POULTRY SCIENCE
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2024
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IF 3.8
Sha-Sha Chen,Yong-Lei Yang,Hong-Yun Wang,Tian-Kui Guo,Riaz-M Azeem,Chun-Wei Shi,Gui-Lian Yang,Hai-Bin Huang,Yan-Long Jiang,Jian-Zhong Wang,Xin Cao,Nan Wang,Yan Zeng,Wen-Tao Yang,Chun-Feng Wang
ABSTRACT Avian influenza virus (AIV) subtype H9N2 has significantly threatened the poultry business in recent years by having become the predominant subtype in flocks of chickens, ducks, and pigeons. In addition, the public health aspects of H9N2 AIV pose a significant threat to humans. Early and rapid diagnosis of H9N2 AIV is therefore of great importance. In this study, a new method for the detection of H9N2 AIV based on fluorescence intensity was successfully established using CRISPR/Cas13a technology. The Cas13a protein was first expressed in a prokaryotic system and purified using nickel ion affinity chromatography, resulting in a high-purity Cas13a protein. The best RPA(Recombinase polymerase amplification) primer pairs and crRNA were designed and screened, successfully constructing the detection of H9N2 AIV based on CRISPR/Cas13a technology. Optimal concentration of Cas13a and crRNA was determined to optimise the constructed assay. The sensitivity of the optimised detection system is excellent, with a minimum detection limit of 10° copies/μL and didn't react with other avian susceptible viruses, with excellent specificity. The detection method provides the basis for the field detection of the H9N2 AIV.
研究领域:
分子生物学
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病毒学
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生物技术
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公共卫生
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产品关联
DOI
PubMed
高影响因子
ACS Applied Materials & Interfaces
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2019
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IF 8.456
Xiaolun Peng,Guan Luo,Zhen Wu,Wei Wen,Xiuhua Zhang,Shengfu Wang
The outbreak of H9N2 avian influenza virus (H9N2 AIV) brings high mortality and huge economic losses every year. Sensitive and reliable detection methods are essential to timely diagnosis and treatment. Herein, a dual-modality immunoassay is proposed for H9N2 AIV detection by employing fluorescent-magnetic-catalytic nanospheres (FMCNs) as labels and alkaline phosphatase (ALP)-induced metallization as a signal amplification strategy. The excellent magnetic properties of FMCNs endow the assay a potential application in complex samples. Also, the excellent fluorescence properties of FMCNs enable fluorescence modality readout. The antibodies on the FMCN surface can achieve efficient capture and separation of targets. Amplified electrochemical modality readout can be obtained through ALP-catalyzed silver deposition. Dual-modality immunoassay combined the advantages of electrochemical assay with fluorescence assay and provides accurate detection results to meet different testing needs. With two quantitative analysis forms, H9N2 AIV can be detected by electrochemical signals with a quantitation range of 0.1 to 1000 ng/mL and a detection limit of 10 pg/mL. The linear range is 300 to 1000 ng/mL with a detection limit of 69.8 ng/mL by the fluorescence signal readout. Moreover, the specificity, anti-interference ability, accuracy, and diversity of the proposal have unlimited potential for early diagnosis of suspect infections.
研究领域:
病毒学
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生物医学工程
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分析化学
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2019
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IF 5.256
Guangyang Xiao,Beibei Chen,Man He,Bin Hu
Avian influenza virus (AIVs), hosted in poultry, are the pathogens of many poultry diseases and human infections, which bring huge losses to the poultry breeding industry and huge panic to society. Therefore, it is of great significance to establish accurate and sensitive detection methods for AIVs. In this work, a dual-mode detection method based on immuno-rolling circle amplification (immuno-RCA) and quantum dots (QDs) labeling for inductively coupled plasma mass spectrometry (ICP-MS) and fluorescence detection of H9N2 AIV was developed. The dual-mode detection of the QDs by ICP-MS and fluorescence is used to achieve mutual verification within the analysis results, thus improving the accuracy of the method. With the immuno-RCA, the sensitivity of the method was increased by two orders of magnitude. The limit of detection of the proposed method is 17 ng L −1 and 61 ng L −1 , and the linear range of the proposed method is 0.05–5 ng mL −1 and 0.1–5 ng mL −1 with ICP-MS and fluorescence detection, respectively. The relative standard deviation (n = 7) is 4.9% with ICP-MS detection and 3.1% with fluorescence detection. Furthermore, the proposed method was applied to the analysis of chicken serum samples, no significant different was found for two modes detection and the recoveries of the spiking experiments are acceptable, indicating that the method has good practical potential for real sample analysis.
研究领域:
分析化学
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病毒学
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生物技术
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DOI
PubMed
高影响因子
ANALYTICA CHIMICA ACTA
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2021
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IF 6.558
Bin Wang,Baojie Li,Huachuan Huang,Shuwei Yang,Dan Jian,Jing Liu,Keding Yan,Yanke Shan,Shouyu Wang,Fei Liu
Avian influenza virus (AIV) is a serious zoonotic disease causing severe damages to both poultry industry and human health. To rapidly detect AIV on-site with high sensitivity and accuracy, we design sensitive antibody fluorescence immunosorbent assay (SAFIA) on AIV H9N2 antibody. In SAFIA, hemagglutinin (HA1) protein coated sample chamber specifically binds targets but remarkably reduces non-specific absorption; Protein L coated polystyrene microsphere captures target through secondary antibody to significantly amplify the fluorescence signal; and a portable fluorescence counter automatically measures the fluorescence spot density for AIV H9N2 antibody detection. Proved by practical applications, SAFIA could probe AIV H9N2 antibody in high sensitivity and selectivity , and distinguish positive and negative serum samples in high accuracy. Additionally, SAFIA can rapidly detect AIV H9N2 antibody at room temperature only requiring simple operations as well as cost-effective and compact devices. Therefore, SAFIA is a potential new-generation tool in rapid on-site testing for agricultures.
