Identification of the four most common β‐globin gene mutations in Greek β‐thalassemic patients and carriers by PCR‐SSCP: advantages and limitations of the method

Kakavas, Konstantinos; Noulas, A.; Chalkias, Christos; Hadjichristodoulou, Christos; Georgiou, Ioannis; Georgatsou, Eleni; Bonanou, Sophia

doi:10.1002/jcla.20091

Cited by 5 publications

(3 citation statements)

References 20 publications

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“…Other working groups may initially prefer SSCP for the mutations screening and subsequently refinement under alternate condition or in combination with heterodublex analysis to increase accuracy. The SSCP method can also be employed for the identification and not simply the detection of the mutations [41] (Table 3).…”

Section: Simultaneous Performance Of Ha and Sscpmentioning

confidence: 99%

PCR–SSCP: A Method for the Molecular Analysis of Genetic Diseases

et al. 2007

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Expression of certain transgenes from an adenovirus vector can be deleterious to its own replication. This can result in the inhibition of virus rescue, reduced viral yields, or, in the worst case, make it impossible to construct a vector expressing the inhibiting transgene product. A gene regulation system based on the tet operon was used to allow the rescue and efficient growth of adenovectors that express transgenes to high levels. A key advantage to this system is that repression of transgene expression is mediated by the packaging cell line, thus, expression of regulatory products from the adenovector are not required. This provides a simple, broadly applicable system wherein transgene repression is constitutive during vector rescue and growth and there is no effect on adenovector-mediated expression of gene products in transduced cells. Several high-level expression vectors based on first- and second-generation adenovectors were rescued and produced to high titer that otherwise could not be grown. Yields of adenovectors expressing inhibitory transgene products were increased, and the overgrowth of cultures by adenovectors with nonfunctional expression cassettes was prevented. The gene regulation system is a significant advancement for the development of adenovirus vectors for vaccine and other gene transfer applications.

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Section: Simultaneous Performance Of Ha and Sscpmentioning

confidence: 99%

PCR–SSCP: A Method for the Molecular Analysis of Genetic Diseases

et al. 2007

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“…Detection of somatic mutations in cancer, genes responsible for hereditary diseases, polymorphisms and other applications are described in detail at Kakavas et al [ 8 – 10 ]. Recent researches in the last 4 years shows dispersion of SSCP method in the above applications: on cancer prognosis [ 11 , 12 ], on asthma disease [ 13 ], on blood groups [ 14 ], on gilbert’s syndrome [ 15 ], on diabetes disease [ 16 ], on respiratory distress syndrome [ 17 ], on infertile men with varicocele [ 18 ], on gastric mucosa [ 19 ], on traditional Chinese medicine [ 20 ], detection of bacterial DNA [ 21 ], identification of trichomonas vaginalis [ 22 ].…”

Section: Medical Diagnosis Applicationsmentioning

confidence: 99%

Sensitivity and applications of the PCR Single-Strand Conformation Polymorphism method

Kakavas

2021

Mol Biol Rep

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“…Briefly, the SSCP method is based on the observation that, under non-denaturing conditions, single stranded DNA (ssDNA) fragments fall into unique conformations (Hayashi, 1992), determined by their primary sequence, in structures stabilized by intramolecular interactions (Orita et al, 1989;Kakavas et al, 2006). As a consequence, even a single base alteration can result in a conformational change which can be detected by the altered mobility of the singlestranded DNA molecule in SSCP.…”

Section: Introductionmentioning

confidence: 99%

Identification of the commonest cystic fibrosis transmembrane regulator gene ΔF508 mutation: evaluation of PCR–single‐strand conformational polymorphism and polyacrylamide gel electrophoresis

Kakavas

Noulas

Kanakis

et al. 2006

Biomedical Chromatography

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In the present study we investigated whether single-strand conformational polymorphism (SSCP) and polyacrylamide gel electrophoresis (PAGE) could be used for the identification of the CFTR DeltaF508 gene mutation, which is commonest in the Greek population. Using DNA from patients carrying this mutation, the appropriate 98 bp region of the CFTR gene was amplified by PCR and the reaction products were analysed by non-radioactive SSCP-electrophoresis using silver staining for band visualization and non-denaturating PAGE to confirm the results. SSCP electrophoretic analysis has been optimized for several parameters in order to achieve the best resolution. Single-strand DNA fragments gave a reproducible pattern of bands, characteristic for the particular mutation. Comparison of the obtain patterns with control samples allowed the detection of the DeltaF508 mutation in the patients studied by SSCP assay and these results were confirmed by the independent method of PAGE. Although SSCP and PAGE can be used for detection of this mutation, PAGE resulted in more distinct patterns than SSCP. It is, therefore, proposed that PAGE can be reliably used for the detection and identification of such a mutation in patients provided that suitable controls are available. The applicability of PAGE to identification of the mutation in carriers, particularly useful for population screening, is also discussed.

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Identification of the four most common β‐globin gene mutations in Greek β‐thalassemic patients and carriers by PCR‐SSCP: advantages and limitations of the method

Cited by 5 publications

References 20 publications

PCR–SSCP: A Method for the Molecular Analysis of Genetic Diseases

PCR–SSCP: A Method for the Molecular Analysis of Genetic Diseases

Sensitivity and applications of the PCR Single-Strand Conformation Polymorphism method

Identification of the commonest cystic fibrosis transmembrane regulator gene ΔF508 mutation: evaluation of PCR–single‐strand conformational polymorphism and polyacrylamide gel electrophoresis

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