Novel Diagnostic Tool for p47 ^phox -Deficient Chronic Granulomatous Disease Patient and Carrier Detection.

ABSTRACT: Chronic granulomatous disease (CGD) is a primary immunodeficiency caused by mutations of the phagocytic nicotinamide adenine dinucleotide phosphate (NADPH) oxidase. Autosomal recessive p47 ^phox -deficient CGD (p47 ^phox CGD) is the second most frequent form of the disease in western countries, and more than 94% of patients have a disease-causing dinucleotide deletion (ΔGT) in the neutrophil cytosolic factor 1 (NCF1) gene. The ΔGT mutation is most likely transferred onto the NCF1 from one of its two pseudogenes co-localized on the same chromosome. The presence of NCF1 pseudogenes in healthy individuals makes the genetic diagnostics of ΔGT p47 ^phox CGD challenging, as it requires the distinction between ΔGT in NCF1 and in the two pseudogenes. We have developed a diagnostic tool for the identification of p47 ^phox CGD based on PCR co-amplification of NCF1 and its pseudogenes, followed by band intensity quantification of restriction fragment length polymorphism products. The single-day, reliable p47 ^phox CGD diagnostics allow for robust discrimination of homozygous ΔGT p47^phox CGD patients from heterozygous carriers and healthy individuals, as well as for monitoring gene therapy efficacy.

SUBMITTER: Wrona D 

PROVIDER: S-EPMC6395829 | biostudies-literature | 2019 Jun

REPOSITORIES: biostudies-literature