Development and evaluation of a qualitative reverse-transcriptase nested polymerase chain reaction protocol for same-day viral validation of human immunodeficiency virus type 1 ribonucleic acid in processed semen

Lesage, Benoît; Vannin, Anne-Sophie; Emiliani, Serena; Debaisieux, Laurent; Englert, Yvon; Liesnard, Corinne

doi:10.1016/j.fertnstert.2005.12.021

Cited by 11 publications

(9 citation statements)

References 19 publications

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“…This method allows detection of low viral load and controls for possible PCR inhibitors. By this approach two motile spermatozoa preparations, obtained after Percoll gradient centrifugation followed by swim-up, were found to be HIV-1 RNA positive (Lesage et al, 2006). We have developed experimental strategies to remove 'contaminating' NSC from the semen, monitor any residual presence in the purified sperm fractions and verify the presence of HIV-1 proviral DNA by nested PCR, with a detection limit of 4 HIV-1 DNA copies/10 6 spermatozoa.…”

Section: Introductionmentioning

confidence: 99%

HIV-1 viral DNA is present in ejaculated abnormal spermatozoa of seropositive subjects

et al. 2007

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Section: Introductionmentioning

confidence: 99%

HIV-1 viral DNA is present in ejaculated abnormal spermatozoa of seropositive subjects

et al. 2007

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“…The effective isolation of sperm from seminal plasma and white blood cells that serve as target cells for the virus, should then yield a population of sperm that is HIV-1 free and safe for use during ART. The promotion of semen washing procedures consequently results in more HIV-1 sero-discordant couples attending infertility units, rather than attempting to conceive by having unprotected intercourse (Lesage et al, 2006). Semen decontamination procedures employed at RBL consist of semen processing by discontinuous density gradient centrifugation in combination with the ProInsert™.…”

Section: Discussionmentioning

confidence: 99%

“…ii) The presence of seminal PCR inhibitors results in the inhibition of HIV-1 RNA amplification whereby viral validation errors are obtained (Lesage et al, 2006). Patients should be informed that in the event of PCR errors, viral validation should be repeated, whereby less sperm will be available for ART.…”

Section: Discussionmentioning

confidence: 99%

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Semen decontamination for the elimination of seminal HIV-1

Fourie

Loskutoff²,

Huyser

2015

Reproductive BioMedicine Online

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The risk of HIV transmission to the female partner, or potential offspring of an HIV-1 infected male can be reduced using semen decontamination procedures prior to assisted reproductive treatment (ART). The objective of this study was to determine the efficiency to decontaminate semen samples (N=186) from 95 HIV-1 sero-positive patients. Aliquots of neat semen were submitted for viral validation by qualitative and quantitative polymerase chain reaction. Semen samples were processed by density gradient centrifugation in combination with a ProInsert™ tube where after aliquots of the processed sperm samples were analysed for the presence of HIV-1. Fifty four percent of all tested neat semen samples tested positive for HIV-1 DNA, RNA or both (13.4%, 11.3% and 29.0%, respectively). From a total of 103 processed sperm samples that were submitted for viral validation, two samples tested positive for HIV-1 DNA and none for RNA. In conclusion, semen processing with the ProInsert™ followed by viral validation of processed sperm samples should be performed when providing assisted reproductive treatment to couples where the male partner is HIV-1 seropositive.

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“…Developments in highly sensitive polymerase chain reaction techniques have recently demonstrated that low viral loads of HIV-1 can still be present in samples processed through gradient columns where all nonmotile cells have been removed [17]. For this reason, it is important to continue developing techniques to validate the absence of HIV and other transmissible viruses in semen samples processed for IUI [18].…”

Section: Infectious Disease Processingmentioning

confidence: 99%

Sperm Preparation for Artificial Insemination

Christensen

2010

Reproductive Endocrinology and Infertility

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Intrauterine insemination (IUI) is a common treatment for couples with infertility issues. In order to operate a good IUI program, several elements are necessary. This chapter attempts to provide a comprehensive description of these elements including: aspects of patient education, communication between the laboratory and the referring physician, employee training, safety, sample identification, and documentation procedures. It also describes supplies and reagents needed for IUI, multiple techniques for the preparation of sperm, and advantages of the different preparations. A discussion of special considerations such as the use of positive samples for infectious disease, use of sex-selected sperm, and chemical motility enhancement is included. Keywords IntroductionDue to increased social acceptance and the successful range of infertility therapies now available, more people than ever are seeking treatment for infertility. A broad range of conditions, affecting both the male and female, contribute to infertility. Many circumstances, such as severe oligozoospermia or blocked fallopian tubes, are immediate indications that more aggressive therapies, such as in vitro fertilization (IVF), are needed to be pursued for a reasonable chance of success. However, the majority of patients with less severe or unexplained forms of infertility have a reasonable chance of success with less invasive intrauterine insemination (IUI). While the decision of which infertility therapy to pursue depends on a number of factors including infertile pathologies, age, and finances, for many couples IUI will be the first and only therapy needed.Several factors can contribute to the success rates of IUI, including sperm quality, insemination timing and number of inseminations, age, and the use of ovarian stimulation [1][2][3]. One of the major male factors influencing the rates is the concentration of motile sperm in the prepared sample used for insemination, with pregnancy rates falling off quickly when less than 5 million motile sperm are inseminated [4,5]. Normal sperm morphology has also been evaluated, though the correlation with pregnancy rates is less clear. On average, the pregnancy rate per IUI cycle is about 10-15% when motile sperm are available, though higher rates have been reported, especially when combined with the use of gonadotropins [6].Under normal circumstances, sperm must pass through the cervical mucus to reach the uterus, a process that limits uterine entrance to only the healthiest sperm and prevents dead sperm, debris, and seminal fluid from getting in. Due to the presence of prostaglandins and microbes in the seminal fluid, semen cannot be placed directly into the uterus without the potential of inducing painful cramping and infection. With the advent of sperm preparation techniques to remove seminal fluids, nonmotile cells, and debris, sperm can be placed directly into the uterus with minimal risk. In instances where sperm quality is less than optimal or adverse mucus or cervical conditions exist, IUI can b...

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Development and evaluation of a qualitative reverse-transcriptase nested polymerase chain reaction protocol for same-day viral validation of human immunodeficiency virus type 1 ribonucleic acid in processed semen

Cited by 11 publications

References 19 publications

HIV-1 viral DNA is present in ejaculated abnormal spermatozoa of seropositive subjects

HIV-1 viral DNA is present in ejaculated abnormal spermatozoa of seropositive subjects

Semen decontamination for the elimination of seminal HIV-1

Sperm Preparation for Artificial Insemination

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