Detection of HIV and HCV RNA in semen from Brazilian coinfected men using multiplex PCR before and after semen washing

Abstract

SUMMARYIntroduction. Prolonged survival of patients under HAART has resulted in new demands for assisted reproductive technologies. HIV serodiscordant couples wish to make use of assisted reproduction techniques in order to avoid viral transmission to the partner or to the newborn. It is therefore essential to test the effectiveness of techniques aimed at reducing HIV and HCV loads in infected semen using molecular biology tests.Methods. After seminal analysis, semen samples from 20 coinfected patients were submitted to cell fractioning and isolation of motile spermatozoa by density gradient centrifugation and swim-up. HIV and HCV RNA detection tests were performed with RNA obtained from sperm, seminal plasma and total semen.Results. In pre-washing semen, HIV RNA was detected in 100% of total semen samples, whereas HCV RNA was concomitantly amplified in only one specimen. Neither HIV nor HCV were detected either in the swim-up or in the post-washing semen fractions.Conclusions. Reduction of HIV and/or HCV shedding in semen by density gradient centrifugation followed by swim-up is an efficient method. These findings lead us to believe that, although semen is rarely found to contain HCV, semen processing is highly beneficial for HIV/HCV coinfected individuals.

Association of sperm apoptosis and DNA ploidy with sperm chromatin quality in human spermatozoa

“…[2] These differences may be more relevant in sperm prepared from infertile men. [3] Both TB (sperm staining blue) and %DFI may be associated with late apoptotic sperm. [4] Sperm prepared by density gradient may still have some degree of sperm DNA fragmentation.…”

“…Density-gradient centrifugation is commonly used for sperm preparation, which leads to an increased conception rate (1). This technique separates superior motile spermatozoa with normal morphology from the total sperm population, leaving behind immature, morphologically abnormal, and senescent spermatozoa with damaged DNA (2)(3)(4)(5)(6).…”

Semen processing by density gradient centrifugation is useful in selecting sperm with higher double-strand DNA integrity

“…Density gradient centrifugation is commonly used for sperm preparation that leads to an increased conception rate (Allamaneni et al , 2005). This technique separates superior motile spermatozoa with normal morphology from the total sperm population, leaving behind immature and morphologically abnormal spermatozoa (Mortimer, 1999; Henkel & Schill, 2003; Mousset‐Simeon et al , 2004; Canto et al , 2006; Morrell, 2006; Marchesi & Feng, 2007). However, there is still controversy as to the effects of this sperm separation technique on sperm cell DNA integrity.…”

Uso da técnica de swim-up para a remoção do Vírus da Artrite Encefalite Caprina do sêmen de reprodutores infectados

“…Além disso, a associação de pelo menos duas técnicas de processamento seminal e o uso de terapia antirretroviral potente apresentam excelentes resultados na eliminação das fontes ativas de transmissão do vírus e obtenção de gametas seguros (Queiroz et al, 2008). Essas informações estão de acordo com Canto et al (2006) A diminuição observada para esses parâmetros, provavelmente, estaria relacionada com o procedimento de centrifugação a que as amostras foram submetidas. Além disso, segundo Correa et al, (1997), o estresse relacionado ao tempo necessário para recuperação dos espermatozoides também pode contribuir para a elevada taxa de mortalidade e perda de capacitação dos mesmos.…”