RESUMO
Human adenovirus (HAdV) is a significant viral pathogen causing severe acute respiratory infections (SARIs) in children. To improve the understanding of type distribution and viral genetic characterization of HAdV in severe cases, this study enrolled 3404 pediatric SARI cases from eight provinces of China spanning 2017-2021, resulting in the acquisition of 112 HAdV strains. HAdV-type identification, based on three target genes (penton base, hexon, and fiber), confirmed the diversity of HAdV types in SARI cases. Twelve types were identified, including species B (HAdV-3, 7, 55), species C (HAdV-1, 2, 6, 89, 108, P89H5F5, Px1/Ps3H1F1, Px1/Ps3H5F5), and E (HAdV-4). Among these, HAdV-3 exhibited the highest detection rate (44.6%), followed by HAdV-7 (19.6%), HAdV-1 (12.5%), and HAdV-108 (9.8%). All HAdV-3, 7, 55, 4 in this study belonged to dominant lineages circulating worldwide, and the sequences of the three genes demonstrated significant conservation and stability. Concerning HAdV-C, excluding the novel type Px1/Ps3H1F1 found in this study, the other seven types were detected both in China and abroad, with HAdV-1 and HAdV-108 considered the two main types of HAdV-C prevalent in China. Two recombinant strains, including P89H5F5 and Px1/Ps3H1F1, could cause SARI as a single pathogen, warranting close monitoring and investigation for potential public health implications. In conclusion, 5 years of SARI surveillance in China provided crucial insights into HAdV-associated respiratory infections among hospitalized pediatric patients.
Assuntos
Infecções por Adenovirus Humanos , Adenovírus Humanos , Infecções Respiratórias , Criança , Humanos , Adenovírus Humanos/genética , Análise de Sequência de DNA/métodos , Filogenia , Adenoviridae/genética , China/epidemiologia , Infecções Respiratórias/epidemiologiaRESUMO
Human adenovirus (HAdV) is the primary cause of acute conjunctivitis. To improve our understanding of the etiology of adenoviral conjunctivitis, ocular samples were collected from 160 conjunctivitis cases in the Shanxi province of northern China between 2016 and 2019. Through preliminary identification, virus isolation, and type identification, a total of 63 HAdV isolates were obtained from the samples. Three species and seven types (HAdV-3, HAdV-4, HAdV-8, HAdV-37, HAdV-53, HAdV-64, and HAdV-85) were detected, with HAdV-64, HAdV-3, and HAdV-8 being the predominant types in 2016, 2018, and 2019, respectively. Further phylogenetic analysis indicated the relative genomic stability of seven HAdV-type strains, except for 4 HAdV-3 strains in 2018 with a novel amino acid insertion site (Pro) between P19 and S20 in the penton base gene. It is worth noting that the genomes of two Shanxi HAdV-85 strains from 2016 were almost identical to those of previously reported HAdV-85 strains that circulated in Japan in 2014 to 2018. China was the second country to sample and isolate HAdV-85, suggesting that HAdV-85 might be underreported as an ocular pathogen. Data obtained in this study provide valuable information on the prevalence of acute conjunctivitis caused by HAdV. IMPORTANCE HAdV types in cases of conjunctivitis in Shanxi province, China, in 2016 to 2019 showed evident diversity, with seven types (HAdV-3, HAdV-4, HAdV-8, HAdV-37, HAdV-53, HAdV-64, and HAdV-85) being identified, and relative genome stability of these viruses was observed. In addition, China was the second country to sample and isolate HAdV-85, which suggests that HAdV-85 might be underreported as an important pathogen associated with ocular infections. These results enhance the understanding of the etiology of adenoviral conjunctivitis and may aid in the development of prevention and control strategies for HAdV-related ocular infections in China.
