A multiplex polymerase chain reaction （PCR） was developed to detect three pathogenic genes of enteropathogenic, enterotocigenic and enteroinvasive Escherichia coli In this study three different sets of oligonucleoti...A multiplex polymerase chain reaction （PCR） was developed to detect three pathogenic genes of enteropathogenic, enterotocigenic and enteroinvasive Escherichia coli In this study three different sets of oligonucleotide primer were simultaneously used, and in this way, specific fragments of 880, 600, 150 bp for EPEC eaeA, EIEC ipaH and ETEC ST genes were amplified, respectively. The best condition of the multiplex PCR was： after an initial heat denaturation step at 95℃for 5 min, followed by 30 cycles of denaturation at 94 ℃ for 40 s, primer annealing at 51.3℃ for 40 s and extension at 72 ℃ for 1 min, final extension at 72 ℃ for 10 min. The detection limit of the eaeA, ipaH and ST primers was 38.7423, 3.60519, 29.9448 ng·mL^-1 （4.3×10^4, 1.5×10^3, 2.6×10^4 CFU·mL^-1）, respectively. It may be a good way for the detection and identification of Diarrhea-causing E. coli.展开更多
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
Potato variety Kexin18 was used as testing materials in this research to study the influence on main components in multiplex PCR system, different primer ratios and annealing temperatures in SSR marker amplification. ...Potato variety Kexin18 was used as testing materials in this research to study the influence on main components in multiplex PCR system, different primer ratios and annealing temperatures in SSR marker amplification. Concentration and gradient experiments for four components （enzyme, MgCl2, DNA template and dNTPs） in PCR system were used in the research with the concentration of the other component remained the same; the orthogonal design L9 （34） was applied in the optimization of four sets of primers （STM0014, Pat, SSI, and UGP） in the reaction system at three levels; the temperature gradient selection was used to find out the optimum annealing temperature for the primer. The optimized multiplex PCR system of potato SSR marker with a total volume of 20 μL ： 2.5 μL 25 mmol.L-1 MgCl2, 0.6 μL 10 mmol·L-1 dNTPs, 0.8 U Taq, 80 ng DNA template was ultimately established through the comparison and analysis of test results; the ratio of four pairs of 4 mmol. L1 primers was 2 ： 1 ： 2 ： 3, and the annealing temperature was 54.7℃. The optimized reaction system could be repeated stably; and the stable and reliable amplification results were able to clearly distinguish different potato varieties. This research built the solid foundation for the further study of genetic diversity of potato germplasms and construction of DNA fingerprinting..展开更多
Smart antenna has been regarded as one promising technology to enhance the performance of CDMA mobile communication systems. However, when applied to 3G systems, the performance of traditional adaptive arrays may degr...Smart antenna has been regarded as one promising technology to enhance the performance of CDMA mobile communication systems. However, when applied to 3G systems, the performance of traditional adaptive arrays may degrade due to code and time multiplexing in dedicated physical channels. A novel semiblind adaptive array approach is proposed to solve this problem. It overcomes the selfinterfering problem by introducing a quasidespreading technique for the control channel, and contains a timemultipexed structure to utilize both pilot symbols and unknown control symbols within the control channel. The blind part of the proposed approach is based on the competition of two parallel branches with different reference sequences. Simulation results show significant improvement can be achieved by the proposed approach in a fastfading WCDMA environment.展开更多
In order to make systems that are based on unreliable components reliable, the design of fault tolerant architectures will be necessary. Inspired by von Neumann＇s negative AND（NAND）multiplexing and William＇s inter...In order to make systems that are based on unreliable components reliable, the design of fault tolerant architectures will be necessary. Inspired by von Neumann＇s negative AND（NAND）multiplexing and William＇s interwoven redundant logic, this paper presents a fault tolerant technique based on redundancy-modified NAND gates for future nanocomputers. Bifurcation theory is used to analyze fault tolerant ability of the system and the simulation results show that the new system has a much higher fault tolerant ability than the conventional multiplexing based on NAND gates.According to the evaluation, the proposed architecture can tolerate a device error rate of up to 10-1, with multiple redundant components. This fault tolerant technique is potentially useful for future nanoelectronics.展开更多
