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Subtyping Animal Influenza Virus with General Multiplex RT-PCR and Liquichip High Throughput (GMPLex) 认领 引用 被引量：8
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作者 Zhi-feng Qin Jie Sun +11 位作者 Ti-kang Lu Shao-ling Zeng Qun-yi Hua Qing-yan Ling Shu-kun Chen Jian-qiang Lv Cai-hong Zhang Bing Cheng Zhou-xi Ruan Ying-zuo Bi Joseph J Giambrone Hong-zhuan Wu 《Virologica Sinica》 CAS CSCD 2012年第2期120-131,共12页
This study developed a multiplex RT-PCR integrated with luminex technology to rapidly subtype simultaneously multiple influenza viruses. Primers and probes were designed to amplify NS and M genes of influenza A viruse... This study developed a multiplex RT-PCR integrated with luminex technology to rapidly subtype simultaneously multiple influenza viruses. Primers and probes were designed to amplify NS and M genes of influenza A viruses HA gene of ill, H3, H5, HT, H9 subtypes, and NA gene of the N1 and N2 subtypes. Universal super primers were introduced to establish a multiplex RT-PCR （GM RT-PCR）. It included three stages of RT-PCR amplification, and then the RT-PCR products were further tested by LiquiChip probe, combined to give an influenza virus （IV） rapid high throughput subtyping test, designated as GMPLex. The IV GMPLex rapid high throughput subtyping test presents the following features： high throughput, able to determine the subtypes of 9 target genes in H1, H3, H5, H7, H9, N1, and N2 subtypes of the influenza A virus at one time; rapid, completing the influenza subtyping within 6 hours; high specificity, ensured the specificity of the different subtypes by using two nested degenerate primers and one probe, no cross reaction occurring between the subtypes, no non-specific reactions with other pathogens and high sensitivity. When used separately to detect the product of single GM RT-PCR for single H5 or N1 gene, the GMPLex test showed a sensitivity of 10-5（= 280ELDs0） forboth tests and the Luminex qualitative ratio results were 3.08 and 3.12, respectively. When used to detect the product of GM RT-PCR for H5N1 strain at the same time, both showed a sensitivity of 10-4（=2800 ELD50）. The GMPLex rapid high throughput subtyping test can satisfy the needs of influenza rapid testing. 展开更多
关键词 Influenza Virus General multiplex RT-PCR Iuminex assay Subtyping HA and NA genes
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Designing Primers for H5 and H7 Subtypes of Avian Influenza Virus and Multiplex RT-PCR Amplification 认领 引用 被引量：7
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作者 张文慧 郭华 +2 位作者 王伟利 刘明 钱爱东 《Agricultural Science & Technology》 CAS 2008年第1期15-17,共3页
[Objective] The research aimed to design primers that are suitable for detecting H5 and H7 subtypes of avian influenza virus （AIV） ; [Method] DNAStar was used to analyze the homology of the sequences of H5 and H7 su... [Objective] The research aimed to design primers that are suitable for detecting H5 and H7 subtypes of avian influenza virus （AIV） ; [Method] DNAStar was used to analyze the homology of the sequences of H5 and H7 subtypes of AIV accessed in GenBank, and design primers（ by Primer Premier 5.0） on high homologous region of these sequences, and then amplified by RT-PCR. [Result] The multiplex RT-PCR amplification, agarose gel electrophoresis and sequencing results showed that the self-designed primers are successful for detecting AIV. [Conclusion] It is feasible to rapidly diagnose AIV through this method. 展开更多
关键词 Avian influenza virus Primer Premier 5.0 DNAStar Multiplex RT-PCR amplification
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Detection and Molecular Characterization of Enteroviruses in Korean Surface Water by Using Integrated Cell Culture Multiplex RT-PCR 认领 引用 被引量：1
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作者 GYUCHEOL LEE CHANHEE LEE +1 位作者 CHANSEUNG PARK SANGGI JEONG 《Biomedical and Environmental Sciences》 SCIE CAS 2008年第5期425-431,共7页