Assuntos
Infecções por Adenovirus Humanos , Adenovírus Humanos , Conjuntivite , Infecções Oculares , Humanos , Filogenia , China/epidemiologia , Infecções por Adenovirus Humanos/epidemiologia , Doença AgudaRESUMO
To understand the molecular evolution of human parainfluenza virus type 2 (HPIV2), 21 Hemagglutinin-Neuraminidase (HN) gene sequences covering seven Chinese provinces in 2011 and 2017 to 2021 were combined with 90 published HN sequences worldwide for phylogenetic analysis. The result showed that global HPIV2 could be classified into two distinct clusters (I and II), five lineages (IA to IIE), and four sublineages (IB1 and 2, and IIE1 and 2). The minimum genetic distances between different clusters and lineages were 0.049 and 0.014, respectively. In the last decade, one lineage (IID) and three sublineages (IB1, IB2, and IIE1) have been cocirculating in China, with the sublineages IB2 and IIE1 dominating, while sublineages IB1 and IIE1 are dominant globally. In addition, the spread of HPIV2 had relative spatial clustering, and sublineage IB2 has only been detected in China thus far. The overall evolution rate of HPIV2 was relatively low, on the order of 10-4 substitutions/site/year, except for sublineage IB2 at 10-3 substitutions/site/year. Furthermore, human-animal transmission was observed, suggesting that the HPIV2 might have jumped out of animal reservoirs in approximately 1922, the predicted time of a common ancestor. The entire HN protein was under purifying/negative selection, and the specific amino acid changes and two novel N-glycosylation sites (N316 and N517) in sublineages IB1, IB2, and IIE1 were mostly located in the globular head region of the HN protein. In this study, preliminary evolutionary characteristics of HPIV2 based on the HN gene were obtained, increasing the recognition of the evolution and adaptation of HPIV2. IMPORTANCE The phylogenetic analysis showed that global HPIV2 could be classified into two distinct clusters (I and II) and five lineages (IA to IIE) with at least 0.049 and 0.014 genetic distances between clusters and lineages, respectively. Furthermore, lineages IB and IIE could be further divided into two sublineages (IB1-2 and IIE1-2). All China sequences belong to one lineage and three sublineages (IB1, IB2, IID, and IIE1), among which sublineages IB2 and IIE1 are predominant and cocirculating in China, while sublineages IB1 and IIE1 are dominant globally. The overall evolution rate of HPIV2 is on the order of 10-4 substitutions/site/year, with the highest rate of 2.18 × 10-3 for sublineage IB2. The entire HN protein is under purifying/negative selection, and the specific amino acid substitutions and two novel N-glycosylation sites (N316 and N517) in sublineages IB1, IB2, and IIE1 are mostly located in the globular head region of the HN protein.
Assuntos
Vírus da Parainfluenza 2 Humana , Neoplasias do Colo do Útero , Animais , Feminino , Humanos , Vírus da Parainfluenza 2 Humana/genética , Vírus da Parainfluenza 2 Humana/metabolismo , Filogenia , Neuraminidase , Hemaglutininas/metabolismo , Proteína HN/genética , Proteína HN/metabolismo , Proteínas Virais/genética , Evolução MolecularRESUMO
BACKGROUND: Outbreaks of severe, acute hepatitis among children have recently attracted global attention. The pathogen causing the outbreak remains unknown, but there is growing evidence that it may be associated with human adenovirus (HAdV). DATA SOURCES: A review of adenovirus-related clinical studies, epidemiological studies, etiological studies, and case reports was conducted by reviewers independently. RESULTS: HAdV can cause a wide variety of clinical symptoms. In the Mainland of China, HAdV infection accounts for 5.8%-13% of patients with acute respiratory infections, and these infections are mainly caused by species B, C, and E of HAdV. For acute conjunctivitis, 39.8%-74.9% of sporadic cases were infected by B and D species of HAdV. Outbreaks of keratoconjunctivitis and pharyngoconjunctival fever related to HAdV infection could be found throughout the country. In pediatric patients with acute gastroenteritis, HAdV-41 was the predominant HAdV type, followed by HAdV species B and C. Several types of HAdV, including HAdV-5, HAdV-7, HAdV-1, and HAdV-2, have previously been reported as potential pathogens associated with HAdV hepatitis in immunocompromised patients. However, few HAdV-related hepatitis cases have been reported in China to date. CONCLUSIONS: There are no systematic surveillance and clinical studies on HAdV hepatitis in China. Therefore, it is imperative to establish a nationwide HAdV virological surveillance system to collect relevant clinical, epidemiological and virological surveillance data and risk factor information as soon as possible to assess the potential risk of HAdV hepatitis among children.
