RESUMO

Liquid-based cytology (LBC) has been used as a diagnostic tool for cervical cancer for years and is now being adopted for other gynecological cancers. LBC represents an important challenge to ensure that the process yields representative biospecimens for quality control (QC) of diagnostic procedures. In this study, we compare QC parameters (integrity, yield and purity, and polymerase chain reaction [PCR] amplification) of DNA isolated from LBC (N = 296) using two different nucleic acid isolation methods, manual (n = 233) or automated (n = 63). We also evaluated two different types of cytological brushes for sampling from the cervix. Our results suggest that manual isolation (yield 22.81 ± 1.92 µg) resulted in increased DNA recovery when compared with automated isolation (yield 9.96 ± 1.11 µg) from LBC samples, with a p-value of <0.0003. We estimated that 98% (53/54) of the samples preserved the integrity of DNA and were suitable for standard molecular biology analyses. The ß-globin gene was amplified in 100% (296/296) of the DNA samples by endpoint PCR. We found no significant difference between the performance of the cytological brushes (p value of <0.6711) in a general overview. However, when looking at the results from using each brush individually, the manual isolation method was statistically superior to the automated method. Our work illustrates the impact of good QC of preanalytic conditions, which will be important for the application of LBC for developing early detection methods for gynecological cancers.

Assuntos

DNA de Neoplasias/isolamento & purificação , Manejo de Espécimes/métodos , Neoplasias do Colo do Útero/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Bancos de Espécimes Biológicos , Detecção Precoce de Câncer , Feminino , Humanos , Biópsia Líquida , Neoplasias do Colo do Útero/genética , Adulto Jovem

RESUMO

BACKGROUND: One determining factor of a successful in vitro fertilization (IVF) cycle is embryo quality. The aim of the present study was to evaluate associations of embryo quality and reserve markers like age, FSH and AMH. MATERIALS AND METHODS: In this prospective study, 120 infertile women, aged 21-44 years, undergoing routine exploration during an unstimulated cycle preceding assisted reproductive technology (ART) at our center were studied prospectively, from February 2011 to December 2014. Descriptive parameters and patient characteristics were reported as mean (SD) or median (range) depending on the distribution. Student's t test was performed for continuous variables, Wilcoxon and Pearson's Test were used for not distributed variables and Fisher's Test was performed for categorical variables. P<0.05 was considered statistically significant. RESULTS: Overall, at the time of investigation, patients had a mean age of 33.03 ± 4.15 years old. On cycle day three, serum anti-Mullerian hormone (AMH) level was 3.50 ± 1.54 ng/mL, serum follicle-stimulating hormone (FSH) level was 6.29 ± 1.53 mUI/ mL, at baseline, women had 16.57 ± 7.0 antral follicles. The mean of collected oocytes was 11.80 ± 5.25, embryo I+II was 2.46 ± 2.11. A greater number of embryos I+II was observed in young patients. By evaluating 120 patients, a significant relationship was observed between age and FSH (r=0.24, P=0.01), age with AMH (r=-0.22, P=0.02), age with collected oocytes (r=-0.23, P=0.03) and age with embryo I+II (r=-0.22, P=0.03). A significant relationship was also observed between antral follicle count (AFC) and AMH (r=0.29, P=0.01), AFC and the number of transferred embryo (r=-0.18, P=0.03), AFC and total dose of the drugs (r=-0.23, P=0.03). Significant relationship of FSH with total dose of drugs (r=0.19, P=0.02) was also observed. In addition, we determined significant relationships between AMH and the number of collected oocytes (r=0.38, P=0.01), AMH and the number of metaphase II oocytes (r= 0.35, P=0.01), AMH and the number of embryo (r=0.19, P=0.04) as well as AMH and total dose of the drugs (r=-0.25, P=0.01). CONCLUSION: Commonly used clinical markers of ovarian reserve are reflection of the ovarian reserve, while the outcome measurements of ART and age are the best predictors of embryo quality.