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    • 2. 发明申请
    • DETECTION OF BIOAGENTS USING A SHEAR HORIZONTAL SURFACE ACOUSTIC WAVE BIOSENSOR
    • 使用剪切水平表面声波生物传感器检测生物剂
    • WO2009005542A3
    • 2009-04-16
    • PCT/US2008001721
    • 2008-02-08
    • STC UNMSANDIA CORPLARSON RICHARD SHJELLE BRIANHALL PAM RBROWN DAVID CBISOFFI MARCOBROZIK SUSAN MBRANCH DARREN WEDWARDS THAYNE LWHEELER DAVID
    • LARSON RICHARD SHJELLE BRIANHALL PAM RBROWN DAVID CBISOFFI MARCOBROZIK SUSAN MBRANCH DARREN WEDWARDS THAYNE LWHEELER DAVID
    • C12Q1/34C12M3/00
    • G01N33/54373C12Q1/6825G01N29/032G01N33/553G01N2291/0422
    • Viruses and other bioagents are of high medical and biodefense concern and their detection at concentrations well below the threshold necessary to cause health hazards continues to be a challenge with respect to sensitivity, specificity, and selectivity. Ideally, assays for accurate and real time detection of viral agents and other bioagents would not necessitate any pre-processing of the analyte, which would make them applicable for example to bodily fluids (blood, sputum) and man-made as well as naturally occurring bodies of water (pools, rivers). We describe herein a robust biosensor that combines the sensitivity of surface acoustic waves (SAW) generated at a frequency of 325 MHz with the specificity provided by antibodies and other ligands for the detection of viral agents. In preferred embodiments, a lithium tantalate based SAW transducer with silicon dioxide waveguide sensor platform featuring three test and one reference delay lines was used to adsorb antibodies directed against Coxsackie virus B4 or the negative-stranded category A bioagent Sin Nombre virus (SNV), a member of the genus Hantavirus, family Bunyaviridae, negative-stranded RNA viruses. Rapid detection (within seconds) of increasing concentrations of viral particles was linear over a range of order of magnitude for both viruses, although the sensor was approximately 50x104 -fold more sensitive for the detection of SNV. For both pathogens, the sensor's selectivity for its target was not compromised by the presence of confounding Herpes Simplex virus type 1. The biosensor was able to detect SNV at doses lower than the load of virus typically found in a human patient suffering from hantavirus cardiopulmonary syndrome (HCPS). Further, in a proof-of-principle real world application, the SAW biosensor was capable of selectively detecting SNV agents in complex solutions, such as naturally occurring bodies of water (river, sewage effluent) without analyte pre-processing.
    • 病毒和其他生物制剂对医学和生物防御有很高的关注度,并且它们的检测浓度远低于引起健康危害所必需的阈值,这仍然是灵敏度,特异性和选择性方面的挑战。 理想情况下,准确和实时检测病毒剂和其他生物剂的分析不需要对分析物进行任何预处理,这将使其适用于例如体液(血液,痰液)和人造以及自然发生的 水体(水池,河流)。 我们在此描述了一种鲁棒的生物传感器,其将频率为325MHz的表面声波(SAW)的灵敏度与由抗体和其他配体提供的用于检测病毒剂的特异性相结合。 在优选的实施方案中,具有三个测试和一个参考延迟线的具有二氧化硅波导传感器平台的基于钽酸锂的SAW换能器被用于吸附针对柯萨奇病毒B4或负链A类生物因子Sin Nombre病毒(SNV)的抗体,a 汉坦病毒属,布尼亚病毒科,负链RNA病毒成员。 对于两种病毒,增加浓度的病毒颗粒的快速检测(数秒内)在一定数量级范围内是线性的,尽管对于检测SNV而言传感器的灵敏度约为50×10 4倍。 对于这两种病原体而言,传感器对其靶标的选择性不会由于混杂的1型单纯性疱疹病毒的存在而受到损害。生物传感器能够以比通常在患有汉坦病毒心肺综合征的人类患者中发现的病毒载量低的剂量检测SNV (HCPS)。 此外,在原理验证的实际应用中,SAW生物传感器能够选择性检测复杂溶液中的SNV试剂,例如没有分析物预处理的天然存在的水体(河流,污水流出物)。
