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精子的荧光原位杂交技术:斑点计数能否取代传统计数?目前还不行。

FISH on sperm: spot-counting to stop counting? Not yet.

作者信息

Molina Òscar, Sarrate Zaida, Vidal Francesca, Blanco Joan

机构信息

Unitat de Biologia Cel·lular (Facultat de Biociències), Universitat Autònoma de Barcelona, Bellaterra (Cerdanyola del Vallès), Spain.

Unitat de Biologia Cel·lular (Facultat de Biociències), Universitat Autònoma de Barcelona, Bellaterra (Cerdanyola del Vallès), Spain.

出版信息

Fertil Steril. 2009 Oct;92(4):1474-1480. doi: 10.1016/j.fertnstert.2008.07.1779. Epub 2008 Oct 17.

Abstract

OBJECTIVE

To evaluate the reliability and applicability of the spot-counting system (Cytovision Spot AX workstation) which offers an alternative to the tedious manual analysis of sperm fluorescence in situ hybridization (FISH).

DESIGN

Manual and automatic analyses were performed and compared.

SETTING

Universitat Autònoma de Barcelona.

PATIENT(S): Twenty-four men who asked for information on infertility showing different seminal parameters.

INTERVENTION(S): A semen sample for each patient was collected and prepared for FISH.

MAIN OUTCOME MEASURE(S): A dual-color FISH using specific probes for chromosomes 13 and 21 and a triple-color FISH with centromeric probes for chromosomes 18, X, and Y were used (Vysis). Standard FISH analysis was carried out. Automatic analysis was subsequently performed using a Spot AX system.

RESULT(S): Overall, we performed 120 comparisons. In 116 out of 120 (96.67%), the percentage of anomalies reported using manual counting fell within the incidence detected using the automatic system. In the remaining comparisons, statistical differences were detected (4 out of 120; 3.33%). Time consumed by the automatic analysis was always higher than the manual one, being influenced by the characteristics of the preparations.

CONCLUSION(S): The spot-counting system has potential, but before the service is ready to be offered, we still need to overcome some limitations associated with it.

摘要

目的

评估斑点计数系统(赛托维斯斑点AX工作站)的可靠性和适用性,该系统为精子荧光原位杂交(FISH)繁琐的手动分析提供了一种替代方法。

设计

进行手动和自动分析并比较。

地点

巴塞罗那自治大学。

患者

24名询问不育症信息的男性,其精液参数各不相同。

干预措施

为每位患者采集一份精液样本并准备进行FISH检测。

主要观察指标

使用针对13号和21号染色体的特异性探针进行双色FISH检测,以及使用针对18号、X和Y染色体的着丝粒探针进行三色FISH检测(Vysis公司产品)。进行标准FISH分析。随后使用斑点AX系统进行自动分析。

结果

总体而言,我们进行了120次比较。在120次比较中的116次(96.67%)中,手动计数报告的异常百分比落在自动系统检测到的发生率范围内。在其余比较中,检测到统计学差异(120次中的4次;3.33%)。自动分析所花费的时间总是高于手动分析,这受到样本制备特点的影响。

结论

斑点计数系统具有潜力,但在准备提供该服务之前,我们仍需要克服与之相关的一些局限性。

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