Περίληψη:
Background: The detection and diagnosis of beta-thalassaemia for
populations with molecular heterogeneity, or diverse ethnic groups, has
increased the need for the development of an array high-throughput
diagnostic tool that can deliver large scale genetic detection. We
report on the update and validation of the ThalassoChip, a
beta-thalassaemia genetic diagnostic tool which is based on arrayed
primer extension (APEX) technology.
Methods: ThalassoChip slides with new and redesigned probes were
prepared for testing the microarray. Six hundred and sixty DNA samples
collected from eight Mediterranean countries were used for
standardisation, optimisation and validation of the ThalassoChip. The
beta-globin gene region was amplified by PCR, the products were
hybridised to the probes after fragmentation and the APEX reaction
followed.
Results: The ThalassoChip was updated with new probes and now has the
ability to detect 57 beta-globin gene mutations and three single
nucleotide polymorphisms (SNPs) in a single test. The ThalassoChip as
well as the PCR and APEX reactions were standardised and optimised using
500 DNA samples that were previously genotyped using conventional
diagnostic techniques. Some probes were redesigned in order to improve
the specificity and sensitivity of the test. Validation of the
ThalassoChip performed using 160 samples analysed in blinded fashion
showed no error.
Conclusions: The updated version of the ThalassoChip is versatile,
robust, cost-effective and easily adaptable, but most notably can
provide comprehensive genetic diagnosis for beta-thalassaemia and other
haemoglobinopathies. Clin Chem Lab Med 2010; 48:1713-8.
Συγγραφείς:
Shammas, Christos
Papasavva, Thessalia
Felekis, Xenia and
Christophorou, Christos
Roomere, Hanno
Synodinos, Jan Traeger
and Kanavakis, Emmanuel
El-Khateeb, Mohammed
Hamamy, Hanan and
Mahmoud, Tamara
Shboul, Mohammad
El Beshlawy, Amal
Filon,
Dvora
Hussein, Ibtessam R.
Galanello, Renzo
Romeo, Giovanni
and Kleanthous, Marina