Assuntos
Infecções por Adenoviridae , Infecções por Adenovirus Humanos , Infecções Respiratórias , Doença Aguda , Infecções por Adenoviridae/epidemiologia , Infecções por Adenovirus Humanos/diagnóstico , Infecções por Adenovirus Humanos/epidemiologia , Criança , China/epidemiologia , Humanos , Filogenia , Infecções Respiratórias/diagnósticoRESUMO
[This corrects the article DOI: 10.1371/journal.pone.0232092.].
RESUMO
Human adenovirus (HAdV-7) is a highly contagious pathogen that causes severe respiratory illnesses. However, the epidemic patterns and genetic variability of HAdV-7 circulating in mainland China have not been well elucidated. In this study, we used Chinese HAdV sentinel surveillance data obtained from 2012-2015 to investigate the clinical features of 122 HAdV-7-positive cases and performed amplification and sequence determination of three capsid genes (penton base, hexon, and fiber) from 69 isolated viruses covering from seven provinces of China. Additionally, we compared with data from representative sequences of 21 strains covering seven more provinces in China and 32 international HAdV-7 strains obtained from GenBank database to determine the phylogenetic, sequence variations, and molecular evolution of HAdV-7. The results indicated that HAdV-7 infection occurred throughout the year, and a high proportion of severe cases (27 cases, 22.1%) exhibited infantile pneumonia. Moreover, phylogenetic analysis showed that all HAdV-7 strains could be divided into two major evolutionary branches, including subtype 1 and subtype 2, and subtype 3 was also formed according to analysis of the penton base gene. Subtypes 1 and 2 co-circulated in China before 2008, and HAdV-7 strains currently circulating in China belonged to subtype 2, which was also the predominant strain circulating worldwide in recent years. Further sequence variation analysis indicated that three genes of HAdV-7 were relatively stable across time and geographic space, particularly for viruses within subtypes, which shared almost the same variation sites. Owing to continuous outbreaks caused by HAdV-7, resulting in increased illness severity and fatality rates in China, the establishment of a national HAdV surveillance system is urgently needed for the development of effective preventive and infection-control interventions for adenovirus respiratory infections in China.
Assuntos
Infecções por Adenovirus Humanos/genética , Adenovírus Humanos/genética , Proteínas do Capsídeo/genética , Adenoviridae/genética , Infecções por Adenovirus Humanos/epidemiologia , Infecções por Adenovirus Humanos/virologia , China/epidemiologia , Surtos de Doenças , Evolução Molecular , Variação Genética/genética , Humanos , Filogenia , Infecções Respiratórias/epidemiologia , Análise de Sequência de DNA/métodosRESUMO
The human mastadenovirus C (HAdV-C) cause respiratory infections in children. Homologous recombination was clearly involved in the molecular evolution of HAdV-A, B, and D, but little is known about the molecular evolution of HAdV-C. From 2000 to 2016, 201 HAdV-C strains were collected from nine provinces covering six administrative regions of mainland of China via 3 existing surveillance programs, namely the febrile respiratory syndrome surveillance, the acute flaccid paralysis surveillance, and the hand, foot, and mouth disease surveillance system. The genes coding for the capsid protein (penton base, hexon, and fiber) of 201 HAdV-C strains were sequenced and compared with representative sequences publicly available. In addition, the whole genome sequence of 24 representative strains of HAdV-C was generated for further recombination analysis. Phylogenetic analysis of the penton base sequences of HAdV-C revealed six genetic groups (labelled as Px1-6), which showed that the penton base had more variation than previously thought. Based on the penton base, hexon, and fiber gene sequences, 16 new genetic patterns of HAdV-C circulating in mainland of China were identified in this study. Whole genome sequence analysis revealed frequent recombination events among HAdV-C genomes. This study is highly beneficial for case classification, tracking the transmission chain, and further epidemiological exploration of HAdV-C-related severe clinical diseases in the near future. Our data demonstrated that multiple newly divergent HAdV-C co-circulated across mainland China during the research period.
