QF-PCR and MLPA: a reliable molecular system to detect chromosomal alterations in miscarriages

Area of Science:

• Genetics
• Reproductive Biology
• Molecular Diagnostics

Background:

• Miscarriage analysis traditionally relies on cytogenetic testing, which can be time-consuming and may have limitations in detecting specific chromosomal abnormalities.
• Accurate genetic analysis of miscarriage products is crucial for understanding reproductive failure and providing genetic counseling.

Purpose of the Study:

• To evaluate the efficacy of a combined quantitative fluorescent-polymerase chain reaction (QF-PCR) and multiplex ligation-dependent probe amplification (MLPA) system for detecting chromosome alterations in miscarriage samples.
• To establish this combined QF-PCR/MLPA approach as a viable alternative to conventional cytogenetic methods.

Main Methods:

• A total of 264 miscarriage samples were analyzed between 2011 and 2015 using the combined QF-PCR/MLPA system.
• QF-PCR was employed for initial screening of chromosomes 13, 18, 21, X, and Y. Maternal DNA analysis was performed to confirm fetal origin.
• MLPA analysis was subsequently conducted on subtelomeric regions for samples of confirmed fetal origin.

Main Results:

• Of 264 samples, 229 were of fetal origin, with 46.3% showing pathological results.
• Pathological findings included autosomal aneuploidies (74.4%), triploidies (10.4%), sex chromosomal aneuploidies (8.5%), structural alterations (3.7%), and multiple aneuploidies (2.7%).
• QF-PCR yielded results for all samples, while MLPA provided unambiguous results in approximately 90% of cases.

Conclusions:

• The combined QF-PCR/MLPA system is highly effective in detecting a wide range of chromosomal abnormalities, including aneuploidies and triploidies.
• This approach accurately identifies structural unbalanced alterations in subtelomeric regions.
• The QF-PCR/MLPA system presents a robust and efficient alternative for miscarriage genetic analysis.