Two novel nonradioactive polymerase chain reaction-based assays of dried blood spots, genomic DNA, or whole cells for fast, reliable detection of Z and S mutations in the alpha 1-antitrypsin gene

B S Andresen, I Knudsen, P K Jensen, K Rasmussen, N Gregersen

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

Resumé

Two new nonradioactive polymerase chain reaction (PCR)-based assays for the Z and S mutations in the alpha 1-antitrypsin gene are presented. The assays take advantage of PCR-mediated mutagenesis, creating new diagnostic restriction enzyme sites for unambiguous discrimination between test samples from individuals who are normal, heterozygous, or homozygous for the mutations. We show that the two assays can be performed with purified genomic DNA as well as with boiled blood spots. The new assays were validated by parallel testing with a technique in which PCR is combined with allele-specific oligonucleotide (ASO) probes. In all cases tested the results obtained by the different techniques were in accordance. The new assays can be used for prenatal diagnostics and can be performed directly with boiled tissue samples. Because the new assays are easy to perform and reliable, we conclude that they are well suited for routine diagnosis.

OriginalsprogEngelsk
TidsskriftClinical Chemistry
Vol/bind38
Udgave nummer10
Sider (fra-til)2100-7
Antal sider8
ISSN0009-9147
StatusUdgivet - 1992

Fingeraftryk

alpha 1-Antitrypsin
Polymerase chain reaction
Assays
Blood
Genes
Polymerase Chain Reaction
Mutation
DNA
Oligonucleotide Probes
Alleles
Mutagenesis
Enzymes
Tissue
Testing

Citer dette

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abstract = "Two new nonradioactive polymerase chain reaction (PCR)-based assays for the Z and S mutations in the alpha 1-antitrypsin gene are presented. The assays take advantage of PCR-mediated mutagenesis, creating new diagnostic restriction enzyme sites for unambiguous discrimination between test samples from individuals who are normal, heterozygous, or homozygous for the mutations. We show that the two assays can be performed with purified genomic DNA as well as with boiled blood spots. The new assays were validated by parallel testing with a technique in which PCR is combined with allele-specific oligonucleotide (ASO) probes. In all cases tested the results obtained by the different techniques were in accordance. The new assays can be used for prenatal diagnostics and can be performed directly with boiled tissue samples. Because the new assays are easy to perform and reliable, we conclude that they are well suited for routine diagnosis.",
keywords = "Base Sequence, Cells, Cultured, Chorionic Villi, DNA, Deoxyribonucleases, Type II Site-Specific, Female, Fetus, Heterozygote, Homozygote, Humans, Molecular Sequence Data, Mutation, Placenta, Polymerase Chain Reaction, Pregnancy, Prenatal Diagnosis, alpha 1-Antitrypsin, alpha 1-Antitrypsin Deficiency",
author = "Andresen, {B S} and I Knudsen and Jensen, {P K} and K Rasmussen and N Gregersen",
year = "1992",
language = "English",
volume = "38",
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journal = "Clinical Chemistry",
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Two novel nonradioactive polymerase chain reaction-based assays of dried blood spots, genomic DNA, or whole cells for fast, reliable detection of Z and S mutations in the alpha 1-antitrypsin gene. / Andresen, B S; Knudsen, I; Jensen, P K; Rasmussen, K; Gregersen, N.

I: Clinical Chemistry, Bind 38, Nr. 10, 1992, s. 2100-7.

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningpeer review

TY - JOUR

T1 - Two novel nonradioactive polymerase chain reaction-based assays of dried blood spots, genomic DNA, or whole cells for fast, reliable detection of Z and S mutations in the alpha 1-antitrypsin gene

AU - Andresen, B S

AU - Knudsen, I

AU - Jensen, P K

AU - Rasmussen, K

AU - Gregersen, N

PY - 1992

Y1 - 1992

N2 - Two new nonradioactive polymerase chain reaction (PCR)-based assays for the Z and S mutations in the alpha 1-antitrypsin gene are presented. The assays take advantage of PCR-mediated mutagenesis, creating new diagnostic restriction enzyme sites for unambiguous discrimination between test samples from individuals who are normal, heterozygous, or homozygous for the mutations. We show that the two assays can be performed with purified genomic DNA as well as with boiled blood spots. The new assays were validated by parallel testing with a technique in which PCR is combined with allele-specific oligonucleotide (ASO) probes. In all cases tested the results obtained by the different techniques were in accordance. The new assays can be used for prenatal diagnostics and can be performed directly with boiled tissue samples. Because the new assays are easy to perform and reliable, we conclude that they are well suited for routine diagnosis.

AB - Two new nonradioactive polymerase chain reaction (PCR)-based assays for the Z and S mutations in the alpha 1-antitrypsin gene are presented. The assays take advantage of PCR-mediated mutagenesis, creating new diagnostic restriction enzyme sites for unambiguous discrimination between test samples from individuals who are normal, heterozygous, or homozygous for the mutations. We show that the two assays can be performed with purified genomic DNA as well as with boiled blood spots. The new assays were validated by parallel testing with a technique in which PCR is combined with allele-specific oligonucleotide (ASO) probes. In all cases tested the results obtained by the different techniques were in accordance. The new assays can be used for prenatal diagnostics and can be performed directly with boiled tissue samples. Because the new assays are easy to perform and reliable, we conclude that they are well suited for routine diagnosis.

KW - Base Sequence

KW - Cells, Cultured

KW - Chorionic Villi

KW - DNA

KW - Deoxyribonucleases, Type II Site-Specific

KW - Female

KW - Fetus

KW - Heterozygote

KW - Homozygote

KW - Humans

KW - Molecular Sequence Data

KW - Mutation

KW - Placenta

KW - Polymerase Chain Reaction

KW - Pregnancy

KW - Prenatal Diagnosis

KW - alpha 1-Antitrypsin

KW - alpha 1-Antitrypsin Deficiency

M3 - Journal article

C2 - 1394999

VL - 38

SP - 2100

EP - 2107

JO - Clinical Chemistry

JF - Clinical Chemistry

SN - 0009-9147

IS - 10

ER -