Molecular diagnosis of SARS-CoV-2 in seminal fluid

Abstract

Purpose 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. Methods 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. Results The control panel using viral preparations diluted in saline and seminal fluid showed the capability to detect viral RNA presence with Ct 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. Conclusion 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.

Andrological effects of SARS-Cov-2 infection: a systematic review and meta-analysis

“…Paoli et al [ 43 ] did not find SARS-CoV-2 mRNA in seminal fluid and urine in 31-year-old men in the recovered phase of the infection. The same authors confirmed this result in two mild COVID-19 patients in the acute phase with a positive nasopharyngeal swab and in recovered patients with a negative swab [ 44 ]. Similar results were reported by Pan et al [ 14 ].…”

Could SARS‐CoV‐2 infection affect male fertility and sexuality?

“…ACE-2 was expressed in germ cells, Sertoli cells and Leydig cells. ACE-2 expression was higher in patients aged 20-30 while was very low in patients aged 60 [13] Semen of patients recovering from COVID-19 SARS-CoV-2 was not detected in semen after a median of 31 days from COVID-19 diagnosis [25] Semen of patients recovering from COVID-19 SARS-CoV-2 was not detected in semen [26] Semen of COVID-19 patients SARS-CoV-2 was not detected in semen or urine [27][28][29][30][31] Urine, expressed prostatic secretions (EPSs) and semen samples COVID-19 patients…”

COVID-19 and Male Reproduction: A Thorny Problem

“…Whether SARS-CoV-2 is present in the testis and semen samples of COVID-19 patients remains debatable ( Holtmann et al, 2020 ; Ma et al, 2021 ). A previous study reported that reverse transcription polymerase chain reaction (RT-PCR) is a feasible molecular diagnostic method for detecting SARS-CoV-2 RNA in semen samples ( Paoli et al, 2021 ), and SARS-CoV-2 has been actually detected in the semen samples of patients infected with SARS-CoV-2. However, most reports in the current literature have indicated that SARS-CoV-2 is undetectable in the semen samples of male COVID-19 patients.…”