Rom J Morphol Embryol 2011, 52(2):919–922 CASE REPORTS Dystrophic epidermolysis bullosa: two case reports SORINA DĂNESCU 1) , SIMONA ŞENILĂ 1) , LUMINIŢA LELUŢIU 2) , S. NEDEVSCHI 3) , RODICA COSGAREA 1) 1) “Iuliu Haţieganu” University of Medicine and Pharmacy, Cluj-Napoca 2) “Prof. dr. Ion Chiricuţă” Oncology Institute, Cluj-Napoca 3) Technical University of Cluj-Napoca Abstract We identified the mutations in two patients with different phenotypes of dystrophic epidermolysis bullosa (DEB). We performed molecular diagnosis to a patient aged 45 years who showed the typical severe generalized autosomal recessive DEB signs when admitted to the hospital. The other patient is a 4-month-old boy who showed a moderate clinical aspect of DEB, dominated by nail dystrophy. The molecular diagnosis disclosed in the first patient the presence of a heterozygous mutation consisting of a nucleotide substitution that lead to a splice site mutation, namely 425-2 A>G, associated to a premature termination codon, in exon 5, namely c.553 C>T, p.R185X and in the second patient a heterozygous substitution at nucleotide position 6100 that converts a glycine amino acid to arginine (6100G>A). The mutation is designated G2034R. We conclude that molecular diagnosis is the conclusive EBD investigation, maps the phenotype of a patient with his genotype and thus allows a better understanding of the disease mechanism and the development of gene therapy. Molecular diagnosis also enables genetic counseling and prenatal diagnosis. Keywords: dystrophic epidermolysis bullosa, molecular diagnosis, type VII collagen gene mutation. Introduction Epidermolysis bullosa (EB) is a heterogeneous group of heritable skin disorders, characterized by blistering of the skin and of the mucous membranes, following minor trauma [1]. Dystrophic EB is caused by mutations in COL7A1, the gene coding for collagen VII, the major component of the anchoring fibrils. The third International Consensus Meeting on Diagnosis and Classification of EB classified dystrophic EB into two types: DDEB (dominant dystrophic epidermolysis bullosa) and RDEB (recessive dystrophic epidermolysis bullosa) with their subtypes. The sub- types of DDEB are generalized DDEB, acral DDEB, pretibial DDEB, DDEB pruriginosa, DDEB nails only and DDEB bullous dermolysis of the newborn. RDEB is subdivided into severe, generalized RDEB (the new name for Hallopeau–Siemens); generalized RDEB, other (the new name for non-Hallopeau– Siemens); inversa RDEB; pretibial RDEB, pruriginosa RDEB; centripetalis RDEB; bullous dermolysis RDEB of the newborn [2]. Materials and Methods After informed consent, genomic DNA was extracted from a peripheral blood sample from the affected patients. Genomic DNA was extracted from EDTA blood samples using a DNA isolation kit according to the manufacturer’s protocol. Polymerase chain reaction (PCR) amplification was carried out. For sequencing the same primers as for PCR were used and the products were sequenced directly on an 8-capilary genetic analyzer. Results Case No. 1 The first case was a female patient aged 45 years who showed the typical signs of RDEB – severe generalized when admitted to the hospital. The clinical features included ulceration, severe scars that caused mutilations of hands and feet, fibrosis of the oral cavity, weight loss, edentia, esophageal dysmotility and strictures, anemia, constipation and squamous cell carcinoma (Figures 1–3). A biopsy from the lesion suspected as squamous cell carcinoma was prelevated and the diagnostic was confirmed by the histopathological examination, which shows a proliferation of lobules of glassy, eosinophilic keratino- cytes and keratin pearls (Figure 4). No other family member showed any cutaneous or extracutaneous lesions and her parents were not consanguine. The treatment was based on skin care. We used special dressings, which comprises a silicone mesh, providing a good rate of epithelization. Bacterial infection was treated with systemic antibacterial agents, according to the antibiogram. The food was liquidized and the anemia was treated with iron supplements. The patient refused surgical treatment for squamous cell carcinoma. The molecular diagnosis disclosed the presence of a heterozygous mutation consisting of a nucleotide R J M E Romanian Journal of Morphology & Embryology http://www.rjme.ro/