A cross-linkable fluorinated poly(ether ether ketone)(FPEEK)was synthesized for the fabrication of arrayed waveguide grating(AWG)multiplexer.The results of thermal gravimetric analysis(TGA)and near-infrared absorption...A cross-linkable fluorinated poly(ether ether ketone)(FPEEK)was synthesized for the fabrication of arrayed waveguide grating(AWG)multiplexer.The results of thermal gravimetric analysis(TGA)and near-infrared absorption spectrum show that the materials have high thermal stability and high optical transparency in the infrared communication region.The refractive index of FPEEK can be controlled easily by changing the fluorine content of the materials.The 32-channel AWG multiplexer is fabricated using the FPEEK and oxygen reactive ion etching technology.The AWG multiplexer exhibits that the insertion loss is from 12.8 to 17.8 dB and the channel crosstalk is less than-20 dB.The wavelength channel spacing and the center wavelength are 0.8nm and 1548nm,respectively.展开更多
A multiplex reverse transcription polymerase chain reaction (multiplex RT-PCR) was developed for the detection of porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV) and porcine ...A multiplex reverse transcription polymerase chain reaction (multiplex RT-PCR) was developed for the detection of porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV) and porcine group A rotavirus (GAR). Three pairs of primers were designed to target the M gene, N gene, and VP7 gene of PEDV, TGEV, GAR, respectively, and the multiplex RT-PCR was developed and optimized. The results of the multiplex RT-PCR and routine single RT-PCRs were compared using samples collected in the field. In laboratory testing, the detection limit of the multiplex RT-PCR is ～35 pg RNA of combined TGEV-PEDV-GAR vaccine. In the field trial, 75 fecal specimens collected from pigs with diarrhea, in the central area of China, were simultaneously tested by the multiplex RT-PCR and by routine single RT-PCRs to evaluate the relative sensitivity and specificity of the multiplex RT-PCR. The results indicate that this new assay is equal in quality to the routine RT-PCR assays (sensitivities were 92%, 100%, 100% for PEDV, TGEV, GAR, respectively; specificity was 100% for all three viruses). The multiplex RT-PCR, with high sensitivity and specificity, provides a new and alternative tool for the detection of PEDV, TGEV and GAR.展开更多
This review presents a reflection-type holographic memory using three-dimensional(3D)speckle-shift multiplexing. First, the schematic of the proposed memory system was described. Then,experimental demonstrations of mu...This review presents a reflection-type holographic memory using three-dimensional(3D)speckle-shift multiplexing. First, the schematic of the proposed memory system was described. Then,experimental demonstrations of multiplexing in plane and along the depth direction were presented. The estimated storage capacity of single layer recording was introduced and the maximum storage capacity was discussed. To increase the storage capacity, the multi-layered recording was described. In the multilayered recording, the storage capacity can be increased by appropriate arrangement of holograms in each layer.展开更多
Streptococcus suis is an important zoonotic agent causing severe diseases in both pigs and humans.To date,33 serotypes of S.suis have been identified based on antigenic differences in the capsular polysaccharide.The c...Streptococcus suis is an important zoonotic agent causing severe diseases in both pigs and humans.To date,33 serotypes of S.suis have been identified based on antigenic differences in the capsular polysaccharide.The capsular polysac-charide synthesis(cps)locus encodes proteins/enzymes that are responsible for capsular production and structure variation,which is the basis of S.suis serotyping.The cps clusters of 33 reference serotypes were published.Based on serotype-specific gene wzy in the cps gene cluster,we developed multiplex PCR(mPCR)assays for the known 33 serotypes.Seven new cps loci types were discovered,an mPCR assay for identifying them was developed in this study.The detection limit of the mPCR assay for the genomic DNA of representative strains of each new cps loci was 100 pg.Moreover,this mPCR strategy was applied in 91 untypable S.suis strains,and it turned out that 71 of them belong to these new cps types.The multiplex PCR assays deve-loped in this study provide a rapid and specific method for identification of 7 new cps types of S.suis.展开更多
基金Key Item of National Technology Research Project (2002BA518A06)Heilongjiang Province Department Fund (10541021)
文摘A multiplex polymerase chain reaction （PCR） was developed to detect three pathogenic genes of enteropathogenic, enterotocigenic and enteroinvasive Escherichia coli In this study three different sets of oligonucleotide primer were simultaneously used, and in this way, specific fragments of 880, 600, 150 bp for EPEC eaeA, EIEC ipaH and ETEC ST genes were amplified, respectively. The best condition of the multiplex PCR was： after an initial heat denaturation step at 95℃for 5 min, followed by 30 cycles of denaturation at 94 ℃ for 40 s, primer annealing at 51.3℃ for 40 s and extension at 72 ℃ for 1 min, final extension at 72 ℃ for 10 min. The detection limit of the eaeA, ipaH and ST primers was 38.7423, 3.60519, 29.9448 ng·mL^-1 （4.3×10^4, 1.5×10^3, 2.6×10^4 CFU·mL^-1）, respectively. It may be a good way for the detection and identification of Diarrhea-causing E. coli.