Objective To identify waterborne enteric viruses in Korean surface water. Methods Integrated cell culture（ICC）multiplex reverse transcription-polymerase chain reaction （RT-PCR） was simultaneously designed to detec... Objective To identify waterborne enteric viruses in Korean surface water. Methods Integrated cell culture（ICC）multiplex reverse transcription-polymerase chain reaction （RT-PCR） was simultaneously designed to detect coxsackieviruses （CV）, polioviruses （PV）, and reoviruses （RV）. ICC-multiplex RT-PCR and phylogenetic analysis were conducted using 21 total culturable virus assay （TCVA）-positive sample-inoculated cell cultures. Results CV and RV were detected in 9 samples each, and 3 samples were positive for both CV and RV. PV was not detected in any sample. Molecular phylogenetic analysis of the VP1 gene sequences revealed that CV types B2 and B4 predominated in Korean surface water, and the nucleotide sequences of CV type B2 were clustered with those of CVs isolated from China and Japan. The results suggested that the evolution of these viruses occurred in a region-specific manner. Conclusion CV and RV are detectable in Korean surface water, with a predominance of CV type B2, and the evolution of CV type B2 occur in a region-specific manner. 展开更多
关键词 ICC-multiplex RT-PCR Coxsackieviruses Molecular phylogeny
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Use of a Multiplex RT-PCR Assay for Simultaneous Detection of the North American Genotype Porcine Reproductive and Respiratory Syndrome Virus,Swine Influenza Virus and Japanese Encephalitis Virus 认领 引用 被引量：29
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作者 CHEN Hong-ying WEI Zhan-yong +6 位作者 ZHANG Hong-ying LüXiao-li ZHENG Lan-lan CUI Bao-an LIU Jinpeng ZHU Qian-lei WANG Zi-xin 《Agricultural Sciences in China》 2010年第7期1050-1057,共8页
A multiplex reverse transcriptase-polymerase chain reaction(multiplex RT-PCR) assay was developed and subsequently evaluated for its efficacy in the detection of multiple viral infections simultaneously,in swine.Speci... A multiplex reverse transcriptase-polymerase chain reaction(multiplex RT-PCR) assay was developed and subsequently evaluated for its efficacy in the detection of multiple viral infections simultaneously,in swine.Specific primers for each of the 3 RNA viruses,North American genotype porcine reproductive and respiratory syndrome virus,Japanese encephalitis virus,and swine influenza virus,were used in the testing procedure.The assay was shown to be highly sensitive because it could detect as little as 10-5 ng of each of the respective amplicons in a single sample containing a composite of all 3 viruses.The assay was also effective in detecting one or more of the same viruses in various combinations in specimens,including lymph nodes,lungs,spleens,and tonsils,collected from clinically ill pigs and in spleen specimens collected from aborted pig fetuses.The results from the multiplex RT-PCR were confirmed by virus isolation.The relative efficiency(compared to the efficiency of separate assays for each virus) and apparent sensitivity of the multiplex RT-PCR method show that this method has potential for application in routine molecular diagnostic procedures. 展开更多
关键词 Japanese encephalitis virus multiplex RT-PCR porcine reproductive and respiratory syndrome virus swine influenza virus
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Respiratory Virus Multiplex RT-PCR Assay Sensitivities and Influence Factors in Hospitalized Children with Lower Respiratory Tract Infections 认领 引用 被引量：16
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作者 Jikui Deng Zhuoya Ma +5 位作者 Wenbo Huang Chengrong Li Heping Wang Yuejie Zheng Rong Zhou Yi-Wei Tang 《Virologica Sinica》 CAS CSCD 2013年第2期97-102,共6页
Multiplex RT-PCR assays have been widely used tools for detection and differentiation of a panel of respiratory viral pathogens. In this study, we evaluated the Qiagen ResPlex lI V2.0 kit and explored factors influenc... Multiplex RT-PCR assays have been widely used tools for detection and differentiation of a panel of respiratory viral pathogens. In this study, we evaluated the Qiagen ResPlex lI V2.0 kit and explored factors influencing its sensitivity. Nasopharyngeal swab （NPS） specimens were prospectively collected from pediatric inpatients with lower respiratory tract infections at the time of admission in the Shenzhen Children＇s Hospital from May 2009 to April 2010. Total nucleic acids were extracted using the EZ1 system （Qiagen, Germany） and 17 respiratory viruses and genotypes including influenza