Combination of QF‐PCR and aCGH is an efficient diagnostic strategy for the detection of chromosome aberrations in recurrent miscarriage

Abstract

BackgroundOur aim was to conduct a comprehensive genetic evaluation using the combination of QF‐PCR (quantitative fluorescence polymerase chain reaction) and aCGH (array comparative genomic hybridization) for the detection of the frequency and type of chromosome aberrations in recurrent miscarriage (RM) in the clinical setting.MethodsThis retrospective study was conducted on 73 first‐trimester products of conception (POC) between September 2014 and February 2017. The POCs were collected from 73 women with at least one previous miscarriage and analyzed for chromosomal anomalies using QF‐PCR and aCGH as part of the routine clinical evaluation.ResultsChromosome aberrations were detected in 52/73 POCs (71.2%), of which 41 (56.2%) were identified by QF‐PCR and an additional 11 (15.1%) by aCGH. Numerical aberrations constituted 92.3% of abnormalities, with trisomies as the most common subtype (72.9%). Causative structural aberrations were found in three samples (5.8%). The frequency of chromosome aberrations was not dependent on the number of previous miscarriages, whereas it significantly increased with advanced maternal age.ConclusionOur results confirm that chromosome aberrations are the most common cause of RM and that QF‐PCR and aCGH combination should be included in the routine genetic analysis of POCs of couples with miscarriage.

Usefulness of combined NGS and QF‐PCR analysis for product of conception karyotyping

“… 6 , 7 , 8 Recently, SNP microarray, next‐generation sequencing (NGS), quantitative fluorescence PCR (QF‐PCR), and multiplex ligation‐dependent probe amplification (MLPA), none of which require cell culture, have been reported as alternative approaches to the cytogenomic analysis of a POC. 9 , 10 , 11 , 12 , 13 , 34 …”

“…[6][7][8] Recently, SNP microarray, next-generation sequencing (NGS), quantitative fluorescence PCR (QF-PCR), and multiplex ligation-dependent probe amplification (MLPA), none of which require cell culture, have been reported as alternative approaches to the cytogenomic analysis of a POC. [9][10][11][12][13]34 NGS is a powerful tool that can allow both qualitative and quantitative analyses to be performed simultaneously. Currently, NGS-based chromosome analysis is mainly used in the preimplantation genetic testing for aneuploidy and structural rearrangements (PGT-A and PGT-SR), using the amplified products of the whole genome from the biopsied trophectoderm cells of a 5day embryo.…”

“…The G‐banding of POC samples has practical limitations related to the need for cell culturing which can often fail due to fetal demise or macerated tissue, the preferential growth of maternal decidua cells, and the emergence of artifacts 6–8 . Recently, SNP microarray, next‐generation sequencing (NGS), quantitative fluorescence PCR (QF‐PCR), and multiplex ligation‐dependent probe amplification (MLPA), none of which require cell culture, have been reported as alternative approaches to the cytogenomic analysis of a POC 9–13,34 …”

Diagnostic accuracy and value of chromosomal microarray analysis for chromosomal abnormalities in prenatal detection

“…At present, molecular cytogenetic techniques including multiplex ligation-dependent probe amplification, quantitative fluorescence polymerase chain reaction, and fluorescence in situ hybridization, are gradually applied to detect submicroscopic chromosomal aberrations in clinical practices. [ 7 , 8 ] However, these methods are not feasible to detect all possible chromosome deletion and duplication.…”

Diagnostic accuracy and value of chromosomal microarray analysis for chromosomal abnormalities in prenatal detection: a prospective clinical study

“…However, karyotyping has considerable limitations, especially for the lack of detection of many unbalanced structural abnormalities from submicroscopic chromosomal aberrations. In recent years, molecular cytogenetic methods, including multiplex ligationdependent probe ampli cation (MLPA), quantitative uorescence polymerase chain reaction (QF-PCR), and uorescence in situ hybridization (FISH), are gradually applied to evaluate submicroscopic chromosomal aberrations [7,8]. However, these techniques are not feasible to detect all possible chromosome deletion and duplication.…”