MQS-PCR: a simple and quick multiplex PCR molecular method for human chromosomal sex determination in DNA samples
Quantitative PCR puts great demands on DNA quality and relies on a comparison of fragment amplification between two chromosomes using different primers. The use of a single primer pair capable of reliable relative comparative amplification would be a great advantage. Using this approach, we developed a rapid, high-throughput, semi-automated and cost-efficient multiplexed method for molecular determination of chromosomal sex called MQS-PCR – multiplex quantitative sexing PCR. DNA sequences located on different chromosomes and differing in length can be amplified and fluorescently labelled with a common fluorescent primer and conveniently separated and detected using capillary electrophoresis. In this method, the intensity of amplification of each of the fragments is compared to determine DNA dosage. MQS-PCR achieves 100% analytical sensitivity and specificity in detecting normal and abnormal sex chromosomal complements (45,X, 46,X,i(X)(q10), 46,X,i(X)(p10), 46,X,X(p-), 46,X,X(q-), 47,XXX, 47,XXY and 47,XYY). It is a reliable, low cost, and rapid detection method for the determination of chromosomal sex and sex chromosomal abnormalities in human samples.