A virus （FluA）, FluB, parainfluenza virus 1 （PIV1）, PIV2, PIV3, PIV4, respiratory syncytial virus （RSV）, human metapneumovirus （hMPV）, rhinoviruses （RhV）, enteroviruses （EnV）, human bocaviruses （hBoV）, adenoviruses （AdV）, four coronaviruses （229E, OC43, NL63 and HKU1）, and FluA 2009 pandemic H1NI（H1NI-p） were detected and identified by the ResPlex II kit. In parallel, 16 real-time TaqMan quantitative RT-PCR assays were used to quantitatively detect each virus except for RhV. Influenza and parainfluenza viral cultures were also performed. Among the total 438 NPS specimens collected during the study period, one or more viral pathogens were detected in 274 （62.6%） and 201（45.9%） specimens by monoplex TaqMan RT-PCR and multiplex ResPlex, respectively. When results from monoplex PCR or cell culture were used as the reference standard, the multiplex PCR possessed specificities of 92.9-100.0%. The sensitivity of multiplex PCR for PIV3, hMPV, PIV1 and BoV were 73.1%, 70%, 66.7% and 55.6%, respectively, while low sensitivities （11.1%-40.0%） were observed for FluA, EnV, OC43, RSV and H1N1. Among the seven viruses/genotypes detected with higher frequencies, multiplex PCR sensitivities were correlated significantly with viral loads determined by the TaqMan RT-PCR in FluA, H 1N 1-p and RSV （p=0.011-0.000） The Qiagen ResPlex II multiplex RT-PCR kit possesses excellent specificity for simultaneous detection of 17 viral pathogens in NPS specimens in pediatric inpatients at the time of admission. The sensitivity of multiplex RT-PCR was influenced by viral loads, specimen process methods, primer and probe design and amplification condition. 展开更多
关键词 Multiplex RT-PCR Respiratory viral loads Cell culture Lower respiratory tract infection
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Rapid Detection Co-infections of Classical Swine Fever Virus and Porcine Reproductive and Respiratory Syndrome Virus by One-step Multiplex RT-PCR 认领 引用 被引量：1
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作者 TIAN Hong WU Jinyan YAN Chen SHANG Youjun YIN Shuanghui LIU Xiangtao 《Journal of Northeast Agricultural University(English Edition)》 CAS 2011年第4期50-54,共5页
Classical swine fever virus （CSFV） and porcine reproductive and respiratory syndrome virus （PRRSV） have caused immense economic loss in the pig industry and are considered to be the two most important infectious d... Classical swine fever virus （CSFV） and porcine reproductive and respiratory syndrome virus （PRRSV） have caused immense economic loss in the pig industry and are considered to be the two most important infectious diseases of pigs in the world A multiplex reverse transcription polymerase chain reaction （multiplex RT-PCR） was developed for CSFV and PRRSV co-infections or infections, respectively. A set of two pairs of primer was designed based on the sequence of nonstructural protein NS54B of CSFV and ORF7 gene of PRRSV. The diagnostic accuracy of multiplex RT-PCR assay was evaluated by using 56 field clinical samples by multiplex RT-PCR, single RT-PCR and sequence analysis; and the specificity of multiplex PCR was verified by using constructed plasmids containing the specific viral target fragments of PRRSV and CSFV, respectively. The results indicated that this assay could reliably differentiate PRRSV and CSFV in co-infection samples. The multiplex RT-PCR developed in this study might provide a new avenue to the rapid the detection of CSFV and PRRSV in one reaction. 展开更多
关键词 CSFV PRRSV multiplex RT-PCR co-infection
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Detection of enteroviruses and hepatitis a virus in water by consensus primer multiplex RT-PCR 认领 引用 被引量：4
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作者 Jun-Wen Li Xin-Wei Wang Chang-Qing Yuan Jin-Lai Zheng Min Jin Nong Song Xiu-Quan Shi Fu-Huan Chao Department of Environment and Health,Institute of Health and Environmental Medicine of Tianiin,1 Da Li Road.Tianjin 300050,China 《World Journal of Gastroenterology》 SCIE CAS 2002年第4期699-702,共4页
