Abattoirs ; manpower ; Adult ; Antibodies, Viral ; blood ; China ; Humans ; Influenza A Virus, H1N1 Subtype ; immunology ; Influenza A Virus, H3N2 Subtype ; immunology ; Influenza A Virus, H3N8 Subtype ; immunology ; Influenza A Virus, H5N1 Subtype ; immunology ; Influenza A Virus, H9N2 Subtype ; immunology ; Influenza A virus ; classification ; immunology ; Influenza, Human ; epidemiology ; prevention & control ; virology ; Middle Aged ; Population Surveillance ; methods ; Seroepidemiologic Studies ; Young Adult

Animals ; Birds ; China ; epidemiology ; Humans ; Influenza A Virus, H5N1 Subtype ; physiology ; Influenza A Virus, H7N7 Subtype ; physiology ; Influenza A Virus, H9N2 Subtype ; physiology ; Influenza A virus ; physiology ; Influenza, Human ; epidemiology ; prevention & control

The expression of the hemagglutinins of Avian influenza virus H5 H7and H9 subtypes was studied in this article by Pichia pastoris, one of the eukaryotis expression systems. Three reconstructed expression plasmids and engineering strains, named pPIC9K-H5HA, pPIC9K-H7HA, pPIC9K-H9HA and GS115/pPIC9K-H5HA, GS115/pPIC9K-H7HA, GS115/pPIC9K-H9HA repectively, were obtained. The reconstructed yeast engineering strains were identified by MD and MM plate selecting and PCR. The induced interests proteins were examined by SDS-PAGE and Western-bloting,the results showed that the interest genes were expressed exactly. And this will be helpful in the future study of antigen detection and antibody detection kit, as well in the subunit vaccines developing.
Animals ; Hemagglutinin Glycoproteins, Influenza Virus ; biosynthesis ; genetics ; Influenza A Virus, H5N1 Subtype ; genetics ; Influenza A Virus, H7N7 Subtype ; genetics ; Influenza A Virus, H9N2 Subtype ; genetics ; Pichia ; genetics ; metabolism

Specific primers and TaqMan MGB probes were designed with Primer Express 2.0 software according to the conserved region of the H5, H9, H7 subtype AIV hemagglutinin gene to make research of real-time fluorescent one-step PCR in the differential detection of H5, H9, H7 subtype avian influenza inactivated vaccines. The result showed that the method was specific and reproducible. No cross-reaction was discovered with other avian disease vaccines. Real-time fluorescent PCR provided a specific, sensitive, rapid and convenient method for the subtype identification of avian influenza inactivated vaccines.
Animals ; Hemagglutinin Glycoproteins, Influenza Virus ; immunology ; Humans ; Influenza A Virus, H5N1 Subtype ; immunology ; Influenza A Virus, H7N7 Subtype ; immunology ; Influenza A Virus, H9N2 Subtype ; immunology ; Influenza A virus ; classification ; immunology ; Influenza Vaccines ; analysis ; classification ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Vaccines, Inactivated ; analysis

In 2013, two episodes of influenza emerged in China and caused worldwide concern. A new H7N9 avian influenza virus (AIV) first appeared in China on February 19, 2013. By August 31, 2013, the virus had spread to ten provinces and two metropolitan cities. Of 134 patients with H7N9 influenza, 45 died. From then on, epidemics emerged sporadically in China and resulted in several victims. On November 30, 2013, a 73-year-old woman presented with an influenza-like illness. She developed multiple organ failure and died 9 d after the onset of disease. A novel reassortant AIV, H10N8, was isolated from a tracheal aspirate specimen that was obtained from the patient 7 d after onset. This case was the first human case of influenza A subtype H10N8. On 4 February, 2014, another death due to H10N8 avian influenza was reported in Jiangxi Province, China.
Aged ; China ; epidemiology ; Female ; Humans ; Influenza A Virus, H10N8 Subtype ; classification ; Influenza A Virus, H7N9 Subtype ; classification ; Influenza A Virus, H9N2 Subtype ; classification ; Influenza, Human ; epidemiology ; virology ; Phylogeny ; Reassortant Viruses ; classification

BACKGROUND: The new emerging avian influenza A H7N9 virus, causing severe human infection with a mortality rate of around 41%. This study aims to provide a novel treatment option for the prevention and control of H7N9. METHODS: H7 hemagglutinin (HA)-specific B cells were isolated from peripheral blood plasma cells of the patients previously infected by H7N9 in Jiangsu Province, China. The human monoclonal antibodies (mAbs) were generated by amplification and cloning of these HA-specific B cells. First, all human mAbs were screened for binding activity by enzyme-linked immunosorbent assay. Then, those mAbs, exhibiting potent affinity to recognize H7 HAs were further evaluated by hemagglutination-inhibiting (HAI) and microneutralization in vitro assays. Finally, the lead mAb candidate was selected and tested against the lethal challenge of the H7N9 virus using murine models. RESULTS: The mAb 6-137 was able to recognize a panel of H7 HAs with high affinity but not HA of other subtypes, including H1N1 and H3N2. The mAb 6-137 can efficiently inhibit the HA activity in the inactivated H7N9 virus and neutralize 100 tissue culture infectious dose 50 (TCID50) of H7N9 virus (influenza A/Nanjing/1/2013) in vitro, with neutralizing activity as low as 78 ng/mL. In addition, the mAb 6-137 protected the mice against the lethal challenge of H7N9 prophylactically and therapeutically. CONCLUSION The mAb 6-137 could be an effective antibody as a prophylactic or therapeutic biological treatment for the H7N9 exposure or infection.
Animals ; Antibodies, Monoclonal/therapeutic use* ; Antibodies, Neutralizing/therapeutic use* ; Antibodies, Viral ; Hemagglutinins ; Humans ; Influenza A Virus, H1N1 Subtype ; Influenza A Virus, H3N2 Subtype ; Influenza A Virus, H7N9 Subtype ; Influenza Vaccines ; Influenza in Birds ; Influenza, Human/prevention & control* ; Mice

