RESUMO
Sapovirus (SV) is one of the major causative agents of viral gastroenteritis affecting all age groups worldwide. A new method for the quantitative detection of SV from clinical stool specimens by real-time reverse transcription-polymerase chain reaction (RT-PCR) based on TaqMan MGB technology was described. Primers and probe were designed to target the RNA-dependent RNA polymerase/capsid genes junction. Performance of the newly developed assay was validated against a panel of 244 clinical stool specimens collected for patients with gastroenteritis. SV was detected in eight (3.3%) specimens. Phylogenetic analysis of the positive isolates suggested that the assay could detect at least SV genogroups I, II and IV. In addition, the assay had an increased detection rate compared with a widely used conventional RT-PCR assay. Quantitative analysis showed that the assay could detect as low as 10 copies of viral cDNA per reaction. No cross-reactivity with norovirus and rotavirus was observed. In conclusion, the assay is a sensitive and specific method for the detection of SV from clinical stool specimens.
Assuntos
Infecções por Caliciviridae/virologia , Fezes/virologia , Gastroenterite/virologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Sapovirus/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Bases , Capsídeo , Criança , Pré-Escolar , Primers do DNA , Feminino , Genes Virais/genética , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , RNA Viral/genética , RNA Polimerase Dependente de RNA/genética , Sapovirus/classificação , Sapovirus/genética , Sensibilidade e Especificidade , Alinhamento de Sequência , Especificidade da Espécie , Ensaio de Placa ViralRESUMO
Pulmonary infection caused by the opportunistic organisms Penicillium marneffei and Stenotrophomonas maltophilia in patients with Job's syndrome is rare and not well documented. The case of a 30-year-old man with Job's syndrome who developed recurrent pneumonia and lung abscesses caused by P. marneffei and S. maltophilia, complicated by massive hemoptysis, is described. Bronchial artery embolization was successful in controlling the hemoptysis; however, the infection proved fatal despite appropriate antimicrobial therapy. A brief review of the literature on Job's syndrome and its associated infective pulmonary manifestations is also presented.
Assuntos
Infecções por Bactérias Gram-Negativas/complicações , Hemoptise/microbiologia , Síndrome de Job/microbiologia , Pneumopatias Fúngicas/complicações , Penicillium/isolamento & purificação , Stenotrophomonas maltophilia/isolamento & purificação , Adulto , Humanos , Síndrome de Job/complicações , MasculinoRESUMO
We report the median cDNA viral load of norovirus genogroup II is >100-fold higher than that of genogroup I in the fecal specimens of patients with norovirus-associated gastroenteritis. We speculate that increased cDNA viral load accounts for the higher transmissibility of genogroup II strains through the fecal-oral route.
Assuntos
Fezes/virologia , Gastroenterite/virologia , Norovirus/isolamento & purificação , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Infecções por Caliciviridae/virologia , Criança , Pré-Escolar , DNA Complementar/análise , DNA Viral/análise , Feminino , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Norovirus/classificação , Norovirus/genética , Carga ViralRESUMO
We evaluated an indirect immunofluorescence assay based on virus-infected cells for detecting anti-severe acute respiratory syndrome-associated coronavirus (SARS-CoV) immunoglobulin (Ig) G antibody. All confirmed SARS cases demonstrated seroconversion or fourfold rise in IgG antibody titer; no control was positive. Sensitivity and specificity of this assay were both 100%. Immunofluorescence assay can ascertain the status of SARS-CoV infection.
Assuntos
Doenças Transmissíveis Emergentes/diagnóstico , Técnica Indireta de Fluorescência para Anticorpo/métodos , Síndrome Respiratória Aguda Grave/diagnóstico , Adolescente , Adulto , Anticorpos Antivirais/isolamento & purificação , Criança , Pré-Escolar , Doenças Transmissíveis Emergentes/imunologia , Doenças Transmissíveis Emergentes/fisiopatologia , Feminino , Hong Kong , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Síndrome Respiratória Aguda Grave/imunologia , Síndrome Respiratória Aguda Grave/fisiopatologiaRESUMO
The virologic test results of 415 patients with severe acute respiratory syndrome (SARS) were examined. The peak detection rate for SARS-associated coronavirus occurred at week 2 after illness onset for respiratory specimens, at weeks 2 to 3 for stool or rectal swab specimens, and at week 4 for urine specimens. The latest stool sample that was positive by reverse transcription-polymerase chain reaction (RT-PCR) was collected on day 75 while the patient was receiving intensive care. Tracheal aspirate and stool samples had a higher diagnostic yield (RT-PCR average positive rate for first 2 weeks: 66.7% and 56.5%, respectively). Pooled throat and nasal swabs, rectal swab, nasal swab, throat swab, and nasopharyngeal aspirate specimens provided a moderate yield (29.7%-40.0%), whereas throat washing and urine specimens showed a lower yield (17.3% and 4.5%). The collection procedures for stool and pooled nasal and throat swab specimens were the least likely to transmit infection, and the combination gave the highest yield for coronavirus detection by RT-PCR. Positive virologic test results in patient groups were associated with mechanical ventilation or death (p < 0.001), suggesting a correlation between viral load and disease severity.