Lsoelectric focusing phenotyping and denaturing gradient gel electrophoresis genotyping: a comparison of two methods in detection of alpha-1-antitrypsin variants
Само за регистроване кориснике
2008
Аутори
Ljujić, MilaTopić, Aleksandra
Divac Rankov, Aleksandra
Nikolić, Aleksandra
Petrović-Stanojević, Nataša
Surlan, Mirjana
Mitić-Milikić, Maria
Radojković, Dragica
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
Laboratory diagnosis of alpha-1-antitrypsin (AAT) deficiency is routinely performed by phenotyping methods, which include measurement of serum alpha-1-antitrypsin concentration and isoelectric focusing (IEF). Several DNA-based methods are also used for AAT deficiency testing, but they still have not become part of routine diagnostics. The aim of the study was to identify AAT variants using 2 different methods, isoelectric focusing and denaturing gradient gel electrophoresis (DGGE), and to compare obtained results as well as practical application of these 2 methods. The study has encompassed 27 emphysema patients. In all patients, AAT phenotypization was conducted using IEF, whereas genotypization was performed by DGGE. Variations detected by DGGE were characterized by DNA sequencing. Mutations in the AAT gene were detected in 6 patients. Three patients were homozygous for the Z allele, whereas I patient was heterozygous. In 2 patients, novel AAT variants, G320R and V321F, were detected.... When results obtained by IEF and DGGE were compared, it was observed that IEF results were inconclusive or misinterpreted in 5 cases (18.5%). Both methods proved to be reliable for detection of the Z alleles, whereas discrepancy existed for M4 allele and rare variants. Therefore, the optimal strategy for diagnostics of AAT deficiency should encompass detection of the most common AAT variants by IEF and screening for the less common variants by DGGE in combination with sequencing.
Извор:
Translational Research, 2008, 151, 5, 255-259Издавач:
- Elsevier Science Inc, New York
Финансирање / пројекти:
- Структурални елементи генома у модулацији фенотипа (RS-MESTD-MPN2006-2010-143051)
DOI: 10.1016/j.trsl.2008.02.002
ISSN: 1931-5244
PubMed: 18433707
WoS: 000255495700004
Scopus: 2-s2.0-42349108135
Институција/група
Institut za molekularnu genetiku i genetičko inženjerstvoTY - JOUR AU - Ljujić, Mila AU - Topić, Aleksandra AU - Divac Rankov, Aleksandra AU - Nikolić, Aleksandra AU - Petrović-Stanojević, Nataša AU - Surlan, Mirjana AU - Mitić-Milikić, Maria AU - Radojković, Dragica PY - 2008 UR - https://imagine.imgge.bg.ac.rs/handle/123456789/340 AB - Laboratory diagnosis of alpha-1-antitrypsin (AAT) deficiency is routinely performed by phenotyping methods, which include measurement of serum alpha-1-antitrypsin concentration and isoelectric focusing (IEF). Several DNA-based methods are also used for AAT deficiency testing, but they still have not become part of routine diagnostics. The aim of the study was to identify AAT variants using 2 different methods, isoelectric focusing and denaturing gradient gel electrophoresis (DGGE), and to compare obtained results as well as practical application of these 2 methods. The study has encompassed 27 emphysema patients. In all patients, AAT phenotypization was conducted using IEF, whereas genotypization was performed by DGGE. Variations detected by DGGE were characterized by DNA sequencing. Mutations in the AAT gene were detected in 6 patients. Three patients were homozygous for the Z allele, whereas I patient was heterozygous. In 2 patients, novel AAT variants, G320R and V321F, were detected. When results obtained by IEF and DGGE were compared, it was observed that IEF results were inconclusive or misinterpreted in 5 cases (18.5%). Both methods proved to be reliable for detection of the Z alleles, whereas discrepancy existed for M4 allele and rare variants. Therefore, the optimal strategy for diagnostics of AAT deficiency should encompass detection of the most common AAT variants by IEF and screening for the less common variants by DGGE in combination with sequencing. PB - Elsevier Science Inc, New York T2 - Translational Research T1 - Lsoelectric focusing phenotyping and denaturing gradient gel electrophoresis genotyping: a comparison of two methods in detection of alpha-1-antitrypsin variants EP - 259 IS - 5 SP - 255 VL - 151 DO - 10.1016/j.trsl.2008.02.002 ER -
