ABSTRACT
BACKGROUND: Human adenoviruses are a common group of viruses that cause acute infectious diseases. Human adenovirus (HAdV) 3 and HAdV 7 cause major outbreaks of severe pneumonia. A reliable and practical method for HAdV typing in clinical laboratories is lacking. A simple, rapid and accurate molecular typing method for HAdV may facilitate clinical diagnosis and epidemiological control. METHODS: We developed and evaluated duplex real-time recombinase-aided amplification (RAA) assays incorporating competitive internal controls for detection of HAdV 3 and HAdV 7, respectively. The assays were performed in a one-step in a single tube reaction at 39° for 20 min. RESULTS: The analytical sensitivities of the duplex RAA assays for HAdV 3 and HAdV 7 were 5.0 and 14.8 copies per reaction, respectively (at 95% probability by probit regression analysis). No cross-reaction was observed with other types of HAdV or other common respiratory viruses. The duplex RAA assays were used to detect 152 previously-defined HAdV-positive samples. These results agreed with those obtained using a published triplex quantitative real-time PCR protocol. CONCLUSIONS: We provide the first report of internally-controlled duplex RAA assays for the detection of HAdV 3 and HAdV 7. These assays effectively reduce the rate of false negative results and may be valuable for detection of HAdV 3 and HAdV 7 in clinical laboratories, especially in resource-poor settings.
Subject(s)
Adenoviruses, Human/isolation & purification , Nucleic Acid Amplification Techniques/methods , Nucleic Acid Amplification Techniques/standards , Recombinases/genetics , Adenoviruses, Human/genetics , Humans , Real-Time Polymerase Chain Reaction/methods , Real-Time Polymerase Chain Reaction/standards , Respiratory Tract Infections/epidemiology , Sensitivity and Specificity , Serogroup , TemperatureABSTRACT
OBJECTIVE: To be acquainted with genetic characteristics and variation of mumps virus strains circulating in Hunan province. METHODS: Mumps virus (MV) strains were isolated using Vero/ SLAM cells. The small hydrophobic protein (SH) genes of MV isolates were sequenced, and the sequences were analysed phylogenetically between the isolated strains and other reference mumps strains. RESULTS: 4 mumps virus strains were isolated from 16 specimens collected in 2011 from different regions of Hunan province. The genotype of isolated strains were supposed to be F type. CONCLUSION: Genotype F is the main genotype of circulating strains in Hunan province in 2011 and there is no variation between genotype.
Subject(s)
Mumps virus/genetics , Mumps/virology , Animals , China/epidemiology , Chlorocebus aethiops , Genetic Variation , Genotype , Humans , Mumps/epidemiology , Mumps virus/isolation & purification , Phylogeny , Vero Cells , Viral Proteins/geneticsABSTRACT
OBJECTIVE: To study the global evolutionary characteristics of hemagglutinin gene HA1 of influenza H1N1 infecting different species during 2000-2009. METHODS: The target sequences were downloaded from NCBI and analyzed using bioinformatic software to construct the phylogenetic tree. RESULTS: The HA1 amino acid sequences of influenza H1N1 contained four mutated antigenic sites and receptor-binding sites, and the novel influenza virus shared most of the mutated amino acid sites with swine H1N1 influenza virus. CONCLUSION: The HA1 gene of novel influenza virus might originate from the early swine H1N1 influenza virus from North America, and in the evolutionary process, a number of important sites of HA1 gene mutated to result in the outbreak of influenza.
