Multiple recognition assay reveals prostasomes as promising plasma biomarkers for prostate cancer
Multiple recognition assay reveals prostasomes as promising plasma biomarkers for prostate cancer
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DOI：
10.1073/pnas.1019330108
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发表时间：
2011-05-24
影响因子：
11.1
通讯作者：
Kamali-Moghaddam, Masood
中科院分区：
文献类型：
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作者：
Tavoosidana, Gholamreza;Ronquist, Gunnar;Kamali-Moghaddam, Masood
前列腺小体是由正常和恶性前列腺腺泡细胞产生和分泌的微泡（平均直径150纳米）。据推测，恶性前列腺细胞的侵袭性生长可能导致这些通常释放到精液中的微泡出现在间质间隙，从而进入外周循环。前列腺小体作为前列腺癌患者血液生物标志物的适用性通过一种扩展的近端结扎试验（PLA）进行了测试。我们开发了一种非常敏感和特异性的检测方法（4PLA），用于检测复杂的目标结构，如微泡，其中目标首先通过固定抗体捕获，随后通过使用附加DNA链的其他四种抗体检测。需要五种抗体同时结合才能产生可扩增的报告基因，从而增加了特异性和敏感性。与对照组和活检结果良性的男性相比，该检测成功地检测到根治性前列腺切除术前前列腺癌患者血液样本中前列腺体水平显著升高。在两项独立研究中，前列腺癌患者血浆中前列腺体水平的中位数比对照样本高2.5至7倍，该检测还区分了前列腺切除术Gleason评分高和中等的患者（分别为8/9和7）和低评分的患者（分别为8/9和7）。
Prostasomes are microvesicles (mean diameter, 150 nm) that are produced and secreted by normal and malignant prostate acinar cells. It has been hypothesized that invasive growth of malignant prostate cells may cause these microvesicles, normally released into seminal fluid, to appear in interstitial space and therewith into peripheral circulation. The suitability of prostasomes as blood biomarkers in patients with prostate cancer was tested by using an expanded variant of the proximity ligation assay (PLA). We developed an extremely sensitive and specific assay (4PLA) for detection of complex target structures such as microvesicles in which the target is first captured via an immobilized antibody and subsequently detected by using four other antibodies with attached DNA strands. The requirement for coincident binding by five antibodies to generate an amplifiable reporter results in both increased specificity and sensitivity. The assay successfully detected significantly elevated levels of prostasomes in blood samples from patients with prostate cancer before radical prostatectomy, compared with controls and men with benign biopsy results. The medians for prostasome levels in blood plasma of patients with prostate cancer were 2.5 to sevenfold higher compared with control samples in two independent studies, and the assay also distinguished patients with high and medium prostatectomy Gleason scores (8/9 and 7, respectively) from those with low score (