@article{ author = "Ljujić, Mila and Topić, Aleksandra and Divac Rankov, Aleksandra and Nikolić, Aleksandra and Petrović-Stanojević, Nataša and Surlan, Mirjana and Mitić-Milikić, Maria and Radojković, Dragica", year = "2008", abstract = "Laboratory diagnosis of alpha-1-antitrypsin (AAT) deficiency is routinely performed by phenotyping methods, which include measurement of serum alpha-1-antitrypsin concentration and isoelectric focusing (IEF). Several DNA-based methods are also used for AAT deficiency testing, but they still have not become part of routine diagnostics. The aim of the study was to identify AAT variants using 2 different methods, isoelectric focusing and denaturing gradient gel electrophoresis (DGGE), and to compare obtained results as well as practical application of these 2 methods. The study has encompassed 27 emphysema patients. In all patients, AAT phenotypization was conducted using IEF, whereas genotypization was performed by DGGE. Variations detected by DGGE were characterized by DNA sequencing. Mutations in the AAT gene were detected in 6 patients. Three patients were homozygous for the Z allele, whereas I patient was heterozygous. In 2 patients, novel AAT variants, G320R and V321F, were detected. When results obtained by IEF and DGGE were compared, it was observed that IEF results were inconclusive or misinterpreted in 5 cases (18.5%). Both methods proved to be reliable for detection of the Z alleles, whereas discrepancy existed for M4 allele and rare variants. Therefore, the optimal strategy for diagnostics of AAT deficiency should encompass detection of the most common AAT variants by IEF and screening for the less common variants by DGGE in combination with sequencing.", publisher = "Elsevier Science Inc, New York", journal = "Translational Research", title = "Lsoelectric focusing phenotyping and denaturing gradient gel electrophoresis genotyping: a comparison of two methods in detection of alpha-1-antitrypsin variants", pages = "259-255", number = "5", volume = "151", doi = "10.1016/j.trsl.2008.02.002" }
Ljujić, M., Topić, A., Divac Rankov, A., Nikolić, A., Petrović-Stanojević, N., Surlan, M., Mitić-Milikić, M.,& Radojković, D.. (2008). Lsoelectric focusing phenotyping and denaturing gradient gel electrophoresis genotyping: a comparison of two methods in detection of alpha-1-antitrypsin variants. in Translational Research Elsevier Science Inc, New York., 151(5), 255-259. https://doi.org/10.1016/j.trsl.2008.02.002
Ljujić M, Topić A, Divac Rankov A, Nikolić A, Petrović-Stanojević N, Surlan M, Mitić-Milikić M, Radojković D. Lsoelectric focusing phenotyping and denaturing gradient gel electrophoresis genotyping: a comparison of two methods in detection of alpha-1-antitrypsin variants. in Translational Research. 2008;151(5):255-259. doi:10.1016/j.trsl.2008.02.002 .
Ljujić, Mila, Topić, Aleksandra, Divac Rankov, Aleksandra, Nikolić, Aleksandra, Petrović-Stanojević, Nataša, Surlan, Mirjana, Mitić-Milikić, Maria, Radojković, Dragica, "Lsoelectric focusing phenotyping and denaturing gradient gel electrophoresis genotyping: a comparison of two methods in detection of alpha-1-antitrypsin variants" in Translational Research, 151, no. 5 (2008):255-259, https://doi.org/10.1016/j.trsl.2008.02.002 . .