Assuntos
Infecções por Adenovirus Humanos/diagnóstico , Adenovírus Humanos/classificação , Proteínas do Capsídeo/genética , Sequenciamento Completo do Genoma/métodos , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/genética , Adenovírus Humanos/isolamento & purificação , Linhagem Celular , Pré-Escolar , China , Evolução Molecular , Tamanho do Genoma , Doença de Mão, Pé e Boca/virologia , Humanos , Lactente , Paraplegia/virologia , Filogenia , Vigilância da População , Infecções Respiratórias/virologia , Análise de Sequência de DNA/métodosRESUMO
To date, at least three lineages (Lineage 1-3) that are related to recombinant human adenovirus species C (HAdV-C) have been identified in China. Among them, Lineage 1 includes two Chinese strains, strain KR699642-CHN-20093 (CBJ11) and strain MF315029-CHN-2013 (BJ09), which were collected in Beijing in 2009 and 2013, respectively. Herein, we performed genomic and bioinformatics analysis of two HAdV-C strains (strain SX-2000-140 and strain SX-2004-327) that were isolated from the feces of two healthy children in Shanxi province of China in 2000 and 2004, respectively. Results revealed that the genomes of both Shanxi strains had the highest homology to two Chinese HAdV-C strains belonging to Lineage 1 and harbored the genetic elements of these two strains, thereby presuming that Lineage1 has been circulated in mainland of China for decades. In addition, though the viruses in Lineage 1 showed slightly different recombinant patterns resulting from the recombinant events among the five types of HAdV-C, all the Lineage 1 viruses shared the highest sequence similarities with the HAdV-2 prototype strain (NC_001405-USA-1953) across the genome, especially in the major capsid genes including hexon, and fiber. These results indicated that Lineage 1 viruses that were associated with recombinants shared a common ancestor that is closely related to the HAdV-2 virus. Our current findings confirmed that frequent recombination among the different HAdV-C types might be an important driving force for the molecular evolution of HAdV-C. Therefore, there is a strong need for further comprehensive and systematic monitoring, detection, and research on HAdV-C.
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Infecções por Adenovirus Humanos/epidemiologia , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/genética , Recombinação Genética , Adenovírus Humanos/classificação , China/epidemiologia , Biologia Computacional/métodos , Evolução Molecular , Genoma Viral , Genômica/métodos , Humanos , Anotação de Sequência Molecular , Filogenia , Análise de Sequência de DNARESUMO
This study aims at analyzing all publicly available HAdV-C whole genome sequences (WGSs) and describes the genetic relationships between these genomes as well as identifies potential hotspots for recombination throughout the viral genome. In addition to the 4 prototypical genomic sequences, this analysis identified 20 HAdV-C WGSs which should be relevant for future recombination analysis of HAdV-C. This report confirmed the recombinogenic property of HAdV-C genomes and identified two main regions for breakpoints, within the hexon gene and around the fiber genomic region. No obvious recombination was detected between HAdV-Cs and non-human mastadenoviruses or non-C HAdVs. Finally, it highlighted the need for a surveillance of HAdVs in order to detect novel recombinant types that might represent health risks and develop possible prevention measures. Genetic analyses of recombination between recently collected HAdV-Cs and the assessment of their potential virulence are necessary steps towards the establishment of a surveillance of HAdVs in the future.
Assuntos
Adenovírus Humanos/genética , Genoma Viral , Recombinação Genética , Infecções por Adenovirus Humanos/virologia , Adenovírus Humanos/classificação , Adenovírus Humanos/patogenicidade , DNA Viral/genética , Evolução Molecular , Humanos , Filogenia , Análise de Sequência de DNA , Virulência/genética , Sequenciamento Completo do GenomaRESUMO
Human mastadenovirus species C (HAdV-C) are the most common etiologic agents of respiratory disease in young children and are frequently detected worldwide including China. Two recombinant HAdV-C strains (BJ04 and BJ09) were isolated from infants with acute respiratory infection (ARI) in Beijing in 2012-2013. The whole genome sequences (WGS) of BJ04 and BJ09 were generated and compared to other 35 HAdV-C WGSs publicly available. Phylogenetic analyses showed that the BJ04 strain might be the result of three homologous recombination events involving the parental strains JX173086 (HAdV-1), NC_001405 (HAdV-2) and LC068718 (HAdV-6), whereas BJ09 viral genome might be made of genetic elements from JX173083 (HAdV-1), KF268199 (HAdV-5), and KR699642 (strain CBJ113). Despite intratypic recombination, amino acid analysis showed that the gene repertoire of BJ04 and BJ09 were similar to type 2 viruses. Finally, this analysis revealed that at least three lineages of HAdV-C have been identified in China, represented by BJ04 related to NC_001405, BJ09 related to CBJ113, and KF951595 (strain DD28) related to virus isolated in Japan. This study showed that the frequent recombination played an important driving force for complexity of the HAdV-C epidemic in Beijing, thereby demonstrating the necessity for epidemiological and virological surveillance for HAdV-C in China.
