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Chinese Journal of Pediatrics ; (12): 30-35, 2022.
Article in Chinese | WPRIM | ID: wpr-935635


Objective: To compare the clinical characteristics of different types of human adenovirus (HAdV) infection in hospitalized children with acute respiratory infection in Beijing, and to clarify the clinical necessity of adenovirus typing. Methods: In a cross-sectional study, 9 022 respiratory tract specimens collected from hospitalized children with acute respiratory infection from November 2017 to October 2019 in Affiliated Children's Hospital, Capital Institute of Pediatrics were screened for HAdV by direct immunofluorescence (DFA) and (or) nucleic acid detection. Then the Penton base, Hexon and Fiber gene of HAdV were amplified from HAdV positive specimens to confirm their HAdV types by phylogenetic tree construction. Clinical data such as laboratory results and imaging data were analyzed for children with predominate type HAdV infection using t, U, or χ2 test. Results: There were 392 cases (4.34%) positive for HAdV among 9 022 specimens from hospitalized children with acute respiratory infection. Among those 205 cases who were successfully typed, 131 were male and 74 were female, age of 22.6 (6.7, 52.5) months,102 cases (49.76%) were positive for HAdV-3 and 86 cases (41.95%), HAdV-7, respectively, while 17 cases were confirmed as HAdV-1, 2, 4, 6, 14 or 21. In comparison of clinical characteristics between the predominate HAdV type 7 and 3 infection, significant differences were shown in proportions of children with wheezing (10 cases (11.63%) vs. 25 cases (24.51%)), white blood cell count >15 ×109/L (4 cases (4.65%) vs.14 cases (13.73%)), white blood cell count <5×109/L (26 cases (30.23%) vs.11 cases (10.78%)), procalcitonin level>0.5 mg/L (43 cases (50.00%) vs. 29 cases (28.43%)), multilobar infiltration (45 cases (52.33%) vs.38 cases (37.25%)), pleural effusion (23 cases (26.74%) vs. 10 cases (9.80%)), and severe adenovirus pneumonia (7 cases (8.14%) vs. 2 cases (1.96%)) with χ²=5.11, 4.44, 11.16, 9.19, 4.30, 9.25, 3.91 and P=0.024, 0.035, 0.001, 0.002, 0.038, 0.002, 0.048, respectively, and also in length of hospital stay (11 (8, 15) vs. 7 (5, 13) d, Z=3.73, P<0.001). Conclusions: HAdV-3 and 7 were the predominate types of HAdV infection in hospitalized children with acute respiratory tract infection in Beijing. Compared with HAdV-3 infection, HAdV-7 infection caused more obvious inflammatory reaction, more severe pulmonary symptoms, longer length of hospital stay, suggesting the clinical necessity of further typing of HAdVs.

Adenovirus Infections, Human/epidemiology , Adenoviruses, Human/genetics , Beijing/epidemiology , Child , Child, Hospitalized , Cross-Sectional Studies , Female , Humans , Infant , Male , Phylogeny , Respiratory Tract Infections/epidemiology
Article in Chinese | WPRIM | ID: wpr-935240


Objective: To express DNA-binding protein (DBP) of human adenovirus (HAdV) type 7 using the prokaryotic expression system, and product anti-HAdV-7 DBP rabbit polyclonal antibody. Methods: The HAdV-7 DBP gene was synthesized and cloned into prokaryotic expressing vector pET30a, and the recombinant plasmid was transformed into E. coli BL21 (DE3) competent cell. The recombinant protein DBP was expressed by induced Isopropyl-beta-D-thiogalactopyranoside (IPTG) and purified with Ni-NTA affinity column. The titer of anti-DBP polyclonal antibody produced in immunized rabbit was measured by indirect ELISA, and the specificity of the antibody was identified by Western blotting and indirect immunofluorescence assay (IFA). In addition, purified rDBP was used as coating antigen for indirect ELISA assay to detect specific IgM and IgG antibodies against DBP in the serum of children infected with HAdV. Results: The HAdV-7 DBP plasmid was constructed successfully. The purified recombinant DBP was more than 95% after purification. The titer of polyclonal antibody was 1∶1 024 000. The polyclonal antibody showed high specificity in vitro using Western blotting and IFA. The positive rate of specific anti-DBP IgM and IgG antibody in acute-phase serum samples collected from children infected with HAdV were 50.0% (19/38) and 63.2% (24/38), respectively, using indirect ELISA. Conclusion: In summary, the HAdV-7 rDBP is expressed using prokaryotic expression system, and the recombinant HAdV-7 DBP protein and the anti-DBP rabbit polyclonal antibody with high titer are prepared.

