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Mistyping of the human angiotensin-converting enzyme gene polymorphism: frequency, causes and possible methods to avoid errors in typing
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Description
<jats:title>ABSTRACT</jats:title> <jats:p>A polymorphism of the gene encoding the human angiotensin I-converting enzyme (ACE), which is defined by an insertion/deletion polymorphism in intron 16, has been identified as a candidate genetic locus in the development of cardiovascular and renal disease.</jats:p> <jats:p>We have demonstrated that the accuracy of ACE genotyping is critically dependent on the strategy of the PCR used in typing. Of 1238 individuals genotyped by a standard method, 335 were typed as DD, 645 as DI and 258 as II. However, when DD individuals were retyped using modified methods (including either 5% dimethyl sulphoxide, or a 'hot start') 35 of the original 335 samples (10·5%) were retyped as DI.</jats:p> <jats:p>In approximately half of these mistyped samples, PCR amplification was assessed as inefficient by the absence of a third faint heteroduplex band in a control ID sample: when the assay was repeated without any modifications, the mistyped samples were correctly genotyped. In the remainder, mistyping persisted. In these cases, the use of a third 'nested' PCR primer specific for the I allele was required for successful genotyping, providing a more reliable strategy without the need for further modification to the PCR technique. Our results suggest that the triple primer approach is the method of choice for accurate ACE genotyping.</jats:p>
Journal
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- Journal of Molecular Endocrinology
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Journal of Molecular Endocrinology 17 (1), 27-30, 1996-08
Bioscientifica
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Details 詳細情報について
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- CRID
- 1363107371176313984
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- ISSN
- 14796813
- 09525041
- http://id.crossref.org/issn/00220795
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- Data Source
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- Crossref