title: Molecular Diagnosis of α⁰-Thalassemia Through Urine DNA: A Novel DNA Source to Facilitate Prevention Programs in Remote Geographical Areas.
abstract: We assessed whether urinary DNA sediment was a feasible sample type for the molecular diagnosis of α-thalassemia (α-thal) mutations. Urine samples (5-10 mL) were collected from 218 male and female volunteers. The cells were centrifuged, and DNA was isolated according to the protocol of a commercial DNA isolation kit. Detection of the α(0)-thal [Southeast Asian (- -(SEA)) and - -(THAI)] deletions was performed using quantitative real-time polymerase chain reaction (q-PCR), in addition to conventional gap-PCR. The results revealed that DNA extracted from urinary sediment presented an average DNA content of 11.2 ± 5.5 ng/µL, and the 260/280 ratio indicative of DNA purity, was 1.2 ± 0.2. The overall q-PCR threshold cycle was 31.2 ± 2.3. The melting temperature for the - -(SEA) deletion was 87.3 ± 0.1 °C, while that of the wild type sequence was 92.5 ± 0.2 °C. There were 16 (7.3%) α(0)-thal SEA genotypes detected. These results were in agreement with those of the conventional gap-PCR and blood DNA analyses. Thus, DNA from urinary sediment can be efficiently used for the molecular diagnosis of α(0)-thal mutations. This approach allows for rapid diagnosis, is non invasive, and could be useful for preventing Hb Bart's (γ4) hydrops fetalis syndrome.
authors.affiliation: Discipline of Biology, School of Science, University of Phayao , Phayao , Thailand.
authors.forename: Narutchala
authors.initials: N
authors.lastname: Suwannakhon
authors.forename: Teerapat
authors.initials: T
authors.lastname: Seeratanachot
authors.forename: Khwanruedee
authors.initials: K
authors.lastname: Mahingsa
authors.forename: Torpong
authors.initials: T
authors.lastname: Sanguansermsri
mesh_terms: D000328:Adult; D004247:DNA / Q000652:urine*; D005260:Female; D006455:Hemoglobins, Abnormal / Q000235:genetics; D006801:Humans; D015160:Hydrops Fetalis / Q000175:diagnosis / Q000235:genetics / Q000517:prevention & control; D008297:Male; D060888:Real-Time Polymerase Chain Reaction; D055815:Young Adult; D055542:alpha-Globins / Q000235:genetics*; D017085:alpha-Thalassemia / Q000175:diagnosis* / Q000235:genetics* / Q000517:prevention & control
chemical_list: D006455:Hemoglobins, Abnormal; D055542:alpha-Globins; D004247:DNA; C030448:hemoglobin Bart's
keywords: molecular diagnosis; mutation; urine DNA; α0-Thalassemia (α0-thal)