Adenoviruses, Human/genetics , Animals , Antibody Specificity , Blotting, Western , DNA-Binding Proteins/metabolism , Enzyme-Linked Immunosorbent Assay , Escherichia coli/genetics , Immunoglobulin G , Rabbits
Chinese Journal of Biotechnology ; (12): 1824-1836, 2022.
Article in Chinese | WPRIM | ID: wpr-927820


In order to construct a recombinant replication deficient human type 5 adenovirus (Ad5) expressing a foot-and-mouth disease virus (FMDV) capsid protein, specific primers for P12A and 3B3C genes of FMDV-OZK93 were synthesized. The P12A and 3B3C genes were then amplified and connected by fusion PCR, and a recombinant shuttle plasmid pDC316-mCMV-EGFP-P12A3B3C expressing the FMDV-OZK93 capsid protein precursor P12A and 3B3C protease were obtained by inserting the P12A3B3C gene into the pDC316-mCMV-EGFP plasmid. The recombinant adenovirus rAdv-P12A3B3C-OZK93 was subsequently packaged, characterized and amplified using AdMaxTM adenovirus packaging system, and the expression was verified by infecting human embryonic kidney cell HEK-293. The humoral and cellular immunity levels of well-expressed and purified recombinant adenovirus immunized mice were evaluated. The results showed that rAdv-P12A3B3C-OZK93 could be stably passaged and the maximum virus titer reached 1×109.1 TCID50/mL. Western blotting and indirect immunofluorescence showed that rAdv-P12A3B3C-OZK93 expressed the FMDV-specific proteins P12A and VP1 in HEK-293 cells. In addition, the PK cell infection experiment confirmed that rAdv-P12A3B3C-OZK93 could infect porcine cells, which is essential for vaccination in pigs. Comparing with the inactivated vaccine group, the recombinant adenovirus could induce higher FMDV-specific IgG antibodies, γ-IFN and IL-10. This indicates that the recombinant adenovirus has good immunity for animal, which is very important for the subsequent development of foot-and-mouth disease vaccine.

Adenoviridae/genetics , Adenoviruses, Human/genetics , Animals , Antibodies, Viral , Capsid/metabolism , Capsid Proteins , Foot-and-Mouth Disease/prevention & control , Foot-and-Mouth Disease Virus/genetics , HEK293 Cells , Humans , Mice , Recombinant Proteins/genetics , Serogroup , Swine , Viral Proteins , Viral Vaccines/genetics
Rev. Soc. Bras. Med. Trop ; 50(5): 621-628, Sept.-Oct. 2017. tab, graf
Article in English | LILACS | ID: biblio-897017


Abstract INTRODUCTION: Acute gastroenteritis (AGE) is one of the most common causes of morbidity and mortality, especially among children from developing countries. Human adenovirus (HAdV) and sapovirus (SaV) are among the agents that cause AGE. The present study aimed to detect and genotype HAdV and SaV in 172 fecal samples from children with AGE, collected during a surveillance study carried out in a low-income community in Belém, Pará, between 1990 and 1992. METHODS: HAdV was detected by nested PCR, using primers Hex1deg/Hex2deg and NeHex3deg/NeHex4deg. SaV was assayed by reverse transcription PCR (RT-PCR), nested PCR, and quantitative PCR. The nucleotide sequence was determined by direct cycle sequencing. RESULTS: Overall, 43% (74/172) of samples were positive for HAdV, of which 70.3% (52/74) were sequenced and classified as belonging to five different species, mostly A and F. For SaV, positivity was 5.2% (9/172) and genotypes GI.1, GI.7, GII.1, and GV.2 were detected. CONCLUSIONS: The present results reinforce the need for further studies to obtain epidemiological data about the circulation of these viruses in Brazil, especially in the Amazon Region, where data from the early 1990's are scarce. Furthermore, the study describes for the first time the detection of SaV genotypes GI.7 and GV.2 in Brazil, showing that these types circulated in the region more than 25 years ago.

