Development of a multiplex selected reaction monitoring assay for quantification of biochemical markers of down syndrome in amniotic fluid samples.
Down syndrome (DS) is one of the most common chromosomal abnormalities affecting about 1 of every 700 fetuses. Current screening strategies have detection rates of 90-95% at a 5% false positive rate. The aim of this study was to discover new biomarkers of DS in amniotic fluid by using a multiplex selected reaction monitoring assay. Nine proteins were analyzed: CEL, CPA1, MUC13, CLCA1, MUC5AC, PLUNC, and HAPLN1, and CGB as positive control and serotransferrin as negative control. One proteotypic peptide for each protein was selected, and internal heavy isotope-labeled peptide standards were spiked into the samples. Fifty-four samples from pregnant women carrying normal (n = 37) or DS-affected (n = 17) fetuses were analyzed. The median protein concentrations for DS and normal samples, respectively, were as follows: 20 and 49 ng/mL (p < 0.01) for CEL; 3.7 and 14 ng/mL (p < 0.001) for CPA1; 80 and 263 ng/mL (p < 0.001) for MUC13; 46 and 135 ng/mL (p < 0.001) for CLCA1; 0.65 and 0.93 μg/mL (p < 0.05) for MUC5AC; 61 and 73 ng/mL (p > 0.05) for PLUNC; 144 and 86 ng/mL (p < 0.01) for HAPLN1; 0.89 and 0.54 μg/mL (p = 0.05) for CGB; 91 and 87 μg/mL (p > 0.05) for serotransferrin. Statistically significant differences were found in six out of the seven candidate proteins analyzed, reflecting a different regulation in DS.
Samuel Lunenfeld Research Institute, Mount Sinai Hospital, Toronto, Ontario, Canada.
Eduardo
E
Martínez-Morillo
Chan-Kyung J
CK
Cho
Andrei P
AP
Drabovich
Julie L V
JL
Shaw
Antoninus
A
Soosaipillai
Eleftherios P
EP
Diamandis
D000649:Amniocentesis; D000653:Amniotic Fluid / Q000378:metabolism*; D019540:Area Under Curve; D015415:Biomarkers / Q000378:metabolism; D043422:Carboxypeptidases A / Q000378:metabolism; D016022:Case-Control Studies; D018118:Chloride Channels / Q000378:metabolism; D004314:Down Syndrome / Q000175:diagnosis* / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005260:Female; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D008049:Lipase / Q000378:metabolism; D055271:Mucin 5AC / Q000378:metabolism; D009077:Mucins / Q000378:metabolism; D010750:Phosphoproteins / Q000378:metabolism; D011247:Pregnancy; D011509:Proteoglycans / Q000378:metabolism; D012372:ROC Curve; D053719:Tandem Mass Spectrometry; D000099249:Proteoglycan Link Protein
D015415:Biomarkers; D043422:Carboxypeptidases A; D018118:Chloride Channels; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; D008049:Lipase; D055271:Mucin 5AC; D009077:Mucins; D010750:Phosphoproteins; D011509:Proteoglycans; D000099249:Proteoglycan Link Protein; C485599:BPIFA1 protein, human; C492109:CLCA1 protein, human; C429552:MUC13 protein, human; C524678:MUC5AC protein, human; C054621:CEL protein, human