Evaluation of 'at risk' alpha 1-antitrypsin genotype SZ with synthetic oligonucleotide gene probes

T. Nukiwa, M. Brantly, R. Garver, L. Paul, M. Courtney, J. P. LeCocq, Ronald Crystal

Research output: Contribution to journalArticle

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Abstract

Alpha 1-antitrypsin (α1AT), a 52,000-mol-wt serum glycoprotein produced by hepatocytes and mononuclear phagocytes, functions as the major inhibitor of neutrophil elastase. The α1AT haplotype S is associated with childhood liver disease and/or adult emphysema when inherited with the Z haplotype to give the phenotype SZ. To accurately identify the SZ phenotype at the level of genomic DNA, four 32P-labeled 19-mer synthetic oligonucleotide probes were prepared; two to identify the M and S difference in exon III, and two to identify the M and Z difference in exon V. These probes were hybridized with various cloned DNAs and genomic DNAs cut with the restriction endonucleases BglI and EcoRI; the genomic DNAs represented all six possible phenotype combinations of the M, S, and Z haplotypes (MM, MS, MZ, SS, ZZ, and SZ). Using the four probes to evaluate 42 samples of genomic DNA, the 'at risk' SZ and ZZ phenotypes were correctly identified in all cases, as were the 'not at risk' phenotypes SS, MS, MM, and MZ, demonstrating that both exon III and exon V directed probes are necessary to properly identify all of the major 'at risk' α1AT genes. However, when used to evaluate a very rare family carrying a null allele, these four oligonucleotide probes misindentified the 'at risk' null-null and S null phenotypes as 'not at risk' MM and SM combinations. These observations indicate that oligonucleotide gene probes yielded reliable and accurate assessment of 'at risk' α1AT genotypes in almost all situations, but in the context of prenatal diagnosis and genetic counseling this approach must be used with caution and in combination with family studies so as not to misidentify rare genotypes that may be associated with a risk for disease.

Original languageEnglish
Pages (from-to)528-537
Number of pages10
JournalJournal of Clinical Investigation
Volume77
Issue number2
Publication statusPublished - 15 Oct 1986
Externally publishedYes

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Synthetic Genes
alpha 1-Antitrypsin
Oligonucleotide Probes
Genotype
Phenotype
Exons
DNA
Haplotypes
Secretory Proteinase Inhibitory Proteins
Deoxyribonuclease EcoRI
DNA Restriction Enzymes
Genetic Counseling
Phagocytes
Prenatal Diagnosis
Genes
Liver Diseases
Hepatocytes
Glycoproteins
Alleles
Serum

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Nukiwa, T., Brantly, M., Garver, R., Paul, L., Courtney, M., LeCocq, J. P., & Crystal, R. (1986). Evaluation of 'at risk' alpha 1-antitrypsin genotype SZ with synthetic oligonucleotide gene probes. Journal of Clinical Investigation, 77(2), 528-537.

Evaluation of 'at risk' alpha 1-antitrypsin genotype SZ with synthetic oligonucleotide gene probes. / Nukiwa, T.; Brantly, M.; Garver, R.; Paul, L.; Courtney, M.; LeCocq, J. P.; Crystal, Ronald.

In: Journal of Clinical Investigation, Vol. 77, No. 2, 15.10.1986, p. 528-537.

Research output: Contribution to journalArticle

Nukiwa, T, Brantly, M, Garver, R, Paul, L, Courtney, M, LeCocq, JP & Crystal, R 1986, 'Evaluation of 'at risk' alpha 1-antitrypsin genotype SZ with synthetic oligonucleotide gene probes', Journal of Clinical Investigation, vol. 77, no. 2, pp. 528-537.
Nukiwa T, Brantly M, Garver R, Paul L, Courtney M, LeCocq JP et al. Evaluation of 'at risk' alpha 1-antitrypsin genotype SZ with synthetic oligonucleotide gene probes. Journal of Clinical Investigation. 1986 Oct 15;77(2):528-537.
Nukiwa, T. ; Brantly, M. ; Garver, R. ; Paul, L. ; Courtney, M. ; LeCocq, J. P. ; Crystal, Ronald. / Evaluation of 'at risk' alpha 1-antitrypsin genotype SZ with synthetic oligonucleotide gene probes. In: Journal of Clinical Investigation. 1986 ; Vol. 77, No. 2. pp. 528-537.
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