Humans , Male , Female , Infant, Newborn , Infant , Child, Preschool , Brazil/epidemiology , Adenoviruses, Human/isolation & purification , Caliciviridae Infections/virology , Sapovirus/isolation & purification , Gastroenteritis/virology , Genotype , Phylogeny , Time Factors , Base Sequence , Adenovirus Infections, Human/epidemiology , Adenovirus Infections, Human/virology , Adenoviruses, Human/genetics , Polymerase Chain Reaction , Prevalence , Prospective Studies , Age Distribution , Caliciviridae Infections/epidemiology , Sapovirus/genetics , Genotyping Techniques/methods , Gastroenteritis/enzymology , Genes, Viral
Braz. j. microbiol ; 47(1): 243-250, Jan.-Mar. 2016. tab, graf
Article in English | LILACS | ID: lil-775111


Abstract Human adenovirus species F (HAdV-F) type 40 and 41 are commonly associated with acute diarrheal disease (ADD) across the world. Despite being the largest state in southeastern Brazil and having the second largest number of inhabitants, there is no information in the State of Minas Gerais regarding the role of HAdV-F in the etiology of ADD. This study was performed to determine the prevalence, to verify the epidemiological aspects of infection, and to characterize the strains of human adenoviruses (HAdV) detected. A total of 377 diarrheal fecal samples were obtained between January 2007 and August 2011 from inpatient and outpatient children of age ranging from 0 to 12 years. All samples were previously tested for rotavirus, norovirus, and astrovirus, and 314 of 377 were negative. The viral DNA was extracted, amplified using the polymerase chain reaction and the HAdV-positive samples were sequenced and phylogenetically analyzed. Statistical analyses were performed using the Chi-square test (p < 0.05), considering two conditions: the total of samples tested (377) and the total of negative samples for the remaining viruses tested (314). The overall prevalence of HAdV was 12.47% (47/377); and in 76.60% (36/47) of the positive samples, this virus was the only infectious agent detected. The phylogenetic analysis of partial sequences of 32 positive samples revealed that they all clustered with the HAdV-F type 41. The statistical analysis showed that there was no correlation between the onset of the HAdV infection and the origin of the samples (inpatients or outpatients) in the two conditions tested: the total of samples tested (p = 0.598) and the total of negative samples for the remaining viruses tested (p = 0.614). There was a significant association in the occurrence of infection in children aged 0–12 months for the condition 1 (p = 0.030) as well as condition 2 (p = 0.019). The occurrence of infections due to HAdV did not coincide with a pattern of seasonal distribution. These data indicate the significant involvement of HAdV-F type 41 in the etiology of ADD in Minas Gerais, which demonstrates the importance of other viral agents in the development of the disease after the introduction of rotavirus vaccine immunization.

Child , Child, Preschool , Humans , Infant , Infant, Newborn , Adenovirus Vaccines/administration & dosage , Adenoviruses, Human/isolation & purification , Diarrhea/epidemiology , Diarrhea/prevention & control , Rotavirus Infections/epidemiology , Rotavirus Infections/prevention & control , Adenovirus Vaccines/immunology , Adenoviruses, Human/classification , Adenoviruses, Human/genetics , Brazil/epidemiology , Cluster Analysis , DNA, Viral/chemistry , DNA, Viral/genetics , DNA, Viral/isolation & purification , Feces/virology , Genotype , Phylogeny , Prevalence , Sequence Analysis, DNA
Braz. j. microbiol ; 46(3): 749-752, July-Sept. 2015.
Article in English | LILACS | ID: lil-755826


Human adenoviruses (HAdV), members of the Adenoviridae family, are excreted through the fecal route and may be present in the feces of humans consuming contaminated food or water. The presence of HAdV from different serotypes in the feces of healthy individuals was already reported using conventional polymerase chain reaction; however, real-time PCR (qPCR) may reveal not only the rates of detection as well as demonstrate the viral loads excreted by healthy persons. Aiming to identify and characterize the presence of adenoviruses in stool samples, 147 fecal samples from patients with no records of diarrhea were analyzed (74 from winter season and 73 from summer) by Real-Time PCR (qPCR) assay and conventional PCR. HAdV genome was present in 43.8% (32/73) of stools samples collected during summer season and 21.6% (16/74) during winter. The rate of detection of genomic copies (gc) ranged from 4.04×102 to 6.72×105gc/g of feces among the 147 samples analyzed, of which the ranged of genomic copies of DNA HAdV was major in summer. All samples were negative when tested for rotaviruses (RV) and noroviruses (NoV) by PCR conventional and qPCR respectively. HAdV is excreted constantly by infected individuals in the absence of clinical signs and the occurrence may vary seasonally.