AIM: To develop a rapid detection method ofenteroviruses and Hepatitis A virus (HAV).METHODS: A one-step, single-tube consensus primersmultiplex RT-PCR was developed to simultaneouslydetect Poliovirus, Coxsackie virus... AIM: To develop a rapid detection method ofenteroviruses and Hepatitis A virus (HAV).METHODS: A one-step, single-tube consensus primersmultiplex RT-PCR was developed to simultaneouslydetect Poliovirus, Coxsackie virus, Echovirus and HAV.A general upstream primer and a HAV primer and fourdifferent sets of primers (5 primers) specific forPoliovirus, Coxsacki evirus, Echovirus and HAV cDNAwere mixed in the PCR mixture to reverse transcriptand amplify the target DNA.Four distinct amplified DNAsegments representing Poliovirus, Coxsackie virus,Echovirus and HAV were identified by gelelectrophoresis as 589-,671-, 1084-, and 1128bpsequences, respectively. Semi-nested PCR was used toconfirm the amplified products for each enterovirus andHAV.RESULTS: All four kinds of viral genome RNA weredetected, and producing four bands which could bedifferentiated by the band size on the gel.To confirmthe specificity of the multiplex PCR products, semi-nested PCR was performed. For all the four strainstested gave positive results .The detection sensitivityof multiplex PCR was similar to that of monoplex RT-PCR which was 24 PFU for Poliovrus, 21 PFU forCoxsackie virus,60 PFU for Echovirus and 105 TCID50for HAV. The minimum amount of enteric viral RNAdetected by semi-nested PCR was equivalent to 2.4 PFUfor Poliovrus, 2.1 PFU for Coxsackie virus, 6.0 PFU forEchovirus and 10.5 TCID50 for HAV.CONCLUSION: The consensus primers multiplex RT-PCRhas more advantages over monoplex RT-PCR for entericviruses detection, namely, the rapid turnaround timeand cost effectiveness. 展开更多
关键词 肝炎病毒 肠病毒 RT-PCR 病毒检测
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Multiplex RT-PCR-based detections of CEA, CK20 and EGFR in colorectal cancer patients 认领 引用 被引量：22
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作者 Aikaterini Tsouma Chrysanthi Aggeli +7 位作者 Panagiotis Lembessis George N Zografos Dimitris P Korkolis Dimitrios Pectasides Maria Skondra Nikolaos Pissimissis Anastasia Tzonou Michael Koutsilieris 《World Journal of Gastroenterology》 SCIE CAS 2010年第47期5965-5974,共10页
AIM: To develop a multiplex reverse transcription polymerase chain reaction (RT-PCR) method detecting cir-culating tumor cells in the peripheral blood of colorectal cancer (CRC) patients. METHODS: Peripheral blood sam... AIM: To develop a multiplex reverse transcription polymerase chain reaction (RT-PCR) method detecting cir-culating tumor cells in the peripheral blood of colorectal cancer (CRC) patients. METHODS: Peripheral blood samples were collected from 88 CRC patients and 40 healthy individuals from the blood donors' clinic and subsequently analyzed by multiplex RT-RCR for the expression of carcinoembryonic antigen (CEA), cytokeratin 20 (CK20) and epidermal growth factor receptor (EGFR) mRNA. The analysis involved determining the detection rates of CEA, CK20 and EGFR transcripts vs disease stage and overall survival. Median follow-up period was 19 mo (range 8-28 mo). RESULTS: Rates of CEA, CK20 and EGFR detection in CRC patients were 95.5%, 78.4% and 19.3%, respectively. CEA transcripts were detected in 3 healthy volunteer samples (7.5%), whereas all control samples were tested negative for CK20 and EGFR transcripts. The increasing number of positive detections for CEA, CK20 and EGFR transcripts in each blood sample was positively correlated with Astler-Coller disease stage (P< 0.001) and preoperative serum levels of CEA (P=0.029) in CRC patients. Data analysis using Kaplan-Meier estimator documented signif icant differences in the overall survival of the different CRC patient groups as formed according to the increasing number of positivity for CEA, CK20 and EGFR transcripts. CONCLUSION: These data suggest that multiplex RTPCR assay can provide useful information concerning disease stage and overall survival of CRC patients. 展开更多
关键词 Peripheral blood Carcinoembryonic antigen Cytokeratin 20 Epidermal growth factor receptor Multiplex reverse transcription polymerase chain reaction
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Simultaneous detection of duck hepatitis A virus types 1 and 3, and of duck astrovirus type 1, by multiplex RT-PCR 认领 引用 被引量：3
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作者 Linlin Chen Mingjie Ma +5 位作者 Ruihua Zhang Qian Xu Xingkui Si Yu Wang Zhijing Xie Shijin Jiang 《Virologica Sinica》 CAS CSCD 2014年第3期196-198,共3页
