Monteiro Rosa Alice Casemiro, Pariz Juliana Risso, Pieri Patricia de Campos, Hallak Jorge
Androscience - Pesquisa Clínica de alta Complexidade e Laboratório de Andrologia, São Paulo, Brasil.
Seção de Andrologia - Divisão de Urologia, Faculdade de Medicina da Universidade de São Paulo, São Paulo, Brasil.
Int Braz J Urol. 2016 Jan-Feb;42(1):132-8. doi: 10.1590/S1677-5538.IBJU.2015.0090.
This study describes a new method of microcentrifugation as an improved, viable, cost-effective option to the classical Cytospin apparatus to confirm azoospermia. Azoospermic semen samples were evaluated for cryptozoospermia by a centrifugation method similar to that of World Health Organization guidelines (2010; entire specimen centrifuged at 3000g for 15 min, and aliquots of the pellet examined). Then, if no sperm were detected, the pellet from that procedure was resuspended in culture medium, centrifuged (2000g for 15 min), and the entire pellet spread on a 4 X 6mm area of a slide and stained using the Christmas tree method (Nuclear-Fast solution and picric acid). The entire stained area was examined for the presence or absence of sperm. A total of 148 azoospermic samples (after standard WHO diagnosis) were included in the study and 21 samples (14.2%) were identified as sperm-positive. In all microcentrifugation slides, intact spermatozoa could be easily visualized against a clear background, with no cellular debris. This novel microcentrifugation technique is clearly a simple and effective method, with lower cost, increasing both sensitivity and specificity in confirming the absence or presence of spermatozoa in the ejaculate. It may represent a step forward of prognostic value to be introduced by andrology laboratories in the routine evaluation of patients with azoospermia in the initial semen analysis.
本研究描述了一种新型微量离心法,作为一种经过改进的、可行的、具有成本效益的方法,可替代传统的细胞离心涂片仪来确诊无精子症。采用一种类似于世界卫生组织指南(2010年;将整个标本以3000g离心15分钟,检查沉淀物的等分试样)的离心方法,对无精子症精液样本进行隐匿精子症评估。然后,如果未检测到精子,则将该步骤得到的沉淀物重悬于培养基中,离心(2000g,15分钟),将整个沉淀物铺展在载玻片4×6mm的区域上,并用圣诞树法(核固红溶液和苦味酸)染色。检查整个染色区域有无精子。本研究共纳入148份无精子症样本(经过世界卫生组织标准诊断),其中21份样本(14.2%)被鉴定为精子阳性。在所有微量离心涂片上,完整的精子在清晰的背景下易于观察到,且无细胞碎片。这种新型微量离心技术显然是一种简单有效的方法,成本较低,在确认射精中精子的有无方面提高了敏感性和特异性。它可能代表了男科学实验室在无精子症患者初始精液分析的常规评估中引入的具有预后价值的一个进步。