Adult , Female , Humans , Male , Middle Aged , Young Adult , Adenovirus Infections, Human/epidemiology , Adenoviruses, Human/genetics , Adenoviruses, Human/isolation & purification , Feces/virology , Adenovirus Infections, Human/virology , Adenoviruses, Human/classification , Brazil , Norovirus/genetics , Norovirus/isolation & purification , Real-Time Polymerase Chain Reaction , Rotavirus/genetics , Rotavirus/isolation & purification , Seasons
Mem. Inst. Oswaldo Cruz ; 109(4): 428-435, 03/07/2014. tab, graf
Article in English | LILACS | ID: lil-716303


Viral acute gastroenteritis (AG) is a significant cause of hospitalisation in children younger than five years. Group A rotavirus (RVA) is responsible for 30% of these cases. Following the introduction of RVA immunisation in Brazil in 2006, a decreased circulation of this virus has been observed. However, AG remains an important cause of hospitalisation of paediatric patients and only limited data are available regarding the role of other enteric viruses in these cases. We conducted a prospective study of paediatric patients hospitalised for AG. Stool samples were collected to investigate human adenovirus (HAdV), RVA, norovirus (NoV) and astrovirus (AstV). NoV typing was performed by nucleotide sequencing and phylogenetic analysis. From the 225 samples tested, 60 (26%) were positive for at least one viral agent. HAdV, NoV, RVA and AstV were detected in 16%, 8%, 6% and 0% of the samples, respectively. Mixed infections were found in nine patients: HAdV/RVA (5), HAdV/NoV (3) and HAdV/NoV/RVA (1). The frequency of fever and lymphocytosis was significantly higher in virus-infected patients. Phylogenetic analysis of NoV indicated that all of these viruses belonged to genotype GII.4. The significant frequency of these pathogens in patients with AG highlights the need to routinely implement laboratory investigations.

Child , Humans , DNA Virus Infections/virology , Feces/virology , Gastroenteritis/virology , Acute Disease , Adenoviruses, Human/genetics , Adenoviruses, Human/isolation & purification , Brazil , Genotype , Hospitalization , Mamastrovirus/genetics , Mamastrovirus/isolation & purification , Norovirus/genetics , Norovirus/isolation & purification , Phylogeny , Prospective Studies , Real-Time Polymerase Chain Reaction , RNA, Viral/genetics , Rotavirus/genetics , Rotavirus/isolation & purification , Seasons
Article in English | IMSEAR | ID: sea-144678


Background & objectives: Though several viruses are responsible for conjunctivitis, but human adenovirus (HAdV) is by far the most common cause. Epidemic conjunctivitis causes morbidity and early detection of aetiological agent is essential in preventing spread of disease as some of serotypes of adenoviruses cause a severe form of conjunctivitis. This study was undertaken to identify the causative agent of conjunctivitis outbreak in Chennai in 2010. Methods: Conjunctival samples collected from 17 patients with conjunctivitis were subjected to virological investigations. Culture and PCR for detection of adenovirus and enterovirus were carried out. PCR positive products were further subjected for DNA sequencing. The nucleotide sequences of the hexons of isolates were analyzed by comparison with all 51 human adenovirus strains. Phylogenetic tree was constructed using DAMBE software. Results: Among 17 patients, seven were positive for adenovirus by PCR on the direct specimen, none was positive for enterovirus. Eleven of 30 conjunctival swabs showed cytopathic effect in HEp-2 cell line and were confirmed as HAdV by PCR. The DNA sequence data of the 11 isolates had equal percentage of homology with HAdV 6 and 2 on blast analysis. On phylogenetic analysis with GeneBank data of 51 adenovirus strains, 11 isolates from patients during the outbreak of conjunctivitis formed a separate clade indicating a new variant strain. Interpretation & conclusions: Based on phylogenetic analysis it was concluded that the recent conjunctivitis outbreak that occurred in Chennai was caused by a variant adenovirus strain.

Adenoviruses, Human/genetics , Adenoviruses, Human/isolation & purification , Disease Outbreaks , Humans , India/epidemiology , Keratoconjunctivitis/diagnosis , Keratoconjunctivitis/epidemiology , Polymerase Chain Reaction/methods , Phylogeography
Rev. Inst. Med. Trop. Säo Paulo ; 52(6): 317-321, Nov.-Dec. 2010. graf, tab
Article in English | LILACS | ID: lil-570730


Adenovirus (AdV) respiratory infections are usually described as being associated with high mortality rates. Laboratory diagnosis is essential for the establishment of the appropriate therapy, and for guiding the implementation of preventive measures in order to prevent the spread of the infection. Aiming to analyze the sensitivity and specificity of the laboratorial diagnosis methods available, we compared antigen detection by indirect immunofluorescence assay (IF), and a specific nested polymerase chain reaction (PCR), to detect AdV in respiratory samples collected from patients admitted to hospital with acute respiratory disease. Positive samples were inoculated into a cell culture to confirm the results. We analyzed 381 samples from the nasopharyngeal aspirates collected during the year 2008; of these, 2.6 percent tested were positive for adenovirus through IF and 10 percent through PCR; positive isolation was obtained in 40 percent and 26 percent of these cases, respectively. Most infected patients were children under six months of age, and despite of the fact that a significant number of patients required intensive care, the mortality rate was low (5 percent). In conclusion, molecular methods were found to be useful for rapid diagnosis of adenovirus infections with higher sensitivity than antigen detection; their introduction permitted a significant increase in diagnoses of adenovirus infections.