Assuntos
Infecções por Adenovirus Humanos/genética , Genoma Viral , Recombinação Genética , Infecções Respiratórias/genética , Infecções Respiratórias/virologia , Infecções por Adenovirus Humanos/epidemiologia , Adenovírus Humanos , Pequim/epidemiologia , Epidemias , Feminino , Estudo de Associação Genômica Ampla , Humanos , Lactente , Japão/epidemiologia , Masculino , Infecções Respiratórias/epidemiologiaRESUMO
Virologic surveillance is a critical component of measles management. One of the criteria for verification of elimination of endemic measles is genetic analysis of wild-type viruses to demonstrate lack of an indigenous genotype. Measles is yet to be eliminated in China, and genotype H1 has been detected continuously since virologic surveillance was initiated in 1993. Virologic surveillance has been very active in China, providing a unique opportunity to conduct a detailed study of the evolution of a single, endemic genotype over a timespan of nearly two decades. Phylogenetic analysis performed on the 450 nt coding sequence for the C-terminal 150 amino acids of the nucleoprotein (N-450), fusion (F) gene and haemagglutinin (H) gene confirmed the continued circulation of genotype H1 viruses for 19 years. No evidence of selective pressure for the H protein was found. The substitution rates ranged from 0.75×10(-3) substitutions site(-1) year(-1) for H to 1.65×10(-3) substitutions site(-1) year(-1) for N-450. The time of most recent common ancestor (TMRCA) for genotype H1 was estimated as approximately 1985 (95â% highest probability density, 1979-1989). Finally, the overall diversity of measles sequences from China decreased from 2005 to 2012, coincident with a substantial decrease in measles cases. The results suggest that detailed evolutionary analyses should facilitate the documentation of eventual measles elimination in China. Moreover, the molecular approaches used in this study can be applied in other countries approaching measles elimination.
Assuntos
Hemaglutininas Virais/genética , Vírus do Sarampo/genética , Sarampo/epidemiologia , Proteínas Virais de Fusão/genética , Proteínas Virais/genética , Animais , Sequência de Bases , Evolução Biológica , Callithrix , Linhagem Celular , China/epidemiologia , Chlorocebus aethiops , Variação Genética , Genótipo , Sarampo/virologia , Vírus do Sarampo/classificação , Filogenia , Alinhamento de Sequência , Análise de Sequência de RNA , Células VeroRESUMO
To characterize the genetic properties of coxsackievirus A12 (CVA12) strains isolated from hand, foot and mouth disease (HFMD) patients in Qingdao during 2008-2011, the complete genome and VP1 coding region were sequenced and analyzed. Phylogenetic analysis showed that all strains from China clustered into three different branches, suggesting multiple lineages of CVA12 co-circulating in Asia. Sequence analysis indicated a monophyletic group only when the P1 region was examined, indicating possible recombination between CVA12 and other HEV-A serotypes. The emergence of CVA12 involved in an HFMD outbreak in China is a public-health issue.