研究领域:
兽医学
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分子生物学
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诊断技术
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动物健康
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DOI
PubMed
高影响因子
MICROBIAL PATHOGENESIS
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2024
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IF 3.3
Tongyu Zhang,Yawen Tian,Xiao Zhang,Wenfeng Wang,Yingkai He,Chongbo Ge,Futing Jia,Zhannan Wang,Yanlong Jiang
The H9N2 avian influenza virus (AIV) is spreading worldwide. Presence of H9N2 virus tends to increase the chances of infection with other pathogens which can lead to more serious economic losses. In a previous study, a regulated delayed lysis Salmonella vector was used to deliver a DNA vaccine named pYL233 encoding M1 protein, mosaic HA protein and chicken GM-CSF adjuvant. To further increase its efficiency, chitosan as a natural adjuvant was applied in this study. The purified plasmid pYL233 was coated with chitosan to form a DNA containing nanoparticles (named CS233) by ionic gel method and immunized by intranasal boost immunization in birds primed by oral administration with Salmonella strain. The CS233 DNA nanoparticle has a particle size of about 150 nm, with an encapsulation efficiency of 93.2 ± 0.12 % which protected the DNA plasmid from DNase I digestion and could be stable for a period of time at 37°. After intranasal boost immunization, the CS233 immunized chickens elicited higher antibody response, elevated CD4 + T cells and CD8 + T cells activation and increased T-lymphocyte proliferation, as well as increased productions of IL-4 and IFN-γ. After challenge, chickens immunized with CS233 resulted in the lowest levels of pulmonary virus titer and viral shedding as compared to the other challenge groups. The results showed that the combination of intranasal immunization with chitosan-coated DNA vaccine and oral immunization with regulatory delayed lytic Salmonella strain could enhance the immune response and able to provide protection against H9N2 challenge.
研究领域:
免疫学
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病毒学
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兽医学
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DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
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2022
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IF 12.545
Jinrong Liu,Chong Ma,Siwei Shi,Heng Liu,Wei Wen,Xiuhua Zhang,Zhen Wu,Shengfu Wang
As an important component of the COVID-19 mRNA vaccines, liposomes play a key role in the efficient protection and delivery of mRNA to cells. Herein, due to the controllable release amplification strategy of liposomes, a reliable and robust single-particle collision electrochemical (SPCE) biosensor was constructed for H9N2 avian influenza virus (H9N2 AIV) detection by combining liposome encapsulation-release strategy with immunomagnetic separation. The liposomes modified with biotin and loaded with platinum nanoparticles (Pt NPs) were used as signal probes for the first time. Biotin facilitated the coupling of biomolecules (DNA or antibodies) through the specific reaction of biotin-streptavidin. Each liposome can encapsulate multiple Pt NPs, which were ruptured under the presence of 1 × PBST (phosphate buffer saline with 0.05% Tween-20) within 2 min, and the encapsulated Pt NPs were released for SPCE experiment. The combination of immunomagnetic separation not only improved the anti-interference capabilities but also avoided the agglomeration of Pt NPs, enabling the SPCE biosensor to realize ultrasensitive detection of 18.1 fg/mL H9N2 AIV. Furthermore, the reliable SPCE biosensor was successfully applied in specific detection of H9N2 AIV in complex samples (chicken serum, chicken liver and chicken lung), which promoted the universality of SPCE biosensor and its application prospect in early diagnosis of diseases.
研究领域:
生物化学
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分析化学
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病毒学
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生物医学工程
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DOI
PubMed
高影响因子
SPECTROCHIMICA ACTA PART B-ATOMIC SPECTROSCOPY
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2017
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IF 3.241
Guangyang Xiao,Beibei Chen,Man He,Kaiwen Shi,Xing Zhang,Xiaoting Li,Qiumei Wu,Daiwen Pang,Bin Hu
Avian influenza viruses are the pathogens of global poultry epidemics, and may even cause the human infections. Here, we proposed a novel inductively coupled plasma mass spectrometry (ICP-MS) based immunoassay with gold nanoparticles (Au NPs) labeling for the determination of H9N2 virions. Magnetic-beads modified with anti-influenza A H9N2 hemagglutinin mono-antibody (mAb-HA) were utilized for the capture of H9N2 virions in complex matrix; and Au NPs conjugated with mAb-HA were employed for the specific labeling of H9N2 virions for subsequent ICP-MS detection. With a sandwich immunoassay strategy, this method exhibited a high specificity for H9N2 among other influenza A virions such as H1N1 and H3N2. Under the optimized conditions, this method could detect as low as 0.63 ng mL − 1 H9N2 virions with the linear range of 2–400 ng mL − 1 , the relative standard deviation for seven replicate detections of H9N2 virions was 7.2% (c = 10 ng mL − 1 ). The developed method was applied for the detection of H9N2 virions in real-world chicken dung samples, and the recovery for the spiking samples was 91.4–116.9%. This method is simple, rapid, sensitive, selective, reliable and has a good application potential for virions detection in real-world samples.
研究领域:
病毒学
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分析化学
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生物技术
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环境科学
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DOI
高影响因子
危险品化学品经营许可证（带存储）