Infecções respiratórias por Adenovírus (ADV) são geralmente descritas associadas com alta mortalidade. O diagnóstico laboratorial é essencial para o estabelecimento da terapêutica adequada e para orientar a implantação de medidas preventivas evitando a propagação da infecção. Com o objetivo de analisar a sensibilidade e a especificidade dos métodos de avaliação de diagnóstico laboratorial, foi comparada a detecção de antígeno por imunofluorescência indireta (IF) com a reação em cadeia da polimerase específica (PCR) para detectar AdV em amostras respiratórias coletadas de pacientes internados com doença respiratória aguda. As amostras com resultados positivos foram inoculadas em cultura celular. Foram analisadas 381 amostras da secreção nasofaríngea coletadas durante o ano de 2008, das quais 2,6 por cento foram positivas pela IF e 10 por cento pela PCR, isolamento positivo foi obtido em 40 por cento e 26 por cento dos casos positivos pelos testes anteriores, respectivamente. A maioria dos pacientes infectados eram crianças com menos de seis meses de idade, e apesar do fato de que um número significativo de pacientes necessitou de cuidados intensivos, a taxa de mortalidade foi baixa (5 por cento). Em conclusão, os métodos moleculares são úteis para o diagnóstico rápido de infecções por adenovírus com maior sensibilidade do que a detecção do antígeno, a sua introdução na rotina permitiu um aumento significativo no diagnóstico de infecções por adenovírus.

Child , Child, Preschool , Humans , Infant , Adenoviruses, Human , Adenovirus Infections, Human/diagnosis , Nasopharynx/virology , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/virology , Adenoviruses, Human/genetics , Adenoviruses, Human/immunology , Adenoviruses, Human/isolation & purification , Cross-Sectional Studies , Fluorescent Antibody Technique, Indirect , Polymerase Chain Reaction , Prospective Studies , Sensitivity and Specificity
Rev. chil. infectol ; 27(2): 148-152, abr. 2010. ilus, tab
Article in Spanish | LILACS | ID: lil-548130


Introduction: Eye infection is a common cause of ophtalmologic consultation. Adenovirus keratoconjunctivitis outbreaks are common worldwide but its impact and clinical characteristic in Chilean population is unkown. Objective: To describe a series of adenovirus keratoconjunctivitis cases. Patients and Method: The Índex case and contacts received medical care in the Hospital Clínico Universidad de Chile between April and August 2006. A complete ophthalmologic exam and microbiologic evaluation was performed. Results: Nine patients presented a pattern of characteristic epidemic keratoconjunctivitis. In x cases sub-corneal epithelial infiltrates were observed for a period of more than six months. Three affected patients were ophtalmologists, staff at the Hospital. In seven patients ADV was isolated all bellonging to type D genus. Conclusions: Adenovirus type D caused epidemic keratoconjunctivitis in a series of Chilean individuals. Ophthalmologist may have transmitted the virus to patients.

Introducción: La patología ocular infecciosa es frecuente en la consulta oftalmológica, especialmente la conjuntivitis y queratoconjuntivitis epidémica (QCE). Brotes de esta patología son causados por adenovirus (ADV) en el extranjero; en Chile se desconoce su impacto y características. Objetivos: Describir una serie de casos de queratonconjuntivitis epidémica por adenovirus. Material y Pacientes: Al caso índice y los contactos de una serie de casos de QCE por ADV que consultaron en el Hospital Clínico de la Universidad de Chile, entre abril y agosto de 2006, se les realizó examen oftalmológico completo y estudio de ADV por aislamiento viral, detección de antígenos y de genoma viral. Se estableció el género de ADV mediante reacción de polimerasa en cadena. Resultados: Los 9 pacientes infectados presentaron QCE característica. En algunos casos se observaron infiltrados sub-epiteliales corneales que se extendieron por más de seis meses. Tres pacientes eran médicos oftalmólogos. En 7 de los 9 pacientes examinados se aisló ADV; todos del género D. Conclusiones: En Chile, la QCE puede ser causada por el subgénero tipo D. El médico oftalmólogo es un potencial vector en la transmisión de ADV en un brote de QCE, por lo que es fundamental que sea considerado en las estrategias de prevención de esta patología.