Assuntos
Doenças Transmissíveis Emergentes/virologia , Enterovirus/classificação , Enterovirus/genética , Doença de Mão, Pé e Boca/virologia , RNA Viral/genética , Pré-Escolar , China/epidemiologia , Análise por Conglomerados , Doenças Transmissíveis Emergentes/epidemiologia , Enterovirus/isolamento & purificação , Feminino , Genótipo , Doença de Mão, Pé e Boca/epidemiologia , Humanos , Lactente , Masculino , Epidemiologia Molecular , Dados de Sequência Molecular , Filogenia , Recombinação Genética , Análise de Sequência de DNA , Proteínas Estruturais Virais/genéticaRESUMO
BACKGROUND: Acute respiratory infections (ARIs) are the leading cause of children and their leading killer. ARIs are responsible for at least six percent of the world's disability and death. Viruses are one of the most common agents causing ARIs. Few studies on the viral etiology and clinical characteristics of ARIs have been performed in the northwest region of China, including Gansu Province. METHODS: Clinical and demographic information and throat swabs were collected from 279 patients from January 1st to December 30st, 2011. Multiplex RT-PCR was performed to detect 16 respiratory viral pathogens. RESULTS: 279 patients were admitted for ARIs. The patients aged from 1 month to 12 years, with the median age of 2 years. Of which, 105 (37.6%) were positive for at least one pathogen. A total of 136 respiratory viral pathogens were identified from the 105 patients. Respiratory syncytial virus (RSV) was the most frequently detected pathogen (26.5%, 36/136), followed by parainfluenza virus (PIV) 1-3 (22.1%, 30/136), human rhinovirus (HRV) (21.3%, 29/136), human coronavirus (CoV) (10.3%, 14/136) and human adenovirus (HAdV) (9.6%, 13/136). Influenza A (Flu A), human metapneumovirus (hMPV) and human bocavirus (BoCA) were found 4.4%, 3.7% and 2.2%, respectively. Influenza B (Flu B) and seasonal influenza A H1N1(sH1N1) were not detected. Single-infections were detected in 30.5% (85/279) of cases. RSV was the most common pathogens in patients under 1 year and showed seasonal variation with peaks during winter and spring. CONCLUSIONS: This paper presents data on the epidemiology of viral pathogens associated with ARIs among children in Gansu Province, China. RSV is most frequently detected in our study. The findings could serve as a reference for local CDC in drawing up further plans to prevent and control ARIs.
Assuntos
Infecções Respiratórias/epidemiologia , Infecções Respiratórias/virologia , Doença Aguda/epidemiologia , Distribuição por Idade , Criança , Pré-Escolar , China/epidemiologia , Feminino , Humanos , Lactente , Recém-Nascido , Masculino , Infecções Respiratórias/etiologia , Infecções Respiratórias/prevenção & controle , Estações do AnoRESUMO
Large-scale Hand, Foot, and Mouth Disease (HFMD) outbreaks have frequently occurred in China since 2008, affecting more than one million children and causing several hundred children deaths every year. The pathogens of HFMD are mainly human enteroviruses (HEVs). Among them, human enterovirus 71 (HEV71) and coxsackievirus A16 (CVA16) are the most common pathogens of HFMD. However, other HEVs could also cause HFMD. To rapidly detect HEV71 and CVA16, and ensure detection of all HEVs causing HFMD, two real-time hybridization probe-based RT-PCR assays were developed in this study. One is a multiplex real-time RT-PCR assay, which was developed to detect and differentiate HEV71 specifically from CVA16 directly from clinical specimens within 1-2 h, and the other is a broad-spectrum real-time RT-PCR assay, which targeted almost all HEVs. The experiments confirmed that the two assays have high sensitivity and specificity, and the sensitivity was up to 0.1 TCID50/ml for detection of HEVs, HEV71, and CVA16, respectively. A total of 213 clinical specimens were simultaneously detected by three kinds of assays, including the two real-time RT-PCR assays, direct conventional RT-PCR assay, and virus isolation assay on human rhabdomyosarcoma cells (RD cells). The total positive rate of both HEV71 and CVA16 was 69.48% with real-time RT-PCR assay, 47.42% with RT-PCR assay, and 34.58% with virus isolation assay. One HFMD clinical specimen was positive for HEV, but negative for HEV71 or CVA16, which was identified as Echovirus 11 (Echo11) by virus isolation, RT-PCR, and sequencing for the VP1 gene. The two real-time RT-PCR assays had been applied in 31 provincial HFMD labs to detect the pathogens of HFMD, which has contributed to the rapid identification of the pathogens in the early stages of HFMD outbreaks, and helped to clarify the etiologic agents of HFMD in China.
Assuntos
Doença de Mão, Pé e Boca/virologia , Reação em Cadeia da Polimerase em Tempo Real , Criança , China/epidemiologia , Surtos de Doenças , Enterovirus/isolamento & purificação , Enterovirus Humano A/isolamento & purificação , Doença de Mão, Pé e Boca/epidemiologia , Doença de Mão, Pé e Boca/mortalidade , Humanos , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e EspecificidadeRESUMO
A GeXP based multiplex RT-PCR assay was developed to simultaneously detect twelve different respiratory viruses types/subtypes including influenza A virus, influenza B virus, influenza A virus sH1N1, parainfluenza virus type 1, parainfluenza virus type 2, parainfluenza virus type 3, human rhinovirus, human metapneumovirus, adenovirus, respiratory syncytial virus A, respiratory syncytial virus B and human bocavirus. Twelve sets of specific primers were designed based on the conserved sequences of available respiratory-virus sequence database. The specificity of the multiplex system was examined by positive specimens confirmed previously. The sensitivity to detect twelve respiratory viruses simultaneously was 10(3) copies/microL. Twenty four clinical specimens were further detected by this novel assay and the results were compared with that of the real-time RT-PCR. These results showed that this novel assay based on GeXP is a fast, sensitive, and high throughput test for the detection of respiratory virus infections.
