Male fertility disorders: RhyMA Test IVFAZF a multifactorial genetic analysis.

Infertility is a major health problem affecting 10-15% of couples seeking to have children and a male factor can be identifi ed in about half of these cases. A signifi cant proportion of infertile males are affected either by oligozoospermia (reduced sperm production) or azoospermia (lack of any sperm in the ejaculate) (1).

The alteration of spermatogenesis can be the consequence of many causes but however 50% of infertile cases are unknown. Recently the growth of assisted reproduction techniques contributed to such research and the study of Y chromosome microdeletions into AZF regions shows their connection to non obstructive azoospermia (1,2); men with deletions in AZFa and AZFb regions, show severe defects in spermatogenesis, whereas deletions of AZFc region can be compatible with residual spermatogenesis (3).

The screening of AZF microdeletions results important to prevent their vertical transmission to sons in particular after the development of new reproduction techniques, that overcome spermatogenic defects and the biological selection (2).

A multiparametric analysis of the most important polymorphisms is the EUROCLONE concept for diagnosis of infertility due to genetic disorders.

1. Foresta C., Moro E., Ferlin A. “Y Chromosome Microdeletions and Alterations of Spermatogenesis” Endocrine Reviews 22(2):226-239.2. Dohle GR Halley DJJ Van Hemel JO van den Ouwel AMW Pieters MHEC Weber RFA Govaerts LCP 2002 “Genetic risk factors in infertile men with severe oligozoospermia and azoospermia.” Human Reproduction 17 13-16.3. Sertic J, Cvitkovic P, Myers A, Saiki R.K., Stavljenic Rukavina A “Genetic Markers of Male Infertility: Y Chromosome Microdeletions and Cystic Fibrosis Transmembrane Conductance Gene Mutations”. CMJ 42(4):416-420,2001.

PR 636 flyer rhyma test IVF AZF.1 1 19-09-2007 18:00:51

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Male fertility disorders: RhyMA Test IVFAZF a multifactorial genetic analysis

RhyMA Test – IVFAZF (COD. EES003020)**CE-IVD marked

Intended Use:

RhyMA Test–IVFAZF is a multiparametric detection of of the Y Chromosome microdeletions associated to male infertility, that are located into regions AZFa, AZFb, AZFc. The test has been developed in agreement with guidelines of European Molecular Genetics Quality Network (EMQN) 2001.

The kit detects the microdeletions of the Sequence Tagged Sites (STSs) into the AZF regions, see below, and the presence of SRY and ZFY genes:

Assay Principle

RhyMA Test is based on the Reverse Hybridization technology and needs four steps:• DNA extraction (not included in the kit)• Multiplex DNA amplifi cation with specifi c biotinylated primers (included in the kit)• Hybridization of amplifi ed and biotinylated DNA fragments with strips (included in the kit)• Developing reaction with biotin-streptavidine system (included in the kit).

Interpretation of results

For interpretation of results compare the strip of your sample with the DECODER Table (Fig. 1) alligning the red marked line. Results are in Table 1.

Table 1.

Genotype Identifi cation

Wild Type (Wild Type (for one or more AZF regions)

Presence of both the two lines of the linked STS

Microdeleted (fWild Type (for one or more AZF regions)

Absence of both the two lines of the linked STS

AZFa region

AZFb region

AZFc region

SRY gene

ZFY gene

sY84 sY127 sY254

sY86 sY134 sY255

Fig.1

PR 636 flyer rhyma test IVF AZF.2 2 19-09-2007 18:00:58