Female , Humans , Male , Adenovirus Infections, Human/transmission , Adenoviruses, Human/isolation & purification , Cross Infection/virology , Disease Outbreaks , Keratoconjunctivitis/virology , Acute Disease , Adenovirus Infections, Human/diagnosis , Adenovirus Infections, Human/epidemiology , Adenoviruses, Human/genetics , Cross Infection/diagnosis , Cross Infection/epidemiology , Disease Outbreaks/statistics & numerical data , Infectious Disease Transmission, Professional-to-Patient , Keratoconjunctivitis/diagnosis , Keratoconjunctivitis/epidemiology
Rev. Inst. Med. Trop. Säo Paulo ; 51(4): 227-229, July-Aug. 2009. ilus, graf
Article in English | LILACS | ID: lil-524380


Viral conjunctivitis is a common, highly contagious disease often caused by adenovirus. We investigate the frequency of adenoviral conjunctivitis in the population of Rio de Janeiro, Brazil, between March 2004 and May 2007 and identified the predominant serotype circulating among this population. Seventy-five ocular swabs were collected from 66 patients with clinical presentation of conjunctivitis. The specimens were analyzed for detection of adenovirus (AdV) by polymerase chain reaction (PCR). The PCR products were further analyzed for virus typing by sequence analysis and/or heteroduplex mobility assay (HMA). Forty-five samples (60%) were positive for AdV of which 21 samples were typed as AdV19 (46.7%), 7 AdV8 (15.5%), 3 AdV31 (6.7%), and one each AdV1, AdV2, AdV3, AdV4 and AdV6. For nine samples the serotype was not determined. AdV19 was the predominant serotype circulating in Rio de Janeiro during the studied period.

A conjuntivite viral é doença ocular comum, altamente contagiosa, frequentemente causada por adenovírus. Neste estudo, investigamos a frequência de conjuntivite por adenovírus na população do Rio de Janeiro, Brasil, entre março de 2004 e maio de 2007, e identificamos o sorotipo predominante circulando nesta população. Setenta e cinco swabs de secreção ocular foram coletados de 66 pacientes com conjuntivite. As amostras foram analisadas para detecção de adenovírus (AdV) por reação em cadeia da polimerase (PCR). Os produtos da PCR foram caracterizados por sequenciamento e/ou ensaio de mobilidade do heteroduplex (Heteroduplex Mobility Assay - HMA) para identificação do sorotipo viral. Quarenta e cinco (60%) amostras foram positivas para AdV das quais 21 foram identificadas como pertencentes ao sorotipo AdV19 (46,7%), sete AdV8 (15,5%), três AdV31 (6,7%), e uma de cada: AdV1, AdV2, AdV3, AdV4 e AdV6. Para nove amostras o sorotipo não pode ser identificado. O AdV 19 foi o sorotipo predominante circulando no Rio de Janeiro durante o período estudado.

Humans , Adenovirus Infections, Human/epidemiology , Adenoviruses, Human/genetics , Conjunctivitis, Viral/epidemiology , DNA, Viral/genetics , Adenoviruses, Human/classification , Brazil/epidemiology , Conjunctiva/virology , Conjunctivitis, Viral/virology , Molecular Epidemiology , Polymerase Chain Reaction , Serotyping
Braz. j. microbiol ; 40(1): 102-107, Jan.-Mar. 2009. graf, tab
Article in English | LILACS | ID: lil-513124


The aim of this study was to verify the presence and annual distribution of adenoviruses and hepatitis A virus in domestic sewage in the city of Limeira, São Paulo. Fifty samples with a volume of 8 liters each were collected weekly from December 2004 to December 2005. The viruses were concentrated by filtration through positively charged ZP60S filter membranes, followed by ultracentrifugation. Human adenoviruses (HAdV) were detected by PCR followed by nested-PCR and screening for species F was done by restriction of the PCR product with TaqI endonuclease. Virus infectivity assays were performed by inoculation of concentrates onto HEp-2 cell monolayers. RT-PCR was used for the detection of hepatitis A virus. HAdV were detected in all samples, and 64% of samples were positive for infectious virus. Species F was present in 82% of the samples. Hepatitis A virus was detected in 48% of the samples. These results demonstrate that HAdV and HAV were present in the domestic sewage of Limeira throughout the period of study, demonstrating the importance of an adequate treatment before the disposal in the environment.

