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Title: Rapid deoxyribonucleic acid analysis by allele-specific polymerase chain reaction for detection of mutations in the steroid 21-hydroxylase gene

Abstract

Rapid DNA analysis based on allele-specific polymerase chain reaction (PCR) using mutation site-specific primers was developed to detect mutations in the CYP21 gene known to cause steroid 21-hydroxylase deficiency. In contrast to the previous method, in which PCR of genomic DNA was followed by dot blot analysis with radio active probes and multiple rounds of stripping and reprobing for each of the 8 most common mutation sites, the results using this new method were immediately visualized after the PCR run by ethidium bromide-stained agarose gel electrophoresis. Using allele-specific PCR, mutation(s) were identified on 148 affected chromosomes out of 160 tested. Although mutation(s) were identified on only one chromosome of 11 of these patients, their parents showed a consistent pattern on DNA analysis. The only exception was that in one family, in which the parents each had a detectable mutation, a mutation was detected on only one allele of the patient. Most likely there is a mutation in the patient`s other allele that could have arisen de novo or was inherited from the parent and was not evident in the transmitting parent`s phenotype. When compared with the dot blot procedure, allele-specific PCR is more rapid, less labor-intensive, and avoids the usemore » of radioactivity. 26 refs., 3 figs., 2 tabs.« less

Authors:
; ;  [1]
  1. New York Hospital-Cornell Medical Center, NY (United States); and others
Publication Date:
OSTI Identifier:
283132
Resource Type:
Journal Article
Journal Name:
Journal of Clinical Endocrinology and Metabolism
Additional Journal Information:
Journal Volume: 80; Journal Issue: 5; Other Information: PBD: May 1995
Country of Publication:
United States
Language:
English
Subject:
55 BIOLOGY AND MEDICINE, BASIC STUDIES; GENES; GENE MUTATIONS; STRUCTURE-ACTIVITY RELATIONSHIPS; POLYMERASE CHAIN REACTION; EVALUATION; DNA; QUALITATIVE CHEMICAL ANALYSIS; DETECTION; STEROIDS; HYDROXYLASES; ELECTROPHORESIS; PROBES; EXONS

Citation Formats

Wilson, R C, Wei, J Q, and Cheng, K C. Rapid deoxyribonucleic acid analysis by allele-specific polymerase chain reaction for detection of mutations in the steroid 21-hydroxylase gene. United States: N. p., 1995. Web. doi:10.1210/jc.80.5.1635.
Wilson, R C, Wei, J Q, & Cheng, K C. Rapid deoxyribonucleic acid analysis by allele-specific polymerase chain reaction for detection of mutations in the steroid 21-hydroxylase gene. United States. https://doi.org/10.1210/jc.80.5.1635
Wilson, R C, Wei, J Q, and Cheng, K C. 1995. "Rapid deoxyribonucleic acid analysis by allele-specific polymerase chain reaction for detection of mutations in the steroid 21-hydroxylase gene". United States. https://doi.org/10.1210/jc.80.5.1635.
@article{osti_283132,
title = {Rapid deoxyribonucleic acid analysis by allele-specific polymerase chain reaction for detection of mutations in the steroid 21-hydroxylase gene},
author = {Wilson, R C and Wei, J Q and Cheng, K C},
abstractNote = {Rapid DNA analysis based on allele-specific polymerase chain reaction (PCR) using mutation site-specific primers was developed to detect mutations in the CYP21 gene known to cause steroid 21-hydroxylase deficiency. In contrast to the previous method, in which PCR of genomic DNA was followed by dot blot analysis with radio active probes and multiple rounds of stripping and reprobing for each of the 8 most common mutation sites, the results using this new method were immediately visualized after the PCR run by ethidium bromide-stained agarose gel electrophoresis. Using allele-specific PCR, mutation(s) were identified on 148 affected chromosomes out of 160 tested. Although mutation(s) were identified on only one chromosome of 11 of these patients, their parents showed a consistent pattern on DNA analysis. The only exception was that in one family, in which the parents each had a detectable mutation, a mutation was detected on only one allele of the patient. Most likely there is a mutation in the patient`s other allele that could have arisen de novo or was inherited from the parent and was not evident in the transmitting parent`s phenotype. When compared with the dot blot procedure, allele-specific PCR is more rapid, less labor-intensive, and avoids the use of radioactivity. 26 refs., 3 figs., 2 tabs.},
doi = {10.1210/jc.80.5.1635},
url = {https://www.osti.gov/biblio/283132}, journal = {Journal of Clinical Endocrinology and Metabolism},
number = 5,
volume = 80,
place = {United States},
year = {1995},
month = {5}
}