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34 results on '"Yu, Kangzhen"'

12. Rapid detection of newly isolated Tembusu-related Flavivirus by reverse-transcription loop-mediated isothermal amplification assay

Author

Wang Youling, Yuan Xiaoyuan, Li Yufeng, Yu Kexiang, Yang Jinxing, Xu Huaiying, Zhang Yuxia, Yu Kangzhen, Liao Ming, and Qin Zhuoming

Subjects
Tembusu-related Flavivirus, newly-isolated virus, RT-PCR, RT-LAMP, Infectious and parasitic diseases, RC109-216
Abstract

Abstract Background From April 2010 to January 2011, a severe new viral disease had devastated most duck-farming regions in China. This disease affected not only laying ducks but also meat ducks, causing huge economic losses for the poultry industry. The objective of this study is to develop a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for the detection of the new virus related to Tembusu-related Flavivirus. Results The RT-LAMP assay is very simple and rapid, and the amplification can be completed within 50 min under isothermal conditions at 63°C by a set of 6 primers targeting the E gene based on the sequences analysis of the newly isolated viruses and other closely related Flavivirus.The monitoring of gene amplification can also be visualized by using SYBR green I fluorescent dye. In addition, the RT-LAMP assay for newly isolated Tembusu-related Flavivirus showed higher sensitivity with an RNA detection-limit of 2 copies/μL compared with 190 copies/μL of the conventional RT-PCR method. The specificity was identified without cross reaction to other common avian pathogens. By screening a panel of clinical samples this method was more feasible in clinical settings and there was higher positive coincidence rate than conventional RT-PCR and virus isolation. Conclusion The RT-LAMP assay for newly isolated Tembusu-related Flavivirus is a valuable tool for the rapid and real-time detection not only in well-equipped laboratories but also in general conditions.

Published
2011
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15. Comparison of protective effects among H5 subtype avian influenza DNA vaccines with optimized HA gene and expressive vectors

Author

Jiang YongPing, Chen Hua-lan, Bu ZhiGao, Yu Kangzhen, Deng Guohua, Li Chengjun, and Zhang Hong-bo

Subjects
Biology, medicine.disease_cause, Precipitin, Virology, Influenza A virus subtype H5N1, Virus, DNA vaccination, Vaccination, medicine, biology.protein, Viral shedding, Antibody, Agronomy and Crop Science, Biotechnology, Specific-pathogen-free
Abstract

To evaluate the effects of H5 subtype avian influenza DNA vaccine with HA gene and expressive vector optimization, groups of 3-week-old specific pathogen free (SPF) chickens were inoculated intramuscularly with a single dose of 100 μg or 10 μg of pCIHA5, pCAGGHA5, pCIoptiHA5 or pCAGGoptiHA5 in 200 μl volume. Another group of chickens was injected with 200 μl phosphate-buffered saline (PBS) as controls. Sera were collected every week after vaccination to detect the HI and agar-gel precipitin (AGP) antibodies. Four weeks after the single vaccination, all chickens were challenged with 100LD50 of the highly pathogenic A/Goose/GuangDong/1/96(H5N1) [Gs/GD/1/96(H5N1)]. Oropharyngeal and cloacal swab specimens were collected from all chickens 3, 5 and 7 days after inoculation for titration of virus in eggs. Chickens were observed daily for 2 weeks for signs of disease and death. Results showed that in the groups vaccinated with 100 μg pCAGGoptiHA5 and pCAGGHA5 chickens were completely protected from virus challenge (no disease signs, no virus shedding and no deaths), while only partial protection occurred in chickens vaccinated with 100 μg pCIoptiHA5 (75%) and pCIHA5 (50%). In groups vaccinated with 10 μg pCAGGoptiHA5 and pCAGGHA5 chickens were protected from virus challenge (no disease signs and no deaths). In the group vaccinated with 10 μg pCIoptiHA5 the protection ratio was 75%, while chickens vaccinated with 10 μg pCIHA5 all died after challenge. Results indicated that codon usage optimization of the HA gene and β-actin promoter enhanced the protection efficiency of H5 subtype avian influenza DNA vaccine and the construction pCAGGoptiHA5 could protect chickens from lethal H5N1 virus challenge even at the low dose of 10 μg, implying the possibility for the commercialization of an avian influenza DNA vaccine in the near future.

