Laboratory of Seminology-Sperm Bank "Loredana Gandini", Department of Experimental Medicine, "Sapienza" University of Rome, Viale del Policlinico 155, 00161, Rome, Italy.
Laboratory of Virology, Department of Molecular Medicine, "Sapienza" University of Rome, Rome, Italy.
J Endocrinol Invest. 2021 Dec;44(12):2675-2684. doi: 10.1007/s40618-021-01580-x. Epub 2021 Apr 30.
Due to relevant repercussions on reproductive medicine, we aimed to evaluate feasibility of RT-PCR as a detection method of SARS-CoV-2 RNA in seminal fluid.
A qualitative determination of the RT-PCR assays in semen was performed through different approaches: (1) efficiency of RNA extraction from sperm and seminal plasma was determined using PRM1 and PRM2 mRNA and a heterologous system as control; (2) samples obtained by diluting viral preparation from a SARS-CoV-2 panel (virus cultured in Vero E6 cell lines) were tested; (3) viral presence in different fractions of seminal fluid (whole sample, seminal plasma and post-centrifugation pellet) was evaluated. Semen samples from mild and recovered COVID-19 subjects were collected by patients referring to the Infectious Disease Department of the Policlinico Umberto I Hospital - "Sapienza" University of Rome. Control subjects were recruited at the Laboratory of Seminology-Sperm Bank "Loredana Gandini'' of the same hospital.
The control panel using viral preparations diluted in saline and seminal fluid showed the capability to detect viral RNA presence with C values depending on the initial viral concentration. All tested semen samples were negative for SARS-CoV-2, regardless of the nasopharyngeal swab result or seminal fluid fraction.
These preliminary data show that RT-PCR for SARS-CoV-2 RNA testing appears to be a feasible method for the molecular diagnosis of SARS-CoV-2 in seminal fluid, supported by results of the control panel. The ability to detect SARS-CoV-2 in semen is extremely important for reproductive medicine, especially in assisted reproductive technology and sperm cryopreservation.
由于对生殖医学的相关影响，我们旨在评估 RT-PCR 作为检测精液中 SARS-CoV-2 RNA 的方法的可行性。
通过不同方法对精液中的 RT-PCR 检测进行定性测定：（1）使用 PRM1 和 PRM2 mRNA 以及异源系统作为对照，确定从精子和精浆中提取 RNA 的效率；（2）测试用 SARS-CoV-2 试剂盒（在 Vero E6 细胞系中培养的病毒）稀释的病毒制备物获得的样本；（3）评估精液中不同部分（全样本、精浆和离心后沉淀）的病毒存在情况。从轻度 COVID-19 患者和已康复的 COVID-19 患者收集精液样本，由患者向罗马“萨皮恩扎”大学乌姆伯托一世医院传染病科就诊收集。对照受试者由同一医院的精子银行“Loredana Gandini”的精液学实验室招募。
使用在生理盐水和精浆中稀释的病毒制剂的对照面板显示出能够检测病毒 RNA 存在的能力，C 值取决于初始病毒浓度。所有测试的精液样本均为 SARS-CoV-2 阴性，无论鼻咽拭子结果如何或精浆部分如何。
这些初步数据表明，用于 SARS-CoV-2 RNA 检测的 RT-PCR 似乎是一种可行的方法，可支持对照面板的结果，用于对 SARS-CoV-2 进行分子诊断。在精液中检测到 SARS-CoV-2 的能力对生殖医学非常重要，特别是在辅助生殖技术和精子冷冻保存中。