Dear Editor,Duck virus hepatitis(DVH)is caused by at least threedifferent RNA viruses,including duck hepatitis A virus(DHAV),duck astrovirus type 1(DAstV-1),and duckastrovirus type 2(DAstV-2).The first of these,DHAV,h... Dear Editor,Duck virus hepatitis(DVH)is caused by at least threedifferent RNA viruses,including duck hepatitis A virus(DHAV),duck astrovirus type 1(DAstV-1),and duckastrovirus type 2(DAstV-2).The first of these,DHAV,has been classified into three serotypes by 展开更多
关键词 鸭肝炎病毒 病毒类型 多重RT-PCR 同时检测 鸭病毒性肝炎 RNA病毒 DVH 2型
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Detection of the Mex Efflux Pumps in Pseudomonasaeruginosaby Using a Combined Resistance-Phenotypic Markers and Multiplex RT-PCR 认领 引用
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作者 Kanchana Poonsuk Rungtip Chuanchuen 《Open Journal of Medical Microbiology》 2014年第3期153-160,共8页
The aim of this study was to detect the expression of 4 clinically-important efflux pumps in the Resistance-Nodulation-Cell Division (RND) family including MexAB-OprM, MexXY, MexCD-OprJ and MexEF-OprN in Pseudomonas a... The aim of this study was to detect the expression of 4 clinically-important efflux pumps in the Resistance-Nodulation-Cell Division (RND) family including MexAB-OprM, MexXY, MexCD-OprJ and MexEF-OprN in Pseudomonas aeruginosa using a combination of resistance-phenotypic markers and multiplex RT-PCR (mRT-PCR). The antibiotic substrates specific for each Mex systems were used as phenotypic markers including carbenicillin, MexAB-OprM, erythromycin, MexCD-OprJ, norfloxacin and imipenem, MexEF-OprN and gentamicin, MexXY-OprM. The methods were validated with reference strains with known genotypes of the Mex systems and the potential applicability in clinical practice was tested with clinical isolates. The results for the reference strains support that the combination of resistance phenotype and mRT-PCR is a potential-attractive method for diagnosis of efflux-mediated resistance in P. aeruginosa. Further development to make it more practical for clinical use and study in a larger number of clinical isolates is required. 展开更多
关键词 Multidrug Efflux Pumps Multiplex RT-PCR Pseudomonas aeruginosa Resistance-Phenotypic Marker
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Establishment of Multiplex RT-PCR Detection System for Three Viruses in Freesia 认领 引用
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作者 Fan Ronghui Huang Minling +1 位作者 Wu Jianshe Luo Yuanhua 《Plant Diseases and Pests》 2013年第1期33-35,38,共3页
The specific primers were designed according to conserved sequences of coat protein （CP） gene of Freesia mosaic virus （FreMV）, Cucumber mosaic vi- rus （CMV） and Bean yellow mosaic virus （BYMV） published in Gen... The specific primers were designed according to conserved sequences of coat protein （CP） gene of Freesia mosaic virus （FreMV）, Cucumber mosaic vi- rus （CMV） and Bean yellow mosaic virus （BYMV） published in GenBank, and a multiplex PCR protocol for simultaneous detection of these three viruses in freesia was developed. Three specific fragments were simultaneously amplified in a single PCR reaction. Their lengths were determined to be 340,628 and 212 bp, respec-tively. The sequence analysis indicated that three viruses shared at least 97% of homology with reference sequence. Sensitivity test showed that these three viruses could be detected out in the infected plant tissue greater than 10^-2 mg. 展开更多
关键词 Freesia Virus Freesia mosaic virus Cucumber mosaic virus Bean yellow mosaic virus Multiplex PCR
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鸭蛋减产综合征病毒巢式RT-PCR方法的建立及应用 认领 引用 被引量：1
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作者 张志飞 宿鑫 +10 位作者 硕盾 罗燕秋 闫大为 潘学 黄东明 温向阳 许榜丰 滕巧泱 刘芹防 苑纯秀 李泽君 《中国动物传染病学报》 CAS 北大核心 2026年第1期114-118,共5页
本研究基于鸭蛋减产综合征病毒基因序列设计两对检测引物,通过反应体系和条件的优化,建立了DERSV巢式RT-PCR检测方法。通过特异性、灵敏性、重复性以及临床样品检测验证方法的实用性和可靠性。试验结果显示,建立的套式RT-PCR方法灵敏性... 本研究基于鸭蛋减产综合征病毒基因序列设计两对检测引物,通过反应体系和条件的优化,建立了DERSV巢式RT-PCR检测方法。通过特异性、灵敏性、重复性以及临床样品检测验证方法的实用性和可靠性。试验结果显示,建立的套式RT-PCR方法灵敏性比普通RT-PCR方法高1000倍,特异性试验结果显示该方法仅能从DERSV扩增出预期目的条带,其他7种常见禽病均为阴性。综上,本研究成功建立一种灵敏性高、特异性强的DERSV巢式RT-PCR检测方法,为临床快速准确诊断鸭蛋减产综合征提供了技术手段。 展开更多
关键词 鸭蛋减产综合征病毒 巢式RT-PCR 检测