    • 3. 发明申请
    • DETECTION OF BIOAGENTS USING A SHEAR HORIZONTAL SURFACE ACOUSTIC WAVE BIOSENSOR
    • 使用剪切水平表面声波生物传感器检测生物
    • WO2009005542A2
    • 2009-01-08
    • PCT/US2008/001721
    • 2008-02-08
    • STC.UNMSANDIA CORPORATIONLARSON, Richard, S.HJELLE, BrianHALL, Pam, R.BROWN, David, C.BISOFFI, MarcoBROZIK, Susan, M.BRANCH, Darren, W.EDWARDS, Thayne, L.WHEELER, David
    • LARSON, Richard, S.HJELLE, BrianHALL, Pam, R.BROWN, David, C.BISOFFI, MarcoBROZIK, Susan, M.BRANCH, Darren, W.EDWARDS, Thayne, L.WHEELER, David
    • C12Q1/70
    • G01N33/54373C12Q1/6825G01N29/032G01N33/553G01N2291/0422
    • Viruses and other bioagents are of high medical and biodefense concern and their detection at concentrations well below the threshold necessary to cause health hazards continues to be a challenge with respect to sensitivity, specificity, and selectivity. Ideally, assays for accurate and real time detection of viral agents and other bioagents would not necessitate any pre-processing of the analyte, which would make them applicable for example to bodily fluids (blood, sputum) and man-made as well as naturally occurring bodies of water (pools, rivers). We describe herein a robust biosensor that combines the sensitivity of surface acoustic waves (SAW) generated at a frequency of 325 MHz with the specificity provided by antibodies and other ligands for the detection of viral agents. In preferred embodiments, a lithium tantalate based SAW transducer with silicon dioxide waveguide sensor platform featuring three test and one reference delay lines was used to adsorb antibodies directed against Coxsackie virus B4 or the negative-stranded category A bioagent Sin Nombre virus (SNV), a member of the genus Hantavirus, family Bunyaviridae , negative-stranded RNA viruses. Rapid detection (within seconds) of increasing concentrations of viral particles was linear over a range of order of magnitude for both viruses, although the sensor was approximately 50x10 4 -fold more sensitive for the detection of SNV. For both pathogens, the sensor's selectivity for its target was not compromised by the presence of confounding Herpes Simplex virus type 1. The biosensor was able to detect SNV at doses lower than the load of virus typically found in a human patient suffering from hantavirus cardiopulmonary syndrome (HCPS). Further, in a proof-of-principle real world application, the SAW biosensor was capable of selectively detecting SNV agents in complex solutions, such as naturally occurring bodies of water (river, sewage effluent) without analyte pre-processing.
    • 病毒和其他生物制剂具有高度的医学和生物防治问题,其浓度远远低于引起健康危害的阈值的检测在敏感性,特异性和选择性方面仍然是一个挑战。 理想情况下,用于准确和实时检测病毒试剂和其他生物标签的测定不需要对分析物进行任何预处理,这将使它们适用于例如体液(血液,痰)和人造的以及天然存在的 水体(游泳池,河流)。 我们在这里描述了一种强大的生物传感器,其结合了在325MHz频率下产生的表面声波(SAW)的灵敏度与由抗体和其他配体提供的用于检测病毒剂的特异性。 在优选实施例中,使用具有三个测试和一个参考延迟线的具有二氧化硅波导传感器平台的基于钽酸锂的SAW换能器来吸附针对柯萨奇病毒B4或负链A类生物剂Sin Nombre病毒(SNV)的抗体, 汉坦病毒属,布尼亚病毒科,负链RNA病毒的成员。 尽管传感器对于SNV检测的敏感度大约为50x104倍,但对于两种病毒而言,增加浓度的病毒颗粒的快速检测(在几秒钟内)是线性范围的数量级。 对于两种病原体,传感器对其目标的选择性不受1型混合型单纯疱疹病毒的存在的影响。生物传感器能够以低于患有汉坦病毒心肺综合征的人类患者的病毒载量的剂量来检测SNV (HCPS)。 此外,在原理现实应用中,SAW生物传感器能够选择性地检测复杂解决方案中的SNV试剂,例如天然存在的水体(河流,污水流出物),无需分析物预处理。