Assuntos
Vírus de RNA/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Infecções Respiratórias/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Humanos , Vírus da Influenza A Subtipo H1N1/genética , Vírus da Influenza A Subtipo H1N1/isolamento & purificação , Orthomyxoviridae/genética , Orthomyxoviridae/isolamento & purificação , Infecções por Orthomyxoviridae/virologia , Vírus de RNA/genética , Reação em Cadeia da Polimerase em Tempo Real/instrumentação , Vírus Sinciciais Respiratórios/genética , Vírus Sinciciais Respiratórios/isolamento & purificação , Reação em Cadeia da Polimerase Via Transcriptase Reversa/instrumentação , Rhinovirus/genética , Rhinovirus/isolamento & purificação , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: An outbreak of aseptic meningitis occurred in Tianshui city of Gansu Province, the People's Republic of China, from March to June 2005. A total of 85 patients were clinical confirmed as aseptic meningitis in this outbreak. RESULTS: CVA9 was mainly responsible for this outbreak supported by the clinical manifestations of the patients, epidemiological data of the outbreak, the results of RT-PCR and complete VP1 sequence determination, conventional neutralization assays, IgM serological assays, viral isolation and phylogenetics analysis. Through phylogenetic analysis and homogeneity analysis for partial VP1 gene, the nucleotide and amino acid homologies between Gansu isolates and former Chinese CVA9 strains were 88.2%-96.1% and 97.2%-99.2%, respectively. Multiple transmission chains of CVA9 occurred in different provinces or years in China. Moreover, in order to clarify the genotype of CVA9, Gansu CVA9 strains isolated in this outbreak were compared with other CVA9 isolates based on VP1/2A junction regions (genotyping region) and they might belong to a new genotype of CVA9, which could be assigned for genotype XIII, CONCLUSIONS: CVA9 was confirmed as the pathogen responsible for this outbreak. The phylogenetic analysis indicated that the CVA9 strains isolated in this outbreak might belong to a new genotype.
Assuntos
Infecções por Coxsackievirus/epidemiologia , Surtos de Doenças , Enterovirus Humano B/isolamento & purificação , Meningite Asséptica/epidemiologia , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Criança , Pré-Escolar , China/epidemiologia , Análise por Conglomerados , Infecções por Coxsackievirus/virologia , Enterovirus Humano B/classificação , Enterovirus Humano B/genética , Enterovirus Humano B/imunologia , Humanos , Imunoglobulina M/sangue , Meningite Asséptica/virologia , Dados de Sequência Molecular , Filogenia , RNA Viral/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Análise de Sequência de DNA , Homologia de Sequência de Aminoácidos , Homologia de Sequência do Ácido Nucleico , Proteínas Estruturais Virais/genéticaRESUMO
An outbreak of aseptic meningitis (AM) occurred in Jinzhai County in Anhui province from April to July in 2005. Totally, 97 children aged 3-15 years were hospitalized. To identify the etiologic agent, 77 cerebrospinal fluid specimens (CSF) and 5 fecal specimens were collected from the patients and cultured by human rhabdomyosarcoma (RD) cell line. Thirty isolates of human echovirus 6 (E6) from 27 CSF and 3 fecal specimens were confirmed by neutralization assay and sequencing analysis of the VP1 gene. The homology of VP1 gene among Anhui isolates was 99.7-100.0% and it indicated that this AM outbreak probable caused by a single transmission link of E6. Phylogenetic analysis based on all the available complete VP1 sequences indicated that E6 could be divided into clusters A, B, and C with at least 15% diversity between clusters and the C cluster could be further divided into C1, C2, C3, and C4. The Anhui isolates most resembled a 2005 strain from Russia (25465 Tambov) and belong to C4. This is the first report that E6 was responsible for an outbreak of AM in China. J. Med. Virol. 82:441-445, 2010. (c) 2010 Wiley-Liss, Inc.