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Silicon photonic microring-based eight-channel wavelength-division multiplexing transceiver for high-density optical interconnects 认领 引用 被引量：1
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作者 Shenlei Bao Chao Cheng +6 位作者 Xianglin Bu Houyou Lai Xishan Yu Jianjun Zhou Jintao Xue Wenfu Zhang Binhao Wang 《Advanced Photonics Nexus》 CSCD 2026年第3期82-96,共15页
We demonstrate a fully integrated eight-channel dense wavelength-division multiplexing silicon photonic transceiver supporting 200-Gbps per-channel PAM4 operation,enabling a total chip-to-chip data rate of 1.6 Tbps.Th... We demonstrate a fully integrated eight-channel dense wavelength-division multiplexing silicon photonic transceiver supporting 200-Gbps per-channel PAM4 operation,enabling a total chip-to-chip data rate of 1.6 Tbps.The transmitter employs compact single-bus microring modulators,whereas the receiver adopts a polarization diversity architecture based on cascaded dual-ring filters and integrates a bidirectionally incident photodetector,maintaining stable performance under arbitrary input polarization.A unified multichannel thermo-optic feedback architecture is implemented at both the transmitter and receiver,enabling cooperative link-level wavelength alignment without pre-calibration.This multi-channel parallel control scheme reduces wavelength locking time by~30×while achieving fine wavelength-tracking accuracy of 2.74 pm with negligible thermal overhead.Comprehensive device-and system-level experiments validate the robustness and scalability of the proposed architecture.We uniquely address the critical bottlenecks of high polarization sensitivity and latency in wavelength alignment through a highly integrated silicon photonic architecture.By implementing polarization-splitting grating couplers and synchronized wavelengthlocking schemes,we provide a transformative solution for high-density co-packaged optics.Our approach significantly reduces system footprint,enhances operational reliability,and improves power efficiency,thereby bridging the gap between laboratory demonstrations and practical 1.6-Tbps scale chip-to-chip interconnects. 展开更多
关键词 silicon photonics optical interconnects co-packaged optics optical input/output micro-ring modulators silicongermanium photodiodes wavelength-division multiplexing wavelength stabilization thermal control
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五种羊病毒多重实时荧光RT-PCR检测方法的建立与应用 认领 引用
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作者 王素秋 万本屹 +7 位作者 赵相鹏 张伟 白子龙 杜思乐 吴丹 张小寒 史喜菊 张灿 《中国兽医科学》 CAS CSCD 北大核心 2026年第8期1077-1085,共9页
为建立一种可同步检测和鉴别蓝舌病病毒(BTV)、口蹄疫病毒(FMDV)、施马伦贝格病毒(SBV)、小反刍兽疫病毒(PPRV)、绵羊痘和山羊痘病毒(SPPV-GTPV)的方法,针对上述病毒的保守基因序列设计特异性引物和探针,通过对反应条件多次优化,建立了... 为建立一种可同步检测和鉴别蓝舌病病毒(BTV)、口蹄疫病毒(FMDV)、施马伦贝格病毒(SBV)、小反刍兽疫病毒(PPRV)、绵羊痘和山羊痘病毒(SPPV-GTPV)的方法,针对上述病毒的保守基因序列设计特异性引物和探针,通过对反应条件多次优化,建立了上述5种病毒的多重实时荧光RT-PCR检测方法,并与相应的单重实时荧光RT-PCR方法进行比较。标准曲线分析显示,多重与单重方法的扩增效率(E值)均在90%~100%之间,相关系数(R2)均≥0.996,多重反应体系未产生明显的相互干扰或灵敏度降低现象。多重荧光RT-PCR检测方法对各靶标的最低检测限为5.31 copies/μL,特异性为100%。对6份BTV、4份PPRV和10份FMDV阳性样品检测的符合率为100%。对来自中国西北地区的500份临床样品进行检测,共检出PPRV阳性24份、FMDV阳性10份和BTV阳性1份,检测结果与单重检测方法完全一致。研究结果表明,本研究中建立的多重实时荧光RT-PCR方法可实现BTV、FMDV、SBV、PPRV和SPPV-GTPV的同步检测与鉴别,适用于临床上混合感染的快速筛查。 展开更多
关键词 多重实时荧光RT-PCR 蓝舌病病毒 口蹄疫病毒 施马伦贝格病毒 小反刍兽疫病毒 绵羊痘和山羊痘病毒 混合感染
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内蒙古自治区葡萄病毒RT-PCR检测及序列分析 认领 引用 被引量：1
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作者 李正男 张璐 +4 位作者 闫雨婷 郭燕玲 陈晓旭 孙婧 张磊 《山西农业大学学报（自然科学版）》 CAS 北大核心 2026年第2期116-127,共12页
[目的]明确内蒙古自治区葡萄产区病毒种类、检出率、发生特点和分类地位。[方法]2023年6月至10月,从呼和浩特市、包头市、鄂尔多斯市、巴彦淖尔市、乌海市、赤峰市以及通辽市采集69份葡萄样品,提取总RNA后通过RT-PCR进行带毒情况检测,... [目的]明确内蒙古自治区葡萄产区病毒种类、检出率、发生特点和分类地位。[方法]2023年6月至10月,从呼和浩特市、包头市、鄂尔多斯市、巴彦淖尔市、乌海市、赤峰市以及通辽市采集69份葡萄样品,提取总RNA后通过RT-PCR进行带毒情况检测,检测为阳性的样品对目标片段进行回收、克隆和测序。对测序结果通过生物信息学软件进行核酸一致性分析和系统发育分析。[结果]检测的病毒共14种,其中9种在内蒙古发生,检出率从高到低分别为GFkV(68.10%)、GRSPaV(63.80%)、GLRaV-3(39.10%)、GPGV(23.20%)、GFabV(8.70%)、GINV(7.20%)、GVE(5.80%)、GLRaV-2(2.90%)和GVB(2.90%)。同一样品复合感染(≥2种病毒)现象普遍,在43个样品(62.3%)中发现复合侵染。GFkV核酸序列一致性分析结果显示,GFkV 29-2与GFkV 8-2的一致性最低为81.3%,系统发育树表明,GFkV 29-2与墨西哥的GFkV Vitis-TrPADL45(PQ473685)亲缘关系较近。GRSPaV核酸序列一致性分析结果显示,GRSPaV 31-1和GRSPaV 8-T的一致性最低为78.4%;GRSPaV 32-1和GRSPaV8-T的一致性最低为78.4%。系统发育树表明,内蒙古不同地区发现的GRSPaV和不同国家的GRSPaV聚集在一起,形成亲缘关系较近的分支,GRSPaV 31-1、GRSPaV 32-1与南非17-8548(MK077682)亲缘关系较近。[结论]本研究首次明确了内蒙古自治区葡萄产区病毒种类、检出率、发生特点及检出病毒的分类地位。研究结果为葡萄病毒病的绿色防控提供了数据基础,开发葡萄无病毒苗木是上述问题的有效解决手段。 展开更多