OBJECTIVETo establish a multiplex RT-PCR-based reverse dot blot hybridization technique to detect influenza viruses.

METHODSObtain the HA nucleotide sequences of seasonal influenza H1N1, seasonal influenza H3N2, influenza H1N1 and human avian influenza H5N1 from GenBank. Design primers in conservative district and probes t in high variable region respectively, after analyzing the HA nucleotide sequences of influenza virus through the Vector NTI 9.0. Establish and optimize multiple RT-PCR system by comparing amplification efficiency and specificity at different primer concentrations. Establish the reverse dot hybridization system after optimizing the concentration of probes. To compare the sensitivity and specificity of this technique and the general RT-PCR Method through extracting the viral RNA of the mentioned influenza virus which are to be the reference substance.

RESULTSSuccessfully establish a multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses. This technique is 100-1000 times more sensitive than gel electrophoresis method, and it has a good specificity.

CONCLUSIONSuccessfully established multiplex RT-PCR-based reverse dot blot hybridization technique for detecting influenza viruses.

Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; Influenza A Virus, H3N2 Subtype ; genetics ; isolation & purification ; Influenza A Virus, H5N1 Subtype ; genetics ; isolation & purification ; Influenza, Human ; diagnosis ; virology ; Nucleic Acid Hybridization ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

Disease Outbreaks ; Humans ; Influenza A Virus, H1N1 Subtype ; physiology ; Influenza A Virus, H5N1 Subtype ; physiology ; Influenza, Human ; epidemiology ; prevention & control ; transmission ; virology

To examine the phylogenetic information regarding the gene pool of AIV in domestic ducks in eastern China, the NA genes of twenty-six viruses isolated during 2002-2006, including two H1N1 strains, tenH3N1 strains and fourteen HSN1 strains, which reflected the predominant N1 subtype viruses were subjected to phylogenetic analysis. The results indicated that AIVs of N1 subtype circulating in domestic ducks in eastern China were undergoing a gradual evolution. Analysis of the deduced amino acid sequences revealed that NAs from all isolated H5N1 viruses had a 20-aa deletion in the stalk region (residues 49-68), whereas no deletion was seen in the NAs from other HA subtype viruses. The viruses of H3N1 and H1N1 might have a propensity for reassortment of NA genes, whereas no direct evidence of reassortment of NA gene was obtained in H5N1 viruses.
Animals ; Birds ; China ; DNA, Viral ; analysis ; Evolution, Molecular ; Humans ; Influenza A Virus, H1N1 Subtype ; classification ; genetics ; Influenza A Virus, H3N2 Subtype ; classification ; genetics ; Influenza A Virus, H5N1 Subtype ; classification ; enzymology ; genetics ; Influenza A virus ; classification ; enzymology ; genetics ; Influenza in Birds ; virology ; Influenza, Human ; virology ; Neuraminidase ; genetics ; Phylogeny ; Poultry Diseases ; virology ; Sequence Alignment ; Sequence Deletion

OBJECTIVE: 1.To evaluates and recognizes findings in chest radiograph in patients with laboratory-confirmed S-OIV (H1N1) infection treated at UKMMC. 2. To evaluate whether the findings on initial chest radiographs of influenza A (H1N1) patients can help to predict the prognosis. MATERIAL AND METHODS: Total of 109 adult patients presenting to the Universiti Kebangsaan Malaysia Medical Centre (UKMMC) with flu-like symptoms who were positive for influenza A and these patients had underwent chest radiographs (CXR). The initial CXRs were evaluated for the pattern (consolidation, ground-glass, and reticulation), distribution, and extend of abnormality. The disease is classifies by the clinical severity (mild, moderate or severe illness) and adverse outcome (ventilated, death or recovered well). RESULTS: The initial CXRs were normal in 56% of cases. The predominant radiographic finding was consolidation, most commonly involving the middle and lower zones (35% of cases). There is no significant association between initial CXR findings with the patient clinical outcome either fully recovered or death/ ventilated. CONCLUSION: Normal chest radiographs is the most common radiographic finding in S-OIV (H1N1) infection and the most common abnormal lung finding is consolidation. Initial chest radiographs did not determine the patient clinical outcome and a normal initial radiograph could not exclude adverse outcome.
Influenza A Virus, H1N1 Subtype