RESUMO
Background: The purpose of the study was to investigate the frequencies and types of Y chromosome microdeletions in infertile men and to analyze the relationship between the levels of reproductive hormones and Y microdeletions
Methods: A total of 1,226 infertile men were screened for Y chromosome microdeletions using multiplex PCR assay. Karyotype analysis was performed on peripheral blood lymphocytes with standard G-banding. Serum reproductive hormone levels were measured
Results: Out of 1,226 infertile patients, 134 [10.93%] had Y microdeletions. One hundred seven of 765 [13.99%] non-obstructive azoospermic patients and 27 of 133 [20.30%] severe oligozoospermic patients had Y microdeletions. Among the 134 infertile men with Y microdeletions, the most frequent microdeletions were detected in the AZFc region, followed by AZFbc, AZFb, AZFa, AZFabc[Yq], Yp[SRY]+Yq, and partial AZFc regions. Karyotype analysis was available for 130 of the 134 patients with Y microdeletions. Of them, 36 [27.69%] patients had sex chromosomal abnormalities. Levels of FSH and LH in patients with AZFc microdeletion were significantly lower, while those in patients with Yp[SRY]+Yq were significantly higher than in patients without Y microdeletions. Level of testosterone in patients with AZFabc[Yq] or Yp[SRY]+Yq was significantly lower than that in patients without Y microdeletions. However, there was no significant difference in the levels of reproductive hormones between all patients with and without Y microdeletions
Conclusion: These results highlight the need for Y chromosome microdeletion screening for correct diagnosis of male infertility. Obtaining reliable genetic information for assisted reproductive techniques can prevent unnecessary treatment and vertical transmission of genetic defects to offspring
RESUMO
PURPOSE: We applied a simplified method using polymerase chain reaction (PCR)-based enzymatic digestion for the detection of epidermal growth factor receptor (EGFR) mutation. MATERIALS AND METHODS: We selected 74 samples of adenocarcinoma of the lung with EGFR exons 19 and 21 that had been previously sequenced. We designed PCR primers and chose a DNA restriction enzyme. Seventy four additional lung cancer samples were tested as a test set. For test sets, the PCR-based method was performed first, followed by validation of the result by DNA sequencing. RESULTS: In the first sample group, we found 15 (20.3%) mutations in exon 19, and 9 (12.2%) mutations in exon 21 using the sequencing method. By using the PCR-based method, we were able to identify all of the mutated samples detected by the sequencing method. The PCR-based method also detected mutations in exon 19 in three additional samples and in exon 21 in one additional sample. In the second sample group, by performing the PCR-based method, we found 10 (13.5%) and 7 (9.5%) mutations in exons 19 and 21, respectively. Additional mutations in exon 19 were identified in 2 samples by the sequencing method. However, the sequencing method failed to identify a mutation in exon 21 in one sample. CONCLUSION: The sensitivity of the PCR-based enzymatic digestion method seems to be comparable to that of the traditional sequencing method for detecting EGFR mutations. Our method can be widely used as a screening test to select patients who may benefit from EGFR targeted therapy.