Using immunofluorescence (antigen) mapping in the diagnosis and classification of epidermolysis bullosa: a first report from Iran

Int J Dermatol. 2015 Oct;54(10):e416-23. doi: 10.1111/ijd.12804. Epub 2015 Jul 28.

Abstract

Background: Immunofluorescence antigen mapping (IFM), is a newly introduced technique for diagnosis and classification of epidermolysis bullosa (EB) disease. The precise level of skin cleavage can be determined using monoclonal antibodies to EB-specific basement membrane zone protein.

Objective: To apply IFM technique in diagnosis and classification of EB and to identify utility and limitation of this method in our clinical setting.

Methods: IFM was done according to a described protocol by Pohla-Gubo et al. Monoclonal antibodies used for antigen mapping were against cytokeratin 5, cytokeratin 14, α6 integrin, β4 integrin, laminin 332, Collagen IV, and Collagen VII.

Results: IFM was done for 95 referred patients, compromising 49 females and 46 males, aged 5 days to 45 years (mean = 9.5 years). Ninety cases were diagnosed with EB and classified as follows: EB simplex: (n = 13), junctional EB (n = 14), dystrophic EB (n = 62), and Kindler syndrome (n = 1). Diagnosis was not made in five cases as their specimens contained no blister. Confirmatory genetic analysis was done for five junctional cases from two families with clinical features of laryngo-onycho-cutaneous syndrome. Genetic molecular studies showed nonsense mutations in the last codon of exon 39 of the laminin α3a (LAMA3) gene (p.Gln57X) and a donor splice site mutation in LAMA3 (IVS57+5G>A) in the first and second family, respectively.

Conclusion: IFM technique is relatively simple to perform, and interpretation of the results is not sophisticated. The proportion of inconclusive results will be decreased if the specimens contain freshly induced blister.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Antibodies, Monoclonal
  • Blister / diagnosis
  • Child
  • Child, Preschool
  • Codon, Nonsense
  • Collagen Type IV / immunology
  • Collagen Type VII / immunology
  • Epidermolysis Bullosa / classification*
  • Epidermolysis Bullosa / diagnosis*
  • Epidermolysis Bullosa Dystrophica / diagnosis
  • Epidermolysis Bullosa Simplex / diagnosis
  • Epidermolysis Bullosa, Junctional / diagnosis
  • Epidermolysis Bullosa, Junctional / genetics
  • Female
  • Fluorescent Antibody Technique / methods*
  • Humans
  • Infant
  • Infant, Newborn
  • Integrin alpha6 / immunology
  • Integrin beta4 / immunology
  • Iran
  • Keratin-14 / immunology
  • Keratin-5 / immunology
  • Laminin / genetics
  • Laminin / immunology
  • Male
  • Middle Aged
  • Periodontal Diseases / diagnosis
  • Photosensitivity Disorders / diagnosis
  • Young Adult

Substances

  • Antibodies, Monoclonal
  • Codon, Nonsense
  • Collagen Type IV
  • Collagen Type VII
  • Integrin alpha6
  • Integrin beta4
  • Keratin-14
  • Keratin-5
  • Laminin
  • laminin alpha 3

Supplementary concepts

  • Poikiloderma of Kindler