Detection of hemizygosity in Hermansky - Pudlak syndrome by quantitative real-time PCR

A. E. Griffin, B. R. Cobb, P. D. Anderson, D. A. Claassen, A. Helip-Wooley, M. Huizing, William A. Gahl

Research output: Contribution to journalArticle

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Abstract

Hermansky - Pudlak syndrome (HPS) is an autosomal recessive disorder characterized by oculocutaneous albinism, a bleeding diathesis and, in some patients, pulmonary fibrosis or granulomatous colitis. HPS is associated with biosynthesis defects of melanosomes, platelet-dense bodies, and lysosomes. There are seven genetic HPS subtypes; HPS-1 is the most common. We used a real-time quantitative PCR (qPCR) approach to investigate six HPS-1 patients, previously assigned as having homozygous mutations in the HPS1 gene. HPS1 gene copy numbers, calculated by use of a comparative Ct method, revealed that one patient was in fact hemizygous for her c.1189delC (S396delC) HPS1 mutation. The causative deletion/insertion was 13,966 bp in size, with defined breakpoints, and involved an adjacent gene (C10orf33). A mechanism of formation is proposed for the deletion/insertion, and both multiplex and qPCR indicated that the deletion/insertion was present in the patient, her brother, and her father. qPCR amplification is valuable for detecting deletions too small to be identified by fluorescence in situ hybridization. This demonstration of hemizygosity, performed using genomic DNA, can eliminate concerns about non-paternity and can verify the diagnosis of an autosomal recessive disorder when a DNA alteration appears to be homozygous by standard PCR and sequencing methods, and its pathogenicity is in doubt.

Original languageEnglish (US)
Pages (from-to)23-30
Number of pages8
JournalClinical Genetics
Volume68
Issue number1
DOIs
StatePublished - Jul 2005

Fingerprint

Hermanski-Pudlak Syndrome
Real-Time Polymerase Chain Reaction
Oculocutaneous Albinism
Melanosomes
Polymerase Chain Reaction
Mutation
Gene Dosage
Pulmonary Fibrosis
Multiplex Polymerase Chain Reaction
DNA
Disease Susceptibility
Lysosomes
Fluorescence In Situ Hybridization
Fathers
Crohn Disease
Genes
Virulence
Siblings
Blood Platelets
Hemorrhage

Keywords

  • Breakpoint
  • C10orf33
  • Genomic insertion/deletion
  • Hemizygosity
  • Hermansky-Pudlak syndrome
  • Real-time quantitative PCR

ASJC Scopus subject areas

  • Genetics(clinical)
  • Genetics

Cite this

Griffin, A. E., Cobb, B. R., Anderson, P. D., Claassen, D. A., Helip-Wooley, A., Huizing, M., & Gahl, W. A. (2005). Detection of hemizygosity in Hermansky - Pudlak syndrome by quantitative real-time PCR. Clinical Genetics, 68(1), 23-30. https://doi.org/10.1111/j.1399-0004.2005.00461.x

Detection of hemizygosity in Hermansky - Pudlak syndrome by quantitative real-time PCR. / Griffin, A. E.; Cobb, B. R.; Anderson, P. D.; Claassen, D. A.; Helip-Wooley, A.; Huizing, M.; Gahl, William A.

In: Clinical Genetics, Vol. 68, No. 1, 07.2005, p. 23-30.

Research output: Contribution to journalArticle

Griffin, AE, Cobb, BR, Anderson, PD, Claassen, DA, Helip-Wooley, A, Huizing, M & Gahl, WA 2005, 'Detection of hemizygosity in Hermansky - Pudlak syndrome by quantitative real-time PCR', Clinical Genetics, vol. 68, no. 1, pp. 23-30. https://doi.org/10.1111/j.1399-0004.2005.00461.x
Griffin AE, Cobb BR, Anderson PD, Claassen DA, Helip-Wooley A, Huizing M et al. Detection of hemizygosity in Hermansky - Pudlak syndrome by quantitative real-time PCR. Clinical Genetics. 2005 Jul;68(1):23-30. https://doi.org/10.1111/j.1399-0004.2005.00461.x
Griffin, A. E. ; Cobb, B. R. ; Anderson, P. D. ; Claassen, D. A. ; Helip-Wooley, A. ; Huizing, M. ; Gahl, William A. / Detection of hemizygosity in Hermansky - Pudlak syndrome by quantitative real-time PCR. In: Clinical Genetics. 2005 ; Vol. 68, No. 1. pp. 23-30.
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