Method for quantification of a prostate cancer biomarker in urine without sample preparation
Method for quantification of a prostate cancer biomarker in urine without sample preparation
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DOI：
10.1021/ac070895z
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发表时间：
2007-10-15
影响因子：
7.4
通讯作者：
Mutharasan, Raj
中科院分区：
文献类型：
--
作者：
Maraldo, David;Garcia, Fernando U.;Mutharasan, Raj
我们描述了一种基于大悬臂的方法，可以直接从患者尿液中检测前列腺癌生物标记物(α-甲基酰辅酶A消旋酶；AMACR)，而不需要样品制备步骤和标记试剂的使用。5例确诊前列腺癌患者于活检后3周前瞻性采集清洁尿液标本。将3mLAMACR暴露在固定了AMACR的压电式毫米波(PEMC)传感器上，以盲法检测AMACR的存在。当样品中的AMACR与传感器上的抗体结合时，PEMC的共振频率降低。5例患者的共振频率变化分别为4,1314+/非35超集(n=2)，269+/非超集17(n=2)，977+/非超集(n=3)，600+/非超集31(n=2)和801+/非超集81(n=2)赫兹。在15min内即可观察到阳性检测。对阳性、阴性和缓冲对照的反应分别是-9+/非13超集、-34+/非18超集和-6+/非18赫兹超集。低pH缓冲液释放证实了AMACR附着的阳性验证。传感器的响应与对照尿中的AMACR浓度有定量关系，并利用这种关系建立了一种原位校准方法来定量测定患者尿中的AMACR。三名患者的尿中AMACR的估计浓度分别为42、2和3fg/mLAMACR，而对照尿中证实没有AMACR(n=13)。由于组合测量的简单性。由于具有高度的敏感性和特异性，该方法可能是在护理地点环境中识别前列腺癌风险增加的男性的有用辅助工具。
We describe a macrocantilever-based method for detecting a prostate cancer biomarker (alpha-methylacyl-CoA racemase; AMACR) directly in patient urine without a sample preparation step and without the use of labeled reagents. Clean catch voided urine specimens were prospectively collected from five confirmed prostate cancer patients 3 weeks postbiopsy. The presence of AMACR was measured in a blinded manner by exposing 3 mL of urine to the antiAMACR-immobilized piezoelectric-excited millimeter-sized (PEMC) sensor. The resonance frequency of PEMC decreases as AMACR from sample binds to the antibody on the sensor. The resonance frequency changes for the five patients tested were 4,1314 +/not superset of 35 (n = 2), 269 +/not superset of 17 (n = 2), 977 +/not superset of 64 (n = 3), 600 +/not superset of 31 (n = 2), and 801 +/not superset of 81 (n = 2) Hz, respectively. Positive detection was observed within similar to 15 min. The responses to positive, negative, and buffer controls were -9 +/not superset of 13, -34 +/not superset of 18, and -6 +/not superset of 18 Hz, respectively. Positive verification of AMACR attachment was confirmed by low-pH buffer release. The sensor response was quantitatively related to AMACR concentration in control urine, and the relationship was used in developing an in situ calibration method for quantifying AMACR in patient urine. Estimated concentrations of 42, 2, and 3 fg/mL AMACR were calculated for the three patients' urine, while absence of AMACR was confirmed in control urine (n = 13). Because of simplicity of measurement combined. with high sensitivity and specificity, the method may be a useful adjunct in a point-of-care setting to identify men at increased risk for prostate cancer.