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
Red-green-blue(RGB)beam combiners are widely used in scenarios such as augmented reality/virtual reality(AR/VR),laser projection,biochemical detection,and other fields.Optical waveguide combiners have attracted extens...Red-green-blue(RGB)beam combiners are widely used in scenarios such as augmented reality/virtual reality(AR/VR),laser projection,biochemical detection,and other fields.Optical waveguide combiners have attracted extensive attention due to their advantages of small size,high multiplexing efficiency,convenient mass production,and low cost.An RGB beam combiner based on directional couplers is designed,with a core-cladding relative refractive index difference of 0.75%.The RGB beam combiner is optimized from the perspective of parameter optimization.Using the beam propagation method(BPM),the relationship between the performance of the RGB beam combiner and individual parameters is studied,achieving preliminary optimization of the device’s performance.The key parameters of the RGB beam combiner are optimized using the entropy weight-technique for order preference by similarity to an ideal solution TOPSIS method,establishing the optimal parameter scheme and further improving the device’s performance indicators.The results show that after optimization,the multiplexing efficiencies for red,green,and blue lights,as well as the average multiplexing efficiency,reached 99.17%,99.76%,96.63%and 98.52%,respectively.The size of the RGB beam combiner is 4.768 mm×0.062 mm.展开更多
为建立可同时检测甘薯褪绿矮化病毒(SPCSV)、甘薯G病毒(SPVG)和甘薯羽状斑驳病毒(SPFMV)多重RT-PCR检测方法,本文根据SPCSV热激蛋白基因(hsp70)及SPVG、SPFMV外壳蛋白基因(CP)基因核苷酸序列的保守区域设计特异性引物,通过引...为建立可同时检测甘薯褪绿矮化病毒(SPCSV)、甘薯G病毒(SPVG)和甘薯羽状斑驳病毒(SPFMV)多重RT-PCR检测方法,本文根据SPCSV热激蛋白基因(hsp70)及SPVG、SPFMV外壳蛋白基因(CP)基因核苷酸序列的保守区域设计特异性引物,通过引物筛选,优化多重RT-PCR反应条件,建立了能同时检测SPCSV、SPVG和SPFMV 3种病毒的多重RT-PCR检测方法。该体系能有效扩增出大小为304、433、601 bp 3个特异性片段。测序结果表明3种病毒与参考序列的一致性达94%-99%。应用建立的多重RT-PCR检测方法可稳定、准确、灵敏地同时检测单一或复合侵染3种甘薯病毒,为甘薯脱毒和病毒病诊断奠定基础。展开更多
【目的】建立鸡传染性支气管炎病毒(IBV)快速、准确的检测方法。【方法】根据GenBank中已经发表的793/B型和多株IBV的N基因和S1基因,对比分析后以N基因的保守序列设计合成了1对引物,跨度为582bp;以793/B的S1基因设计合成了1对特异型引物...【目的】建立鸡传染性支气管炎病毒(IBV)快速、准确的检测方法。【方法】根据GenBank中已经发表的793/B型和多株IBV的N基因和S1基因,对比分析后以N基因的保守序列设计合成了1对引物,跨度为582bp;以793/B的S1基因设计合成了1对特异型引物,跨度为891 bp;建立了能检测出IBV并能够同时鉴定出793/B血清型的实验室一次性PCR诊断方法。【结果】多重RT-PCR检测表明,793/B血清型可出现582和891 bp 2条核酸片段,而其他血清型只出现582 bp 1条核酸片段,新城疫病毒、传染性喉气管炎、鹅副粘病毒则无任何条带出现。【结论】所建立的TR-PCR方法具有一定的特异性,可用于IBV的快速检测和793/B血清型的临床诊断。展开更多
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Project(52175445)supported by the National Natural Science Foundation of ChinaProject(2022JJ30743)supported by the Natural Science Foundation of Hunan Province,China+1 种基金Project(2023GK2024)supported by the Key Research and Development Program of Hunan Province,ChinaProject(2023ZZTS0391)supported by the Fundamental Research Funds for the Central Universities of China。
文摘Red-green-blue(RGB)beam combiners are widely used in scenarios such as augmented reality/virtual reality(AR/VR),laser projection,biochemical detection,and other fields.Optical waveguide combiners have attracted extensive attention due to their advantages of small size,high multiplexing efficiency,convenient mass production,and low cost.An RGB beam combiner based on directional couplers is designed,with a core-cladding relative refractive index difference of 0.75%.The RGB beam combiner is optimized from the perspective of parameter optimization.Using the beam propagation method(BPM),the relationship between the performance of the RGB beam combiner and individual parameters is studied,achieving preliminary optimization of the device’s performance.The key parameters of the RGB beam combiner are optimized using the entropy weight-technique for order preference by similarity to an ideal solution TOPSIS method,establishing the optimal parameter scheme and further improving the device’s performance indicators.The results show that after optimization,the multiplexing efficiencies for red,green,and blue lights,as well as the average multiplexing efficiency,reached 99.17%,99.76%,96.63%and 98.52%,respectively.The size of the RGB beam combiner is 4.768 mm×0.062 mm.
文摘为建立可同时检测甘薯褪绿矮化病毒(SPCSV)、甘薯G病毒(SPVG)和甘薯羽状斑驳病毒(SPFMV)多重RT-PCR检测方法,本文根据SPCSV热激蛋白基因(hsp70)及SPVG、SPFMV外壳蛋白基因(CP)基因核苷酸序列的保守区域设计特异性引物,通过引物筛选,优化多重RT-PCR反应条件,建立了能同时检测SPCSV、SPVG和SPFMV 3种病毒的多重RT-PCR检测方法。该体系能有效扩增出大小为304、433、601 bp 3个特异性片段。测序结果表明3种病毒与参考序列的一致性达94%-99%。应用建立的多重RT-PCR检测方法可稳定、准确、灵敏地同时检测单一或复合侵染3种甘薯病毒,为甘薯脱毒和病毒病诊断奠定基础。
文摘【目的】建立鸡传染性支气管炎病毒(IBV)快速、准确的检测方法。【方法】根据GenBank中已经发表的793/B型和多株IBV的N基因和S1基因,对比分析后以N基因的保守序列设计合成了1对引物,跨度为582bp;以793/B的S1基因设计合成了1对特异型引物,跨度为891 bp;建立了能检测出IBV并能够同时鉴定出793/B血清型的实验室一次性PCR诊断方法。【结果】多重RT-PCR检测表明,793/B血清型可出现582和891 bp 2条核酸片段,而其他血清型只出现582 bp 1条核酸片段,新城疫病毒、传染性喉气管炎、鹅副粘病毒则无任何条带出现。【结论】所建立的TR-PCR方法具有一定的特异性,可用于IBV的快速检测和793/B血清型的临床诊断。