragment encompassing the entire SH gene was performed using buccal swabs, and PCR products were sequenced. Of 18 serum samples, 16 (88.9%) were positive for mumps IgM. Seven (30.4%) of 23 buccal swab samples were positive by PCR. In five PCR-positive cases, the sample was also positive for mumps IgM, and serum samples were not available from two of the PCR-positive cases. There was 98% identity between the different sequences, and all were identified as genotype H. The sequences were most similar to sequences identified in Spain, Japan, Switzerland and the UK, and less closely related to the H strains identified in Belarus, Korea and Russia. This is the first report of the mumps virus genotypes circulating in Turkey. Turkey is, geographically, a bridge between Europe and Asia, and therefore, a better understanding of the molecular epidemiology of MuV in Turkey may led to improved tracking of the circulation of strains between the two continents. Moreover, there is a need to further investigate the existence of other genotypes in Turkey.

A prolonged mumps outbreak among highly vaccinated Aboriginal people in the Kimberley region of Western Australia

To describe a prolonged outbreak of mumps in the Kimberley region of Western Australia in 2007-2008. Descriptive analysis of all mumps cases notified to the WA Notifiable Infectious Diseases Database for the period 1 July 2007 to 30 June 2008. Notified cases of mumps by patients' place of residence, age, Indigenous or non-Indigenous ethnicity, vaccination status and method of diagnosis. 84% (153/183) of mumps notifications in WA over the study period occurred in the Kimberley region or were directly linked to Kimberley cases. Median age of patients was 18 years (range, 2-63 years), and 54% of patients were aged less than 20 years. Almost all (92%) were Australian Aboriginal people; 67% (102/153) had received at least one dose of mumps vaccine, and 52% had received two doses. The highest notification rate (1816 cases per 100,000 population) was in the Aboriginal 15-19-years age group, and 92% of these patients had received at least one dose of mumps vaccine. Almost all outbreak cases (94%) were laboratory confirmed. Genotyping was performed on 20 mumps virus isolates: all were genotype J. A prolonged outbreak of mumps occurred in a well defined, highly vaccinated, predominantly young Aboriginal population in the remote Kimberley region of WA. This outbreak raises questions about the effectiveness and scheduling of the current vaccine (which is genotype A-derived), especially for Aboriginal people. Surveillance of circulating mumps virus genotypes and neutralisation studies will help in evaluating the protection provided by the current vaccine against genotypically different strains.

Molecular identification of two genotypes of mumps virus causing two regional outbreaks in Asturias, Spain

Background In spite of universal vaccination, several sporadic cases of mumps infection, which could produce outbreaks, are detected every year in different countries. Objective Mumps virus strains causing two regional outbreaks in Asturias (Spain) were phylogenetically characterized. Study design Mumps virus strains, which were detected in samples from patients belonging to two regional outbreaks in Asturias, were characterized by sequencing of the SH gene and further alignment to homologous sequences of representative strains of the different mumps genotypes. Results Two different strains (Ast/SP02 and Ast/SP07) were isolated. Sequence analysis revealed that while Ast/SP02 belonged to genotype H, Ast/SP07 was phylogenetically close to UK02-19, a reference strain for a new genotype. Both strains belonged to different genotypes from those used in the vaccination (Jeryl-Lynn strain is genotype A). Conclusion Mumps virus strains different from those used in vaccination program can cause mumps outbreaks even in vaccinated patients.

Variability of hemagglutinin-neuraminidase and nucleocapsid protein of vaccine and wild-type mumps virus strains

The mumps virus (MuV) molecular evolution is characterized by the co-circulation of numerous distinct strains. Standardized phylogenetic analyses based on the nucleotide sequences of the SH gene are important for mumps surveillance, but lack the information regarding antigenic properties. So far, the location of antigenic epitopes has been determined for two MuV proteins, the hemagglutinin-neuraminidase (HN) and the nucleocapsid (N) protein. We performed multiple sequence comparisons of putative HN and N protein sequences in order to describe their diversity and plasticity, and to determine the level of similarity between vaccine and wild-type strains. The results of full-length HN or N protein phylogeny showed that MuV strains form a number of differing clades which are in concordance with grouping obtained by standard MuV genotyping. When vaccine strains are compared to all wild-type strains, the highest mean percentage of amino acid differences in both HN and N protein analysis was found for Jeryl Lynn 5 and Jeryl Lynn 2 strains while the lowest value was obtained for Leningrad-3 and L-Zagreb strains. When only 3 antigenic regions of the HN protein, comprising 45 amino acids in total, were investigated, the diversity is considerably diminished: 51.5% of all putative HN proteins show identical sequences (including those of vaccine strains L-Zagreb, Leningrad-3, Hoshino and Urabe). Another 26.5% proteins (including Miyahara vaccine strain) differ in only one amino acid, while the others differ in two to five amino acids from the most common sequence. Jeryl Lynn 2 and Jeryl Lynn 5 strains differ in four amino acids each. N protein antigenic sites have been mapped within its hypervariable C-terminus. Our results indicate that there might be genotype-specific amino acids residing in this antigenic region. The results of our study present the background information for investigations of MuV heterogeneity and antigenic diversity.

Detection of a new mumps virus genotype during parotitis epidemic of 2006–2007 in the State of São Paulo, Brazil

Nucleotide sequence analyses of the SH gene of 18 mumps virus isolates collected in the 2006-2007 parotitis epidemic in the state of São Paulo identified a new genotype, designated genotype M. This new designation fulfills all the parameters required to define a new mumps virus genotype. The parameters were established by an expert panel in collaboration with the World Health Organization (WHO) in 2005. This information will enhance the mumps virus surveillance program both at the national and global levels.