RESUMO
Two pairs of primers were designed to bind conserved genomic regions of goose parvovirus (GPV) and goose astrovirus (GAstV) to establish a simple, sensitive, and highly specific duplex quantitative PCR (qPCR) method to simultaneously detect the two viruses. The duplex qPCR can distinguish GPV (melting point: 82.1 °C) and GAstV (melting point: 79.8 °C) by the peaks of their individual melting curves. Mixed testing with other waterfowl viruses produced no nonspecific peaks. The established standard curves showed good linear relationships (R2 > 0.997) and the limits of detection (LOD) for GPV and GAstV were 5.74 × 101 and 6.58 × 101 copies/µL, respectively. Both intra- and inter-assay coefficients of variation were <2%, indicating that the method has good repeatability. Twenty tissue samples from diseased geese were examined with the duplex qPCR assay and conventional PCR. Duplex qPCR showed positive rates of 25% for GPV and 45% for GAstV, and the positive rate for GPV and GAstV coinfection was 15%, slightly higher than the results for conventional PCR. These results indicated that this duplex qPCR method is highly sensitive, specific, and reproducible, and is suitable for epidemiological studies to effectively control the transmission of GPV and GAstV.
Assuntos
Infecções por Astroviridae/diagnóstico , Infecções por Astroviridae/veterinária , Avastrovirus/isolamento & purificação , Benzotiazóis/metabolismo , Diaminas/metabolismo , Infecções por Parvoviridae/diagnóstico , Infecções por Parvoviridae/veterinária , Parvovirinae/isolamento & purificação , Quinolinas/metabolismo , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Gansos/virologia , Padrões de Referência , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
Goose circovirus (GoCV) is a potential immunosuppressive virus that poses a great hazard to the goose industry and has been shown to be widely distributed throughout China. We have established a fast, sensitive and highly specific TaqMan real-time quantitative PCR detection method for this virus. Specific primers and probes were designed against the conserved regions of the genomic GoCV Rep gene. The results showed that the assay was highly specific and sensitive for GoCV and did not cross-react with other non-targeted waterfowl viruses. The established method will be helpful for epidemiological detection and may be effective in the prevention and control of the disease.
Assuntos
Circovirus/genética , Circovirus/isolamento & purificação , Reação em Cadeia da Polimerase em Tempo Real/métodos , Animais , Bioensaio , Gansos/virologia , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: SRY-related HMG box-12, which is associated with the prognosis of cancer, has been frequently described. However, both SRY-related HMG box-12 expression and its relationship with clinicopathological variables and patient survival have not been defined in gastric cancer. The aim of our study was to examine the prognostic value of SRY-related HMG box-12 expression in patients with gastric cancer. METHODS: In this study, we determined SRY-related HMG box-12 expression in 79 primary gastric cancer tissues and 79 matched adjacent nontumor tissues by immunohistochemistry and then calculated the survival rate using the Kaplan-Meier method. Cox proportional hazard regression model was used to analyze predictors of gastric cancer. Western blot and quantitative real-time polymerase chain reaction were used to investigate the difference in SRY-related HMG box-12 expression between normal gastric epithelial cells and gastric cancer cells at the protein level and RNA level, respectively. RESULTS: SRY-related HMG box-12 was downregulated in gastric cancer tissues. Low SRY-related HMG box-12 expression was significantly associated not only with lymph node metastasis (P = .027) and TNM stage (P = .021) but also with disease-specific survival in patients with gastric cancer. Multivariate analysis demonstrated TNM stage was an independent factor predicting poor survival (P = .034). CONCLUSIONS: Low SRY-related HMG box-12 expression is associated with poor clinical outcomes in gastric cancer.
Assuntos
Biomarcadores Tumorais/metabolismo , Regulação Neoplásica da Expressão Gênica , Fatores de Transcrição SOXC/metabolismo , Neoplasias Gástricas/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Linhagem Celular Tumoral , Células Cultivadas , Feminino , Humanos , Estimativa de Kaplan-Meier , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Prognóstico , Neoplasias Gástricas/mortalidade , Neoplasias Gástricas/patologia , Taxa de SobrevidaRESUMO
Porcine parvovirus 7 (PPV7), a new serotype of the porcine parvovirus, was discovered in swine of the USA in 2016. Recently, PPV7 was detected in Anhui province, China. Twenty-four of the 120 lung samples were PPV7-positive. Three PPV7 strains were sequenced and named PPV7/China/AHbz, PPV7/China/AHhf, and PPV7/China/AHmas, respectively. The complete genome and NS1 gene nucleotides of the three PPV7 strains showed 80.0%-98.4% and 94.4%-98.7% sequence identity, respectively, to the other PPV7 strains obtained from NCBI. The three PPV7 strains from Anhui share a common origin with a PPV7 GX49 strain isolated in Guangxi. These results help to understand the molecular epidemiology of PPV7.
