MLPA-based evidence for sequence gain: pitfalls in confirmation and necessity for exclusion of false positives.
Multiplex ligation-dependent probe amplification (MLPA) has become a standard method for identifying copy number mutations in diagnostic and research settings. The occurrence of false-positive deletion findings and the underlying causes are well recognized, whereas false-positive duplication/amplification findings have not been appreciated so far. We here present three pertinent cases which were only identified on extended, nonstandard secondary analyses. We also offer and experimentally validate a potential explanation. Our findings imply that MLPA data indicating gain of genomic sequence require validation on an independent sample or by an independent method.
Department of Clinical Chemistry and Laboratory Diagnostics, University Hospital Jena, Jena, Germany.
Rita-Eva
RE
Varga
Rizwan
R
Mumtaz
Amir
A
Jahic
Galina E
GE
Rudenskaya
Elena
E
Sánchez-Ferrero
Michaela
M
Auer-Grumbach
Christian A
CA
Hübner
Christian
C
Beetz
D005189:False Positive Reactions; D018628:Gene Dosage; D060885:Multiplex Polymerase Chain Reaction / Q000379:methods*; D017384:Sequence Deletion