Dubois C, Figarella-Branger D, Rougon G, Rampini C
Laboratoire de Biologie Cellulaire II, Faculté de Médecine Saint-Antoine, Paris, France.
C R Seances Soc Biol Fil. 1998;192(2):289-96.
We described a double-site enzyme-linked immunosorbent assay (ELISA) to measure polysialic acid neural cell adhesion molecule (PSA-NCAM) level in CSF. Immunocapture of PSA-bearing molecules is first effected by means of a monoclonal antibody (anti-MenB), directed against sialic acid polymers and adsorbed into plastic wells. Linked PSA-NCAM is then revealed by means of a second antibody, directed against an aminoacid sequence of NCAM and labelled with peroxydase. The lowest amount of PSA-NCAM detectable was estimated to be 0.11 microgram/l. This value was considered as the threshold for positivity. PSA-NCAM level was measured using this method in CSF from 29 patients with medulloblastoma. CSF had been collected at different times following tumor excision and stored at--80 degrees C. At the same times, cytological examination in CSF (medulloblastoma metastatic cells) and craniospinal imaging (tomographic scan or MRI) had been performed. PSA-NCAM was never detected in control CSF. For patients in remission, beyond the post-operative period of 1 or 2 months, 18 on 21 exhibited a PSA-NCAM level below the threshold value. For refractory patients, so classified according to the positivity of cytology and/or imaging, whatever the time after the tumor excision, PSA-NCAM was always positive (23/23), while either cytology or imaging were positive less frequently (16/23 for both). For relapses, PSA-NCAM was more frequently positive (6/7) than cytology and imaging (1/7 and 5/7, respectively). We concluded that PSA-NCAM positivity in CSF may be a reliable marker to detect the invasive or metastatic feature of medulloblastoma.
我们描述了一种双位点酶联免疫吸附测定法(ELISA),用于测量脑脊液中多唾液酸神经细胞黏附分子(PSA-NCAM)的水平。首先,通过针对唾液酸聚合物的单克隆抗体(抗-MenB)进行免疫捕获,该抗体吸附于塑料孔中,以捕获携带PSA的分子。然后,通过针对NCAM氨基酸序列的第二抗体进行检测,该抗体用过氧化物酶标记,以显示连接的PSA-NCAM。可检测到的PSA-NCAM的最低量估计为0.11微克/升。该值被视为阳性阈值。使用该方法对29例髓母细胞瘤患者的脑脊液进行了PSA-NCAM水平检测。脑脊液在肿瘤切除后的不同时间收集,并储存在-80℃。同时,对脑脊液进行了细胞学检查(髓母细胞瘤转移细胞)和颅脊髓成像(断层扫描或MRI)。在对照脑脊液中从未检测到PSA-NCAM。对于缓解期患者,在术后1或2个月之后,21例中有18例的PSA-NCAM水平低于阈值。对于难治性患者,根据细胞学和/或成像的阳性结果进行分类,无论肿瘤切除后的时间如何,PSA-NCAM始终为阳性(23/23),而细胞学或成像阳性的频率较低(两者均为总例数的16/23)。对于复发患者,PSA-NCAM阳性的频率(6/7)高于细胞学和成像(分别为1/7和5/7)。我们得出结论,脑脊液中PSA-NCAM阳性可能是检测髓母细胞瘤侵袭或转移特征的可靠标志物。