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OBJECTIVE: This study aims to investigate the impact of refrigeration time and blood volume on the success rate of peripheral blood chromosomal analysis using response surface methodology (RSM). METHODS: Peripheral blood samples from 30 volunteers were subjected to chromosomal analysis under different refrigeration duration periods (≤7 days, 8 days, 9 days, 10 days, 11 days, 12 days, 13 days, 14 days) along with different blood volumes (0.2 mL, 0.3 mL, 0.4 mL, 0.5 mL, 0.6 mL, 0.7 mL, and 0.8 mL). The effects of refrigeration time and blood volume on the success rate of peripheral blood chromosomal analysis were determined using the Chi-square test for trend, followed with Spearman's rank correlation coefficient, and RSM analysis to identify the optimal combination of refrigeration time and blood volume. RESULTS: The refrigeration time within 10 days had a minor impact on the success rate, while refrigeration time more than 11 days significantly decreased the success rate. An increase in blood volume slightly improved the success rate. The success rate showed both linear and nonlinear changes with refrigeration time, while the effect of blood volume was primarily linear. The highest success rate was observed at a refrigeration time of ≤7 days and a blood volume of 0.8 mL. The interaction between refrigeration time and blood volume had a significant impact on the success rate. CONCLUSION: It is recommended to keep the refrigeration time of blood samples within 7 days and control the blood volume at 0.8 mL to maximize the success rate of chromosomal analysis.
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BACKGROUND: Using combined fluorescence in situ hybridization (FISH) and high-throughput whole-genome sequencing (WGS) molecular cytogenetic technology, we aim to analyze the junction breakpoints of complex chromosome rearrangements (CCR) that were difficult to identify by conventional karyotyping analysis and further characterize the genetic causes of recurrent spontaneous abortion. RESULTS: By leveraging a combination of current molecular techniques, including chromosome karyotype analysis, FISH, and WGS, we comprehensively characterized the extremely complex chromosomal abnormalities in this patient with recurrent spontaneous abortions. Here, we demonstrated that combining these current established molecular techniques is an effective and efficient workflow to identify the structural abnormalities of complex chromosomes and locate the rearrangement of DNA fragments. CONCLUSIONS: In conclusion, leveraging results from multiple molecular and cytogenetic techniques can provide the most comprehensive genetic analysis for genetic etiology research, diagnosis, and genetic counseling for patients with recurrent spontaneous abortion and embryonic abortion.
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This study aimed to analyze the clinical phenotype of chromosome 9p deletion or duplication and its relationship with karyotype. A patient, female, aged 6 months, visited the hospital due to motor developmental delay. Karyotype analysis identified abnormalities of chromosome 9 short arm, and high-throughput sequencing found 9p24.3-9p23 deletion and 9p23-9p13.1 duplication. Her parents had a normal karyotype. Karyotype analysis combined with high-throughput sequencing is of great significance for improving the efficiency of etiological diagnosis in children with motor developmental delay or multiple congenital deformities and mental retardation.
