INTRODUCTION: Polar body (PB) biopsy prior to oocyte insemination, with subsequent genetic analysis, is an alternative approach to Preimplantation Genetic Diagnosis (PGD) for the assessment of maternal genetic inheritance. It might be considered an ethically acceptable way to perform genetic testing for couples with moral objections to discard embryos. Additionally, in Italy, where the law imposes strict limitations, the only option is the first polar body (1stPB) testing, called Pre-Conception Genetic Diagnosis (PCGD). In PCGD, the ICSI insemination can be performed only after genetic diagnosis of oocytes. There is a narrow window of time available for PCGD in order to prevent oocyte in-vitro ageing. This objective of the current study was to evaluate laboratory and clinical outcomes of PCGD carried out in 21 cases. We performed this technique in two different ways for remove the 1stPB. In addition, different access points of injection during ICSI were adopted. MATERIALS AND METHODS: Twenty-one PCGD cycles, were performed from December 2007 to January 2009. Couples were divided in two groups. In group A (N=8), 1stPB removal was performed applying a laser microdissection at 1 o’clock respect to the PB positioned at 12 o’clock (tangential way). In group B (N=13), laser zona opening was applied in the same position of the 1stPB, placed at 3 o’clock (frontal way). In group A, unaffected oocytes were inseminated with conventional ICSI (injection at 3 o'clock respect the position of 1stPB removed). In group B, the injection pipette was introduced throw the hole previously created to remove the PB. After the biopsy procedure all 1stPBs were sent to the genetic laboratory. The analysis was performed in 4-5 hours by a PCR technique. All unaffected oocytes, were inseminated from 43 to 45 hours after HCG administration. Statistical analysis was performed using the Mann-Whitney and the Fisher’s exact tests (P<0.05). RESULTS: Mean age of women in groups A and B were 35.6±4.0 and 34.9±3.9 respectively (NS). A total of 89 matured oocytes were collected in group A and 127 in group B. All of them were submitted to 1stPB biopsy. Some oocytes were in Telophase I stage and therefore were excluded from the genetic test [8% and 11% in group A and B, respectively (NS)]. Eighty-one and 113 1stPBs from group 1 and 2 were sent for genetic analysis. The proportions of unaffected oocytes were similar in groups 1 and 2 [20% and 24% respectively (NS)]. In group 2, two cycles did not result in any genetically normal oocytes. In group 1 and 2, 12 and 25 oocytes were inseminated yielding a fertilization rate of 67% and 72% respectively (NS). There was not a statistically significant difference in the percentages of developing embryos (Grades A, B and C) that were 37.5%, 12.5% and 50% (group 1); and 52%, 33% and 14% (group 2), respectively. In group 1, 8 embryos were transferred in 6 ET cycles resulting in one healthy baby born. In group 2, 18 embryos were transferred in 10 ET cycles yielding 2 healthy clinical pregnancies that are still ongoing. CONCLUSIONS: The results of this study demonstrate that removing the 1stPB is similarly efficient by the two approaches applied. Both techniques resulted in a reasonably good fertilization and embryo development. However, few pregnancies were achieved, independently of the type of PB removing technique. It is possible that the manipulation during the PB biopsy has an impact on oocyte physiology or subcellular structure that later would influence the derived embryo viability. Alternatively, a longer than ideal time of incubation between oocyte collection and insemination may affect the oocyte quality and competence. Further studies are required to evaluate these possibilities with the intent of improving clinical outcome.