Long-term passage of duck Tembusu virus in BHK-21 cells generates a completely attenuated and immunogenic population with increased genetic diversity
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Keywords:
Flavivirus
Attenuated vaccine
Serial passage
Reverse Genetics
Japanese encephalitis vaccine
Serial passage
Attenuated vaccine
Vero cell
Reversion
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A total of 1,136 samples from 289 households in four provinces in northern Laos were subjected to Japanese encephalitis virus (JEV) and dengue virus hemagglutination inhibition (DENV HI). Overall, antibodies to JEV were detected by HI in 620 (54.6%) of 1,136 people; of which 217 (19.1%) had HI activity against JEV only. Antibodies to DENV4 were detected by HI in 526 (46.3%) of 1,136 people; of which 124 (10.9%) had HI activity against DENV4 only. Antibodies to DENV1–3 were detected by HI in 296 (26.1%), 274 (24.1%), and 283 (24.9) of 1,136 people, respectively; of which 7, 1, and 0, respectively, had HI activity against DENV1–3 only. JEV was the most prevalent Flavivirus in Oudomxay, Luangprabang, and Huaphan provinces and DENV4 was the most prevalent in Xiengkhouang province. Seroprevalence for JEV increased with increasing age and wealth and was higher in villages where rice was cultivated in paddy fields and highest for people of Lao-Tai ethnicity.
Seroprevalence
Flavivirus
Hemagglutination assay
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Flavivirus
Dengue vaccine
Attenuated vaccine
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We have generated new influenza A virus live attenuated vaccine candidates by site-directed mutagenesis and reverse genetics. By mutating specific amino acids in the PB2 polymerase subunit, two temperature-sensitive (ts) attenuated viruses were obtained. Both candidates have 38 degrees C shutoff temperatures in MDCK cells, are attenuated in the respiratory tracts of mice and ferrets, and have very low reactogenicity in ferrets. Infection of mice or ferrets with either mutant conferred significant protection from challenge with the homologous wild-type virus. Three tests for genetic stability were used to assess the propensity for reversion to virulence: 14 days of replication in nude mice, growth at 37 degrees C in tissue culture, and serial passage in ferrets. One candidate, which contains mutations intended to reduce the ability of PB2 to bind to cap structures, was stable in all three assays, whereas the second candidate, which contains mutations found only in other ts strains of influenza virus, lost its ts phenotype in the last two assays. This approach has therefore enabled the creation of live attenuated influenza A virus vaccine candidates suitable for human testing.
Attenuated vaccine
Reverse Genetics
Serial passage
Reactogenicity
Reversion
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日本脑炎病毒(JEV ) 与包括黄发烧病毒，登革热病毒和韦斯特尼罗河病毒的人的病原体有关仔细是忍受蚊子的 flavivirus。当前为所有 flavivirus 没有有效抗病毒的治疗，仅仅一些高度有效的疫苗为人的使用被准许。在这份报纸，六 heterologous flaviviruses (DENV1-4， WNV 和 JEV ) 的 E 蛋白质领域 III (DIII ) 成功地在 Escherichia coli 被表示。蛋白质在增溶和 refolding 过程以后被净化，由 SDS 页并且西方的弄污描绘了。竞争抑制证明所有 recombinant flavivirus DIII 蛋白质堵住了 JEV 的入口进 BHK-21 房间。进一步的研究显示可溶的 recombinant flavivirus DIII 导致的抗体部分保护了老鼠免于致命的 JEV 挑战。这些结果证明 recombinant flavivirus DIII 蛋白质能竞争地禁止 JEV 感染，并且有合适的合拢的 flavivirus DIII 的免疫导致了对在老鼠的 JEV 感染跨保护，暗示 DIII 的一个可能的角色为在象它在为在动物模型的免疫的抗原的使用一样的 flavivirus 之中跨保护。
Flavivirus
Heterologous
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Staphylococcus aureus rich in protein A when coated with monoclonal antibodies (MoAb) to Japanese encephalitis virus (JEV) gave a highly specific reaction with flavivirus antigens. The bacteria coated with JEV species-specific MoAb gave a strong co-agglutination with fifty-six JEV isolates from various parts of China, but no co-agglutination with Murray Valley encephalitis (MVE) and Kunjin (Kun) virus antigens. The flavivirus- and subgroup-specific MoAbs were reactive with MVE and Kun, as well as with the majority of the JEV strains. Blocking test with homologous MoAbs abolished co-agglutination further confirming its specificity. Numerous virus particles were observed on the surface of MoAb-coated staphylococci under the electron microscope after co-agglutination. The test appeared rapid, specific, simple to perform, and useful for rapid detection and identification of flaviviruses.
