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    Biological fluid assay methods
    4.
    发明授权
    Biological fluid assay methods 有权
    生物流体测定方法

    公开(公告)号：US06329162B1

    公开(公告)日：2001-12-11

    申请号：US09550723

    申请日：2000-04-17

    申请人： Qinghong Han ,  Li Tang ,  Mingxu Xu ,  Yuying Tan ,  Shigeo Yagi

    发明人： Qinghong Han ,  Li Tang ,  Mingxu Xu ,  Yuying Tan ,  Shigeo Yagi

    IPC分类号： C12Q148

    CPC分类号： C12Q1/48 ,  G01N2333/91011

    摘要： A method to assess the level of folate in a biological sample comprises: providing said sample with glycine N-methyltransferase (GMT) and with an excess of S-adenosyl methionine (SAM) and of glycine; providing a control which contains no folate with said GMT and excess SAM and glycine in comparable amounts to those provided to the sample; and comparing the concentration of at least one product formed in the sample with the concentrations of said product formed in the control, whereby the difference in levels of said product in the sample as compared to the control is directly proportional to the level of folate in the sample.

    摘要翻译： 评估生物样品中叶酸水平的方法包括：向所述样品提供甘氨酸N-甲基转移酶（GMT）和过量的S-腺苷甲硫氨酸（SAM）和甘氨酸;提供不含所述叶酸的叶酸的对照 GMT和过量的SAM和甘氨酸与提供给样品的量相当; 并将样品中形成的至少一种产品的浓度与对照中形成的所述产物的浓度进行比较，由此与对照相比，样品中所述产物的水平差异与样品中叶酸水平成正比 。

    High specificity homocysteine assays for biological samples
    5.
    发明授权
    High specificity homocysteine assays for biological samples 有权
    生物样品的高特异性同型半胱氨酸测定

    公开(公告)号：US6066467A

    公开(公告)日：2000-05-23

    申请号：US340991

    申请日：1999-06-28

    申请人： Mingxu Xu ,  Yuying Tan ,  Qinghong Han ,  Li Tang

    发明人： Mingxu Xu ,  Yuying Tan ,  Qinghong Han ,  Li Tang

    IPC分类号： C12Q1/527 ,  G01N33/68 ,  C12Q1/37 ,  C12Q1/00

    CPC分类号： C12Q1/527 ,  G01N33/6815 ,  Y10S435/975

    摘要： Novel enzymatic methods to determine the concentration of homocysteine in biological fluids are described. In a typical embodiment of the invention, the biological fluid sample is from a patient, and the methods of the invention are useful to assess risk for cardiovascular disease. The novel methods of the invention involve use of particular homocysteinase enzymes that permit the determination of homocysteine concentrations in biological samples without interference from the concentrations of cysteine and/or of methionine that are routinely present in such samples. There is also provided a diagnostic kit for use in determining the amount of homocysteine in a biological sample comprising (a) a homocysteinase having the aforementioned characteristics, and (b) at least one reagent capable of being used to determine the amount of product formed in the homocysteinase reaction. In a further aspect, the homocysteinase is provided as a chimeric molecule that comprises amino acid subsequences derived from, or patterned on, more than one homocysteinase, and which is typically produced from a chimeric polynucleotide that encodes therefor. Additional enhancements in homocysteine assay methodology include use of the enzyme .gamma.-glutamylcysteine synthetase to further limit any interference from cysteine present in the biological samples.

    摘要翻译： 描述了确定生物液体中同型半胱氨酸浓度的新型酶法。 在本发明的典型实施方案中，生物流体样品来自患者，本发明的方法可用于评估心血管疾病的风险。 本发明的新方法涉及使用特异性同型半胱氨酸酶，其允许在生物样品中测定同型半胱氨酸浓度，而不受这些样品中常规存在的半胱氨酸和/或甲硫氨酸的浓度的干扰。 还提供了用于确定生物样品中同型半胱氨酸量的诊断试剂盒，其包含（a）具有上述特征的同型半胱氨酸酶，和（b）至少一种能够用于测定形成的产物量的试剂 同型半胱氨酸酶反应。 在另一方面，提供同型半胱氨酸酶作为嵌合分子，其包含衍生自或多于一种同型半胱氨酸酶的图案化的氨基酸亚序列，并且其通常由其编码的嵌合多核苷酸产生。 同型半胱氨酸测定方法的其它改进包括使用γ-谷氨酰半胱氨酸合成酶来进一步限制存在于生物样品中的半胱氨酸的任何干扰。

    Homogeneous enzymatic assay for vitamin B6 and improvements in H2S detection
    8.
    发明授权
    Homogeneous enzymatic assay for vitamin B6 and improvements in H2S detection 有权
    维生素B6的均匀酶分析和H2S检测的改进

    公开(公告)号：US06426194B1

    公开(公告)日：2002-07-30

    申请号：US09495889

    申请日：2000-02-01

    申请人： Mingxu Xu ,  Qinghong Han ,  Yuying Tan

    发明人： Mingxu Xu ,  Qinghong Han ,  Yuying Tan

    IPC分类号： C12Q137

    CPC分类号： C12Q1/527 ,  Y10S435/968 ,  Y10S435/975

    摘要： Enzymatic methods to determine the concentration of pyridoxal 5′-phosphate (PLP) in biological fluids are described. The methods of the invention are useful to assess risk for cardiovascular disease. The assay can be a homogeneous assay using the ability of PLP to function as a co-enzyme for homocysteinase and related enzymes and measuring the products of the reaction preferably spectrophotometrically. The invention also includes improvements in sensitivity of assays for measuring hydrogen sulfide production by measuring fluorescence as opposed to absorbance of the oxidized product of H2S with N,N-dialkyl p-phenylene diamine.

    摘要翻译： 描述了确定生物液体中吡哆醛5'-磷酸（PLP）浓度的酶法。 本发明的方法可用于评估心血管疾病的风险。 该测定可以是使用PLP用作同型半胱氨酸酶和相关酶的辅酶的能力的均一测定法，并且优选地通过分光光度法测量反应产物。 本发明还包括通过测量荧光来测量硫化氢产生的测定的灵敏度的改进，而不是H2S与N，N-二烷基对苯二胺的氧化产物的吸光度。