文摘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.
基金Supported by the International Cooperation Project of Heilongjiang Science and Technology Department (WC05B08)
文摘Potato variety Kexin18 was used as testing materials in this research to study the influence on main components in multiplex PCR system, different primer ratios and annealing temperatures in SSR marker amplification. Concentration and gradient experiments for four components （enzyme, MgCl2, DNA template and dNTPs） in PCR system were used in the research with the concentration of the other component remained the same; the orthogonal design L9 （34） was applied in the optimization of four sets of primers （STM0014, Pat, SSI, and UGP） in the reaction system at three levels; the temperature gradient selection was used to find out the optimum annealing temperature for the primer. The optimized multiplex PCR system of potato SSR marker with a total volume of 20 μL ： 2.5 μL 25 mmol.L-1 MgCl2, 0.6 μL 10 mmol·L-1 dNTPs, 0.8 U Taq, 80 ng DNA template was ultimately established through the comparison and analysis of test results; the ratio of four pairs of 4 mmol. L1 primers was 2 ： 1 ： 2 ： 3, and the annealing temperature was 54.7℃. The optimized reaction system could be repeated stably; and the stable and reliable amplification results were able to clearly distinguish different potato varieties. This research built the solid foundation for the further study of genetic diversity of potato germplasms and construction of DNA fingerprinting..
文摘Smart antenna has been regarded as one promising technology to enhance the performance of CDMA mobile communication systems. However, when applied to 3G systems, the performance of traditional adaptive arrays may degrade due to code and time multiplexing in dedicated physical channels. A novel semiblind adaptive array approach is proposed to solve this problem. It overcomes the selfinterfering problem by introducing a quasidespreading technique for the control channel, and contains a timemultipexed structure to utilize both pilot symbols and unknown control symbols within the control channel. The blind part of the proposed approach is based on the competition of two parallel branches with different reference sequences. Simulation results show significant improvement can be achieved by the proposed approach in a fastfading WCDMA environment.
基金supported by the National Natural Science Foundation of China(61571149)
文摘In order to make systems that are based on unreliable components reliable, the design of fault tolerant architectures will be necessary. Inspired by von Neumann＇s negative AND（NAND）multiplexing and William＇s interwoven redundant logic, this paper presents a fault tolerant technique based on redundancy-modified NAND gates for future nanocomputers. Bifurcation theory is used to analyze fault tolerant ability of the system and the simulation results show that the new system has a much higher fault tolerant ability than the conventional multiplexing based on NAND gates.According to the evaluation, the proposed architecture can tolerate a device error rate of up to 10-1, with multiple redundant components. This fault tolerant technique is potentially useful for future nanoelectronics.
基金Project(60223001,60177022)supported by the National Natural Science Foundation of Chinaproject(2001AA312160)supported by the National High Technology Research and Development Program of Chinaproject(TG2000036602)supported by the National Basic Research Program of China
文摘A cross-linkable fluorinated poly(ether ether ketone)(FPEEK)was synthesized for the fabrication of arrayed waveguide grating(AWG)multiplexer.The results of thermal gravimetric analysis(TGA)and near-infrared absorption spectrum show that the materials have high thermal stability and high optical transparency in the infrared communication region.The refractive index of FPEEK can be controlled easily by changing the fluorine content of the materials.The 32-channel AWG multiplexer is fabricated using the FPEEK and oxygen reactive ion etching technology.The AWG multiplexer exhibits that the insertion loss is from 12.8 to 17.8 dB and the channel crosstalk is less than-20 dB.The wavelength channel spacing and the center wavelength are 0.8nm and 1548nm,respectively.