Flavivirus
Agglutination (biology)
Direct agglutination test
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Flavivirus
Heterologous
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Objective To evaluate the Flavivirus specific monoclonal antibody(McAb) 2A10 as detective antibody for simultaneously identify tick borne encephalitis virus( TBEV), Japanese encephalitis virus( JEV), dengue ( DEN )-2, DEN-4 and yellow fever virus ( YFV ) by antibody microarray technique.Methods The antibody microarray was developed by spotting TBEV, JEV, DEN-2, DEN-4 and YFV specific McAb on chip as capture antibodies. After incubating with cultured viral supernatants of the above viruses, CY3 labeled detective antibody 2A10 was added to the chips. After reaction, the antibody microarray was scanned and the results were analyzed. By comparing the signal intensities of different spots on chips,the detecting titre and sensitivity of 2A10 for Flavivirus were determined, and the value of 2A10 in detection of Flavivirus was evaluated. Results The hybridization results demonstrated that the titre of 2A10 for Flavi2A10 was specific for Flavivirus and could be used as universal detective antibody for Flavivirus on antibody microarray.
Key words:
Protein chip; High-flux; Flavivirus; Monoclonal antibody
Flavivirus
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The genus Flavivirus includes several pathogenic agents that cause severe illness in humans. Re-emergence of West Nile virus in Europe and continuous spread of certain flaviviruses such as dengue, yellow fever and Japanese encephalitis viruses represent a global danger to public health. Therefore, a rapid and accurate molecular method is required for diagnostics and epidemiological surveillance of flaviviruses.A Pan-Flavi quantitative RT-PCR assay using a Locked-Nucleic Acid probe targeting the flavivirus NS5 gene was developed and optimized to detect a wide range of flaviviruses simultaneously. The specificity and sensitivity of the Pan-Flavi assay were tested using RNA of different flaviviruses and non-flaviviruses. Furthermore, the assay was compared directly to flavivirus species-specific assays for the ability to detect flaviviruses sensitively.Two degenerate primers and one Locked-Nucleic Acids probe were designed to amplify most of the flaviviruses. To increase the specificity and fluorescence signal of the Pan-Flavi assay for detection of yellow fever virus and dengue virus 4, additional primers and probes were included. Viral RNA of thirty different flaviviruses was detected, verifying the broad range specificity. The testing of this assay was successful, using standard plasmid and RNA dilutions of yellow fever virus vaccine strain, dengue virus 1 and tick-borne encephalitis virus, with a sensitivity limit of 10-100 genome copies/reaction. Also comparatively good results were achieved for detecting different flaviviruses by the Pan-Flavi assay when compared to the flavivirus species-specific assays.The assay is rapid, broad-range flavivirus-specific and highly sensitive making it a valuable tool for rapid detection of flaviviruses in livestock samples, epidemiological studies or as useful complement to single flavivirus-specific assays for clinical diagnosis.
Flavivirus
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The genus Flavivirus of the family Flaviviridae comprises over 70 viruses,
many of which, such as the Dengue (DEN) virus, Japanese Encephalitis (JE) virus,
West Nile virus (WNV), St. Louis Encephalitis (SLE) virus, and Yellow fever (YF)
virus are important human pathogens. Flavivirus can be transmitted to humans via
either a mosquito or tick vector. The flavivirus genera are of importance to the
medical community because they have been found to be the causative agent of many
endemic and epidemic illnesses across the world. Japanese Encephalitis virus (JEV)
is the most important cause of viral encephalitis in Asia based on its frequency and
severity. With the near eradication of poliomyelitis, JEV is now the leading cause of
childhood viral neurological infection and disability in Asia.
Flavivirus
Veterinary virology
Viral encephalitis
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