Published
2007

24. The NS Segment of an H5N1 Highly Pathogenic Avian Influenza Virus (HPAIV) Is Sufficient To Alter Replication Efficiency, Cell Tropism, and Host Range of an H7N1 HPAIV

Author

Ma, Wenjun, primary, Brenner, Dominique, additional, Wang, Zhongfang, additional, Dauber, Bianca, additional, Ehrhardt, Christina, additional, Högner, Katrin, additional, Herold, Susanne, additional, Ludwig, Stephan, additional, Wolff, Thorsten, additional, Yu, Kangzhen, additional, Richt, Jürgen A., additional, Planz, Oliver, additional, and Pleschka, Stephan, additional

Published
2010
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27. Newcastle Disease Virus-Based Live Attenuated Vaccine Completely Protects Chickens and Mice from Lethal Challenge of Homologous and Heterologous H5N1 Avian Influenza Viruses

Author

Ge, Jinying, primary, Deng, Guohua, additional, Wen, Zhiyuan, additional, Tian, Guobing, additional, Wang, Yong, additional, Shi, Jianzhong, additional, Wang, Xijun, additional, Li, Yanbing, additional, Hu, Sen, additional, Jiang, Yongping, additional, Yang, Chinglai, additional, Yu, Kangzhen, additional, Bu, Zhigao, additional, and Chen, Hualan, additional

Published
2007
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34. The NS Segment of an H5N1 Highly Pathogenic Avian Influenza Virus (HPAIV) Is Sufficient To Alter Replication Efficiency, Cell Tropism, and Host Range of an H7N1 HPAIV.

Author

Wenjun Ma, Brenner, Dominique, Zhongfang Wang, Dauber, Bianca, Ehrhardt, Christina, Högner, Katrin, Herold, Susanne, Ludwig, Stephan, Wolff, Thorsten, Yu, Kangzhen, Richt, Jürgen A., Planz, Oliver, and Pleschka, Stephan

Subjects
*AVIAN influenza, *GENETICS, *CELL lines, *RESPIRATORY organs, *EPITHELIAL cells, *ANTIVIRAL agents
Abstract

A reassortant avian influenza virus (designated FPV NS GD), carrying the NS-segment of the highly pathogenic avian influenza virus (HPAIV) strain A/Goose/Guangdong/1/96 (GD; H5N1) in the genetic background of the HPAIV strain A/FPV/Rostock/34 (FPV; H7N1), was rescued by reverse genetics. Remarkably, in contrast to the recombinant wild-type FPV (rFPV), the reassortant virus was able to replicate more efficiently in different human cell lines and primary mouse epithelia cells without prior adaptation. Moreover, FPV NS GD caused disease and death in experimentally infected mice and was detected in mouse lungs; in contrast, rFPV was not able to replicate in mice effectively. These results indicated an altered host range and increased virulence. Furthermore FPV NS GD showed pronounced pathogenicity in chicken embryos. In an attempt to define the molecular basis for the apparent differences, we determined that NS1 proteins of the H5N1 and H7N1 strains bound the antiviral kinase PKR and the F2F3 domain of cleavage and polyadenylation specificity factor 30 (CPSF30) with comparable efficiencies in vitro. However, FPV NS GD infection resulted in (i) increased expression of NS1, (ii) faster and stronger PKR inhibition, and (iii) stronger beta interferon promoter inhibition than rFPV. Taken together, the results shed further light on the importance of the NS segment of an H5N1 strain for viral replication, molecular pathogenicity, and host range of HPAIVs and the possible consequences of a reassortment between naturally occurring H7 and H5 type HPAIVs. [ABSTRACT FROM AUTHOR]

Published
2010
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