O objetivo do estudo foi verificar a ocorrência e a distribuição anual de adenovírus humanos e vírus da Hepatite A (VHA) no efluente doméstico da cidade de Limeira, São Paulo, ao longo do período de Dezembro de 2004 e Dezembro de 2005, com vistas à futura implementação de sistemas de tratmento de água de esgoto. Cinquenta amostras de efluente bruto com volume de 8L cada foram colhidas semanalmente e os vírus concentrados por filtração em membrana eletropositiva ZP60S, seguida de ultracentrifugação. Adenovírus foram detectados por PCR e nested-PCR. Adenovírus da espécie F foram distinguidos das demais por restrição do produto da PCR com endonuclease TaqI. Ensaios de infectividade viral foram realizados em culturas de células HEp-2. A presença do vírus da hepatite A também foi pesquisada nas mesmas amostras, fazendo-se uso de método de RT-PCR. Adenovírus foram detectados em todas as amostras, sendo a espécie F identificada em 82% destas. Sessenta e quatro por cento dos adenovírus detectados ainda estavam infecciosos. O vírus da Hepatite A foi detectado em 48% das amostras examinadas. Estes resultados evidenciam a presença e a circulação de Adenovírus humano e VHA nas águas de esgoto doméstico de Limeira ao longo do período de estudo, demonstrando a importância de um tratamento adequado desse material antes da disposição no meio ambiente.

Humans , Adenoviruses, Human/genetics , Adenoviruses, Human/isolation & purification , Wastewater/analysis , Endonucleases/analysis , Membrane Filtration/analysis , In Vitro Techniques , Polymerase Chain Reaction , Water Purification/analysis , Hepatitis A Virus, Human/genetics , Hepatitis A Virus, Human/isolation & purification , Measures of Disease Occurrence , Methods , Methods , Water Samples
Mem. Inst. Oswaldo Cruz ; 103(2): 195-200, Mar. 2008. graf, tab
Article in English | LILACS | ID: lil-480634


Adenoviruses (AdV) are commonly involved in acute respiratory infections (ARI), which cause high morbidity and mortality in children. AdV are grouped in six species (A-F), which are associated with a wide range of diseases. The aim of this study was to identify the AdV species infecting non-hospitalized Mexican children with ARI symptoms, attending to the same school. For that, a PCR/RFLP assay was designed for a region of the hexon gene, which was chosen, based on the bioinformatical analysis of AdV genomes obtained from GenBank. A total of 100 children's nasopharyngeal samples were collected from January to June, 2005, and used for viral isolation in A549 cells and PCR/RFLP analysis. Only 15 samples produced cytopathic effect, and in all of them AdV C was identified. AdV C was also identified in eight additional nasopharyngeal samples which were negative for viral isolation. In summary, this outpatient population showed a rate of AdV infection of 23 percent, and only AdV C was detected.

Child , Female , Humans , Male , Adenovirus Infections, Human/virology , Adenoviruses, Human/genetics , Respiratory Tract Infections/virology , Acute Disease , Adenovirus Infections, Human/diagnosis , Adenovirus Infections, Human/epidemiology , Adenoviruses, Human/isolation & purification , Bacterial Typing Techniques , DNA Restriction Enzymes/analysis , Genetic Markers , Genome, Viral , Mexico/epidemiology , Nasopharynx/virology , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Respiratory Tract Infections/diagnosis , Respiratory Tract Infections/epidemiology , Seasons
Mem. Inst. Oswaldo Cruz ; 102(8): 937-941, Dec. 2007. ilus
Article in English | LILACS | ID: lil-471860


From January to December 1998, nasopharyngeal aspirates were obtained from 482 children with acute respiratory infections attended in emergence department and wards of a teaching hospital in the city of Salvador, Brazil. The samples were tested for the presence of adenovirus by isolation in tissue culture and indirect immunofluorescence assay. Eleven adenoviruses were detected by both methods in the same clinical samples. Infections by adenovirus were observed during seven months of the year without association with rainy season. Genome analysis was performed on these 11 isolates. Species C was represented by serotypes 1, 2 and 5. Within species B, only serotype 7 (Ad7) was detected. Two genomic variants of Ad1, two variants of Ad2, one of Ad5, and one of Ad7 (7h) were identified. This is the first study of molecular epidemiology of adenovirus associated to acute respiratory infections in children living in Northeast Brazil, and contributes to a better understanding of adenovirus infections in the country.