基金supported by the National Key Research&Development Program of China(Grant No.2016YFD0600901)the National Natural Science Foundation of China(Grant No.31000294)the Priority Academic Program Development of Jiangsu Higher Education Institutions
文摘A multiplex reverse transcription polymerase chain reaction (multiplex RT-PCR) was developed for the detection of porcine epidemic diarrhea virus (PEDV), porcine transmissible gastroenteritis virus (TGEV) and porcine group A rotavirus (GAR). Three pairs of primers were designed to target the M gene, N gene, and VP7 gene of PEDV, TGEV, GAR, respectively, and the multiplex RT-PCR was developed and optimized. The results of the multiplex RT-PCR and routine single RT-PCRs were compared using samples collected in the field. In laboratory testing, the detection limit of the multiplex RT-PCR is ～35 pg RNA of combined TGEV-PEDV-GAR vaccine. In the field trial, 75 fecal specimens collected from pigs with diarrhea, in the central area of China, were simultaneously tested by the multiplex RT-PCR and by routine single RT-PCRs to evaluate the relative sensitivity and specificity of the multiplex RT-PCR. The results indicate that this new assay is equal in quality to the routine RT-PCR assays (sensitivities were 92%, 100%, 100% for PEDV, TGEV, GAR, respectively; specificity was 100% for all three viruses). The multiplex RT-PCR, with high sensitivity and specificity, provides a new and alternative tool for the detection of PEDV, TGEV and GAR.
文摘This review presents a reflection-type holographic memory using three-dimensional(3D)speckle-shift multiplexing. First, the schematic of the proposed memory system was described. Then,experimental demonstrations of multiplexing in plane and along the depth direction were presented. The estimated storage capacity of single layer recording was introduced and the maximum storage capacity was discussed. To increase the storage capacity, the multi-layered recording was described. In the multilayered recording, the storage capacity can be increased by appropriate arrangement of holograms in each layer.
文摘Streptococcus suis is an important zoonotic agent causing severe diseases in both pigs and humans.To date,33 serotypes of S.suis have been identified based on antigenic differences in the capsular polysaccharide.The capsular polysac-charide synthesis(cps)locus encodes proteins/enzymes that are responsible for capsular production and structure variation,which is the basis of S.suis serotyping.The cps clusters of 33 reference serotypes were published.Based on serotype-specific gene wzy in the cps gene cluster,we developed multiplex PCR(mPCR)assays for the known 33 serotypes.Seven new cps loci types were discovered,an mPCR assay for identifying them was developed in this study.The detection limit of the mPCR assay for the genomic DNA of representative strains of each new cps loci was 100 pg.Moreover,this mPCR strategy was applied in 91 untypable S.suis strains,and it turned out that 71 of them belong to these new cps types.The multiplex PCR assays deve-loped in this study provide a rapid and specific method for identification of 7 new cps types of S.suis.
文摘目的肿瘤微环境为肿瘤的发生、发展和转移提供了必要的养分和条件,而三级淋巴结构(tertiary lymphoid structures,TLS)对塑造肿瘤微环境的免疫状态非常重要。本研究为更好地实现TLS组织原位检测和分析,探讨了基于酪胺信号放大系统(tyramide signal amplification,TSA)的多重荧光染色技术在连续切片中实现TLS中多个细胞亚群的检测方案。方法选取北京大学第三医院病理科存档的6例结肠癌组织制备连续的石蜡切片。利用TSA多重荧光染色技术进行两个组合的细胞亚型标志物染色,通过图像配准及定量分析实现TLS亚群的共检测。结果两个组合检测到的荧光信号清晰无串扰,配准后可观察TLS中11个细胞亚群的分布和状态。不同样本间TLS的细胞亚群构成呈现异质性。结论该方法可同时检测TLS中的多种细胞亚群,对评估TLS状态指导临床治疗提供技术支撑。