关键词 内蒙古自治区 葡萄病毒 RT-PCR 系统发育分析
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Digital microfluidics actuated multiplexed heterogeneous immunoassay with enhanced automation and efficiency 认领 引用
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作者 Mengyuan Zhao Xianming Liu +10 位作者 Haiyue Ye Dachuan Ma Yue Hou Hongyu Dong Yangyang Long Ningning Liu Xiaoxu Wang Xiaodan Yu Yahong Chen Xinmiao Liang Yao Lu 《Chinese Chemical Letters》 SCIE CAS CSCD 2026年第4期688-693,共6页
Digital microfluidics(DMF)shows great promise in addressing the need for miniaturization and automation in immunoassay detection.Despite recent advances,an automatically operated,multiplexed heterogeneous immunoassay ... Digital microfluidics(DMF)shows great promise in addressing the need for miniaturization and automation in immunoassay detection.Despite recent advances,an automatically operated,multiplexed heterogeneous immunoassay platform powered by DMF remains underdeveloped.Here we present a DMF platform for automated and multiplexed heterogeneous immunoassay detection by coupling spatial barcoding with automatic and uniform droplet dispensing.FluoroPel was selected as a robust hydrophobic reagent for coating the DMF top plate,and it also served as the substrate for the immuno-reaction.Its mechanical robustness was further enhanced with a Cytop CTL-809A adhesive layer under the top hydrophobic layer.Hourglass-shaped electrode patterns ensured consistent and uniform distribution of immunoassay reagents,with volume variation down to 1.0%.The analysis duration was significantly reduced from 75 min to 20 min after a heating module was integrated to elevate the immuno-reaction temperature to 37℃.Utilizing a compact instrument featuring a multi-droplet manipulation protocol,we successfully implemented fully operated,multi-sample,multiplexed immunoassays using recombinant proteins on cell culture supernatants on the DMF platform.This innovative platform significantly enhances the efficiency,reliability,and degree of automation of DMF-actuated multiplexed heterogeneous immunoassays,potentially providing a viable solution for field deployment and multi-sample parallel diagnosis. 展开更多
关键词 Digital microfluidics Mulitpled immunoassay Automation Multiplexed FluoroPel
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Ambient-energy-driven space-time-coding metasurface for spacefrequency-division multiplexing wireless communications 认领 引用
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作者 Han Wei Tian Chao Song +3 位作者 Dong Jie Wang Qian Zhu Tie Jun Cui Wei Xiang Jiang 《Opto-Electronic Advances》 SCIE EI CAS CSCD 2026年第2期12-26,共15页
Programmable metasurfaces have shown exceptional potentials in wireless communications due to their capability to manipulate electromagnetic(EM)waves dynamically and flexibly.However,the large-scale application and fl... Programmable metasurfaces have shown exceptional potentials in wireless communications due to their capability to manipulate electromagnetic(EM)waves dynamically and flexibly.However,the large-scale application and flexible deployment of programmable metasurfaces still face challenges of high communication capacity requirements and stringent energy constraints.Here,we report an ambient-energy-driven space-time-coding metasurface to address these issues.On one hand,the metasurface can achieve efficient space-frequency-division multiplexing manipulations by dynamically controlling multiple frequencies and the spatial propagation directions of reflection EM waves.On the other hand,the shared-aperture ambient solar energy harvesting capability and low power consumption characteristic of the metasurface enable it to be self-powered without relying on any external power supply.To demonstrate these remarkable features,a four-channel wireless communication system prototype is built using the programmable metasurface.Experimental results confirm that four distinct images can be transmitted to four user terminals simultaneously,independently,and in real time with remarkably low energy consumption per bit.Such innovative metasurface provides a simple and effective approach for integrating ambient energy harvesting,multidimensional microwave manipulation,and direct information modulation on a single physical platform,which will advance the wireless communications in cost-effectiveness,enhanced capacity,energy efficiency,and environmental friendliness. 展开更多
