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Multiplex PCR analysis and genotype-phenotype correlations of frequent APC mutations.
Hum Mutat. 1995; 5(2):144-52.HM

Abstract

Germline mutations of the adenomatous polyposis coli (APC) gene tend to cluster in discrete regions. Some of these mutations occur frequently in familial adenomatous polyposis coli (FAP) patients, and strategies for genetic diagnosis of the disease should include simple methods for their detection. We studied a total of 48 FAP-affected or "at-risk" members from 31 unrelated FAP pedigrees. Unrelated patients were analyzed using heteroduplex analysis on agarose minigels (HAAM) and multiplex allele-specific PCR. This novel strategy readily and reliably detected the three frequently occurring APC deletions at codons 1061, 1068, and 1309, allowing identification of mutant alleles in nine unrelated patients. A targeted mutational analysis, based on HAAM and amplification refractory mutation system (ARMS), allowed the rapid identification of 11 additional subjects with germline deletions, among relatives of the patients in whom mutations had been detected by multiplex PCR and HAAM. The use of two independent PCR-based tests, employing distinct sets of primers, reduces the possibility that artifacts occurring during DNA amplification may interfere with the diagnostic evaluation. The analysis of genotype-phenotype correlations provided evidence for heterogeneity with regard to the extent of colonic and extracolonic manifestations of the disease in subjects bearing identical mutations. However, the consistent association of the deletion at codon 1309 with more severe colonic disease than that observed in patients with mutations at codons 1061 and 1068, supports a correlation between mutation site and penetrance of FAP.

Authors+Show Affiliations

Istituto di Patologia Umana e Medicina Sociale, Università Gabriele D'Annunzio, Chieti, Italy.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

7749413

Citation

Cama, A, et al. "Multiplex PCR Analysis and Genotype-phenotype Correlations of Frequent APC Mutations." Human Mutation, vol. 5, no. 2, 1995, pp. 144-52.
Cama A, Palmirotta R, Curia MC, et al. Multiplex PCR analysis and genotype-phenotype correlations of frequent APC mutations. Hum Mutat. 1995;5(2):144-52.
Cama, A., Palmirotta, R., Curia, M. C., Esposito, D. L., Ranieri, A., Ficari, F., Valanzano, R., Battista, P., Modesti, A., & Tonelli, F. (1995). Multiplex PCR analysis and genotype-phenotype correlations of frequent APC mutations. Human Mutation, 5(2), 144-52.
Cama A, et al. Multiplex PCR Analysis and Genotype-phenotype Correlations of Frequent APC Mutations. Hum Mutat. 1995;5(2):144-52. PubMed PMID: 7749413.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Multiplex PCR analysis and genotype-phenotype correlations of frequent APC mutations. A1 - Cama,A, AU - Palmirotta,R, AU - Curia,M C, AU - Esposito,D L, AU - Ranieri,A, AU - Ficari,F, AU - Valanzano,R, AU - Battista,P, AU - Modesti,A, AU - Tonelli,F, PY - 1995/1/1/pubmed PY - 1995/1/1/medline PY - 1995/1/1/entrez SP - 144 EP - 52 JF - Human mutation JO - Hum Mutat VL - 5 IS - 2 N2 - Germline mutations of the adenomatous polyposis coli (APC) gene tend to cluster in discrete regions. Some of these mutations occur frequently in familial adenomatous polyposis coli (FAP) patients, and strategies for genetic diagnosis of the disease should include simple methods for their detection. We studied a total of 48 FAP-affected or "at-risk" members from 31 unrelated FAP pedigrees. Unrelated patients were analyzed using heteroduplex analysis on agarose minigels (HAAM) and multiplex allele-specific PCR. This novel strategy readily and reliably detected the three frequently occurring APC deletions at codons 1061, 1068, and 1309, allowing identification of mutant alleles in nine unrelated patients. A targeted mutational analysis, based on HAAM and amplification refractory mutation system (ARMS), allowed the rapid identification of 11 additional subjects with germline deletions, among relatives of the patients in whom mutations had been detected by multiplex PCR and HAAM. The use of two independent PCR-based tests, employing distinct sets of primers, reduces the possibility that artifacts occurring during DNA amplification may interfere with the diagnostic evaluation. The analysis of genotype-phenotype correlations provided evidence for heterogeneity with regard to the extent of colonic and extracolonic manifestations of the disease in subjects bearing identical mutations. However, the consistent association of the deletion at codon 1309 with more severe colonic disease than that observed in patients with mutations at codons 1061 and 1068, supports a correlation between mutation site and penetrance of FAP. SN - 1059-7794 UR - https://www.unboundmedicine.com/medline/citation/7749413/Multiplex_PCR_analysis_and_genotype_phenotype_correlations_of_frequent_APC_mutations_ L2 - https://doi.org/10.1002/humu.1380050208 DB - PRIME DP - Unbound Medicine ER -