Assuntos
Surtos de Doenças , Echovirus 6 Humano/isolamento & purificação , Infecções por Echovirus/epidemiologia , Meningite Asséptica/epidemiologia , Adolescente , Proteínas do Capsídeo/genética , Técnicas de Cultura de Células , Linhagem Celular , Líquido Cefalorraquidiano/virologia , Criança , Pré-Escolar , China/epidemiologia , Análise por Conglomerados , Fezes/virologia , Hospitalização , Humanos , Dados de Sequência Molecular , Testes de Neutralização , Filogenia , Análise de Sequência de DNA , Homologia de Sequência de AminoácidosRESUMO
An outbreak of acute respiratory tract infection occurred in Shanxi Province, China, from March to April 2006. Of the 254 patients affected by this outbreak, 247 patients were students of a senior high school; 1 of these patients died during the outbreak. Serological tests and blood culture revealed no evidence of bacterial infection. The results of direct reverse transcription-PCR or PCR performed with clinical specimens collected from the patients, including the sole patient who died, were positive for human adenoviruses (HAdVs) but negative for influenza virus, measles virus, rubella virus, mumps virus, parainfluenza virus, respiratory syncytial virus, and human enteroviruses. These findings were confirmed by enzyme-linked immunosorbent assay for HAdV immunoglobulin A, the conventional neutralization test, and viral isolation and identification. Sequencing of the entire hexon gene revealed that HdAV type 11a (HAdV-11a) belonging to the B2 species of HAdV was the etiological agent responsible for the outbreak. However, both the analysis of the phylogenetic relationship and the similarity plot indicated that the sequence of the 3' end of the hexon gene outside the hypervariable regions the HAdV-11a strain isolated in this outbreak may be a recombinant with the sequence of the HAdV-14 strain of species B2. Although isolates of HAdV species B2 seldom cause respiratory infections, they may pose a new global challenge with regard to acute respiratory diseases; this possibility cannot be overlooked and should be carefully considered. Hence, the need to establish and improve both epidemiological and virological surveillance of HAdV infections in China should be emphasized.
Assuntos
Infecções por Adenoviridae/diagnóstico , Infecções por Adenoviridae/epidemiologia , Adenoviridae/classificação , Adenoviridae/isolamento & purificação , Surtos de Doenças , Infecções Respiratórias/epidemiologia , Adenoviridae/genética , Infecções por Adenoviridae/mortalidade , Adolescente , Anticorpos Antivirais/análise , China/epidemiologia , DNA Viral/química , DNA Viral/genética , Ensaio de Imunoadsorção Enzimática , Humanos , Imunoglobulina A/análise , Dados de Sequência Molecular , Testes de Neutralização , Reação em Cadeia da Polimerase/métodos , Infecções Respiratórias/mortalidade , Infecções Respiratórias/virologia , Análise de Sequência de DNA , EstudantesRESUMO
OBJECTIVE: To understand the prevalence of Epstein-Barr virus (EBV) infection in urban and rural areas of Beijing using the serological method. METHODS: Totally 589 serum samples were collected from children in Beijing urban and rural areas who were 0--14 years old and tested with Viron-Seron ELISA classic EBV virus capsid antigen IgG antibody (EBV VCA IgG) kit. Optical density of serum samples was obtained at the wavelength of 405 nanometers. Sero-positive or negative samples were determined according to standard curve and cut-off attached in ELISA classic EBV VCA IgG kits. The activity of EBV VCA IgG was calculated by using special formula. The percentage and activity of EBV VCA IgG from Beijing children were compared with SPSS 13.0 between the urban and rural areas. RESULTS: The percentage of EBV VCA IgG seropositive samples was 83.6%, and 80.8% in those from urban and 86.2% in those from rural areas. The peak value of EBV infection was 71% seen among children under the age of 3 years, and in urban area the rate was 67.7%, which was lower than that in the rural area (75.3%), and was 82.5% by the age of 6, which was lower than the data (up to 90%) reported 30 years ago. There was a significant difference in EBV infection rate and VCA IgG activities in children at different ages between urban and rural areas (P < 0.05). CONCLUSION: The rate of EBV infection in children living in urban area was lower by the age of 6 years. The primary infection of EBV occurred late in part of children lived in urban area.