关键词 programmable metasurfaces space-time-coding metasurfaces ambient energy harvesting self-powered multiplexing wireless communications energy-efficient
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日本黄瓜绿斑驳花叶病毒常规和实时荧光RT-PCR检测 认领 引用
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作者 陈红运 叶明辉 +3 位作者 陈青 廖富荣 于子翔 沈建国 《植物检疫》 CAS 2026年第1期53-57,共5页
本研究以日本黄瓜绿斑驳花叶病毒(kyuri green mottle mosaic virus,KGMMV)为材料,设计1对常规RT-PCR检测引物KGCPN-F/KGCPN-R和1组引物/探针KGM-F/KGM-R/KGM-P,分析测试了2对已发表KGMMV常规RT-PCR检测引物KGCP-F/KGCP-R和KGMP-F/KGMP... 本研究以日本黄瓜绿斑驳花叶病毒(kyuri green mottle mosaic virus,KGMMV)为材料,设计1对常规RT-PCR检测引物KGCPN-F/KGCPN-R和1组引物/探针KGM-F/KGM-R/KGM-P,分析测试了2对已发表KGMMV常规RT-PCR检测引物KGCP-F/KGCP-R和KGMP-F/KGMP-R,建立了KGMMV的常规和实时荧光RT-PCR检测方法。结果表明,引物KGCP-F/KGCP-R扩增KGMMV时出现预期大小的条带,扩增黄瓜绿斑驳花叶病毒(cucumber green mottle mosaic virus,CGMMV)时出现非常微弱的条带;引物KGCPN-F/KGCPN-R扩增KGMMV时出现预期大小的条带,扩增小西葫芦绿斑驳花叶病毒(zucchini green mottle mosaic virus,ZGMMV)时出现比预期稍大的条带,通过对PCR产物进行序列测定和分析比对可准确鉴定KGMMV。KGCP-F/KGCP-R和KGCPN-F/KGCPN-R的相对灵敏度分别为10-6和10-5稀释度,适用于KGMMV的常规RT-PCR检测。基于引物探针KGM-F/KGM-R/KGM-P建立的KGMMV实时荧光RT-PCR检测方法能特异性检出KGMMV,相对灵敏度达10-7稀释度,分别比2对常规RT-PCR检测引物高10倍和100倍,适用于瓜类种子中KGMMV的快速检测。 展开更多
关键词 日本黄瓜绿斑驳花叶病毒 常规RT-PCR 实时荧光RT-PCR 检测
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Terabit nonlinear frequency division multiplexing transmission desensitized to laser frequency offset and phase noise 认领 引用
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作者 Xiang Cai Xiansong Fang +4 位作者 Jiayu Zheng Yixiao Zhu Xinyu Chen Xian Zhou Fan Zhang 《Advanced Photonics Nexus》 CSCD 2026年第2期90-99,共10页
Nonlinear frequency division multiplexing(NFDM)utilizes the nonlinear Fourier transform(NFT)to decompose signals into the nonlinear frequency domain that inherently resists fiber Kerr nonlinearity distortions.However,... Nonlinear frequency division multiplexing(NFDM)utilizes the nonlinear Fourier transform(NFT)to decompose signals into the nonlinear frequency domain that inherently resists fiber Kerr nonlinearity distortions.However,practical NFDM has been hindered by sensitivity to laser frequency offset and phase noise.In conventional NFDM receivers,a residual frequency offset typically remains before the NFT,whereas phase recovery performed after the NFT can only estimate the average phase rotation per burst,fundamentally limiting tolerance to frequency-phase impairments.In this work,we propose and experimentally demonstrate a residual carrier scheme to compensate for frequency offset and phase noise before the receiver NFT with sampling-point-level precision.Using a 3 MHz distributed feedback(DFB)laser,this scheme enables 1.0 Tb∕s PS-256 QAM NFDM transmission over 320 km,with a high spectral efficiency of 9.85 b∕s∕Hz.The Q2-factor degradation is only 0.85 dB as the total linewidth increases from 200 Hz to 3.1 MHz.The scheme achieves a Q2-factor improvement of 2.6 dB with 100 k Hz external cavity lasers and>6 dB with a 3 MHz DFB laser,compared with the conventional time-domain pilot method.To our knowledge,this is the first time NFDM demonstrates comparable tolerance to frequency-phase impairments as single-carrier systems.The proposed scheme desensitizes NFDM to frequency-phase impairments,overcoming a major barrier to practical deployment. 展开更多
关键词 nonlinear frequency division multiplexing nonlinear Fourier transform frequency offset phase noise residual carrier
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苹果软枝病毒1巢式RT-PCR检测方法的建立及应用 认领 引用
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作者 刘成龙 范旭东 +2 位作者 任芳 胡国君 董雅凤 《植物病理学报》 CAS CSCD 北大核心 2026年第1期181-184,共4页
苹果软枝病(apple rubbery wood disease, ARWD)是一种重要的苹果病毒病害,最早于1935年在英国苹果‘Lord Lambourne’上被发现[1-2]。主要影响木质素合成,进而使苹果的茎和枝条异常柔软,容易弯曲折断;还可以引起节间缩短,发育延迟,使... 苹果软枝病(apple rubbery wood disease, ARWD)是一种重要的苹果病毒病害,最早于1935年在英国苹果‘Lord Lambourne’上被发现[1-2]。主要影响木质素合成,进而使苹果的茎和枝条异常柔软,容易弯曲折断;还可以引起节间缩短,发育延迟,使树体对冷胁迫的敏感性增加[3-5]。在种植‘金冠’、‘嘎啦’等感病品种的地区或国家,可导致苹果减产30%[6-7]。 展开更多
关键词 木质素合成 苹果软枝病 RT-PCR检测
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