Identification of the four most common beta-globin gene mutations in Greek beta-thalassemic patients and carriers by PCR-SSCP: advantages and limitations of the method
Φόρτωση...
Ημερομηνία
Συγγραφείς
Kakavas, K. V.
Noulas, A.
Chalkias, C.
Hadjichristodoulou, C.
Georgiou, I.
Georgatsou, E.
Bonanou, S.
Τίτλος Εφημερίδας
Περιοδικό ISSN
Τίτλος τόμου
Εκδότης
Περίληψη
Τύπος
Είδος δημοσίευσης σε συνέδριο
Είδος περιοδικού
peer-reviewed
Είδος εκπαιδευτικού υλικού
Όνομα συνεδρίου
Όνομα περιοδικού
J Clin Lab Anal
Όνομα βιβλίου
Σειρά βιβλίου
Έκδοση βιβλίου
Συμπληρωματικός/δευτερεύων τίτλος
Περιγραφή
In the present study we investigated whether the single-strand conformational polymorphism (SSCP) method could be employed to identify (rather than simply detect) the four most common beta-globin gene mutations in the Greek population: IVS-I-110, Cd39, IVS-I-1, and IVS-I-6. Using DNA from 50 beta-thalassemic patients and carriers, we amplified by PCR the appropriate 238-bp region of the human beta-globin gene, analyzed the reaction products by nondenaturing polyacrylamide gel electrophoresis, and visualized the bands by silver staining. Single-stranded DNA (ssDNA) fragments showed a reproducible pattern of bands that was characteristic of the mutations present. With the use of control samples containing six of the 10 possible combinations of the four most common beta-globin gene mutations, we were able to predict the mutations present in a quarter of the patients studied. Our predictions were confirmed independently by the amplification refractory mutation system (ARMS) method. We conclude that this non-radioactive PCR-SSCP method can be used to reliably identify mutations in patients, provided that suitable controls are available. Moreover, the method is easy to apply to the identification of mutations in carriers, which makes it particularly useful for population screening.
Περιγραφή
Λέξεις-κλειδιά
Electrophoresis, Polyacrylamide Gel, Genotype, Globins/*genetics, *Heterozygote, Humans, Point Mutation, Polymerase Chain Reaction/*methods, *Polymorphism, Single-Stranded Conformational, beta-Thalassemia/*genetics
Θεματική κατηγορία
Παραπομπή
Σύνδεσμος
http://www.ncbi.nlm.nih.gov/pubmed/16470532
http://onlinelibrary.wiley.com/store/10.1002/jcla.20091/asset/20091_ftp.pdf?v=1&t=h0j4onk9&s=5672d7115ff6274bc2d933f24dea328183993a8d
http://onlinelibrary.wiley.com/store/10.1002/jcla.20091/asset/20091_ftp.pdf?v=1&t=h0j4onk9&s=5672d7115ff6274bc2d933f24dea328183993a8d
Γλώσσα
en
Εκδίδον τμήμα/τομέας
Όνομα επιβλέποντος
Εξεταστική επιτροπή
Γενική Περιγραφή / Σχόλια
Ίδρυμα και Σχολή/Τμήμα του υποβάλλοντος
Πανεπιστήμιο Ιωαννίνων. Σχολή Επιστημών Υγείας. Τμήμα Ιατρικής