Child , Humans , Adenoviruses, Human , Adenovirus Infections, Human/virology , Respiratory Tract Infections/virology , Acute Disease , Adenovirus Infections, Human/epidemiology , Adenoviruses, Human/genetics , Adenoviruses, Human/immunology , Adenoviruses, Human/isolation & purification , Brazil/epidemiology , DNA, Viral/analysis , Fluorescent Antibody Technique, Indirect , Genotype , Nasopharynx/virology , Respiratory Tract Infections/epidemiology
Medicina (B.Aires) ; 65(3): 196-200, 2005. ilus, tab
Article in Spanish | LILACS | ID: lil-425260


El objetivo de este trabajo fue determinar la prevalencia de adenovirus (ADV) en las infecciones del sistema nervioso central (SNC). Se analizaron 108 muestras de líquido cefalorraquídeo (LCR) provenientes de 79 casos de encefalitis, 7 meningitis y 22 de otras patologías neurológicas, recibidas en el período 2000-2002. Cuarenta y nueve (47.35%) se obtuvieron de pacientes inmunocomprometidos. La presencia de ADV se investigó mediante reacción en cadena de la polimerasa en formato anidado (Nested-PCR). La identificación del genogrupo se realizó mediante análisis filogenético de la secuencia nucleotídica parcial de la región que codifica para la proteína del hexón. Se detectó la presencia de ADV en 6 de 108 (5.5%) muestras de LCR analizadas. Todos los casos positivos pertenecieron a pacientes con encefalitis que fueron 79, (6/79, 7.6%). No se observó diferencia estadísticamente significativa entre los casos de infección por ADV en pacientes inmunocomprometidos e inmunocompetentes (p>0.05). Las cepas de ADV detectadas se agruparon en los genogrupos B1 y C. En conclusión, nuestros resultados describen el rol de los ADV en las infecciones neurológicas en Argentina. La información presentada contribuye al conocimiento de su epidemiología, en particular en casos de encefalitis.

Infant, Newborn , Infant , Child, Preschool , Child , Adolescent , Adult , Middle Aged , Aged, 80 and over , Humans , Male , Female , Adenovirus Infections, Human/virology , Adenoviruses, Human/isolation & purification , Central Nervous System Infections/virology , Adenovirus Infections, Human/classification , Adenovirus Infections, Human/genetics , Adenoviruses, Human/genetics , Central Nervous System Infections/classification , Central Nervous System Infections/genetics , Encephalitis, Viral/virology , Polymerase Chain Reaction , Prospective Studies
Indian J Pathol Microbiol ; 2004 Apr; 47(2): 302-5
Article in English | IMSEAR | ID: sea-74607


Polymerase chain reaction (PCR) was evaluated to detect Adenoviruses and Chlamydia trachomatis on nasopharyngeal aspirates (NPA) obtained 4-5 days after the onset of lower respiratory tract illness in children. Forty-five nasopharyngeal aspirates (NPA) from 45 children with lower respiratory tract infections were processed for the detection of C. trachomatis and Adenovirus by Fluorescent antibody test (FAT), culture and PCR for the cryptic plasmid of C. trachomatis and the gene coding for hexon of Adenoviruses. Seven (13.3%) and 4 (6.6%) of the 45 specimens were positive for C. trachomatis and adenovirus by PCR respectively, which included one specimen each positive for these agents. Cultures were negative for both the organisms. PCRs showed a statistically significant (McNemar test--p= 0.004) higher sensitivity. PCR test is necessary to detect C. trachomatis and adenovirus in nasopharyngeal aspirates obtained 4-5 days after the onset of illness.

Adenovirus Infections, Human/diagnosis , Adenoviruses, Human/genetics , Base Sequence , Child, Preschool , Chlamydia Infections/diagnosis , Chlamydia trachomatis/genetics , DNA, Bacterial/genetics , DNA, Viral/genetics , Humans , Infant , Nasopharynx/microbiology , Polymerase Chain Reaction/methods , Respiratory Tract Infections/microbiology , Sensitivity and Specificity , Suction
Mem. Inst. Oswaldo Cruz ; 93(2): 231-5, Mar.-Apr. 1998. ilus, tab, graf
Article in English | LILACS | ID: lil-203604


During an epidemiological survey of acute respiratory infection in Rio de Janeiro, among 208 adenovirus isolates, we found two strains that we were not able, by a standard neutralization procedure, to distinguish between type 3 or 7. However, DNA restriction pattern for the two strains with different enzymes were analysed and showed a typical Ad3h profile. Using a cross-neutralization test in which both Ad3p and Ad7p antisera were used in different concentration against 100 TCID 50 of each adenovirus standard and both isolates, we were able to confirm that the two isolates belong to serotype 3. An hemagglutination inhibition test also corroborated the identification of both strains as adenovirus type 3. Comparing Ad3h and Ad3p genome, we observed 16 different restriction enzyme sites, three of which were located in genomic regions encoding polypeptides involved in neutralization sites.

Humans , Adenoviruses, Human/genetics , Neutralization Tests