Copy number analysis of complement C4A, C4B and C4A silencing mutation by real-time quantitative polymerase chain reaction
| Autor: | Katja T. Eronen, Riitta Paakkanen, Hanna Vauhkonen, Marja-Liisa Lokki, Mikko Seppänen, Asko Järvinen |
|---|---|
| Přispěvatelé: | Haartman Institute (-2014), Transplantation Laboratory, Kardiologian yksikkö, Department of Medicine, Infektiosairauksien yksikkö, Clinicum |
| Jazyk: | angličtina |
| Rok vydání: | 2012 |
| Předmět: |
Low protein
Complement System Gene Dosage COMPONENTS C4A Polymerase Chain Reaction DISEASE 0302 clinical medicine Gene Frequency Gene Duplication Genetics of the Immune System Copy-number variation SYSTEMIC-LUPUS-ERYTHEMATOSUS RCCX MODULES Genetics 0303 health sciences Multidisciplinary Complement C4a Null allele Innate Immunity Clinical Laboratory Sciences 3. Good health DEFICIENCY Real-time polymerase chain reaction Infectious Diseases Phenotype 030220 oncology & carcinogenesis POPULATIONS Medicine Research Article DNA Copy Number Variations Science Immunology education Copy number analysis Biology Real-Time Polymerase Chain Reaction Gene dosage Sensitivity and Specificity Autoimmune Diseases Immunophenotyping Molecular Genetics 03 medical and health sciences Immune Deficiency Genetic Mutation Diagnostic Medicine Complement C4b Humans Genetic Testing Gene Silencing Allele frequency NULL ALLELES Genetic Association Studies Alleles 030304 developmental biology DNA Primers Clinical Genetics Models Genetic C4A Personalized Medicine Immunity Reproducibility of Results Human Genetics Molecular biology GENE POLYMORPHISM Gene Expression Regulation Immune System 3121 General medicine internal medicine and other clinical medicine Genetics of Disease Mutation RISK-FACTORS Clinical Immunology 3111 Biomedicine |
| Zdroj: | PLoS ONE, Vol 7, Iss 6, p e38813 (2012) PLoS ONE |
| ISSN: | 1932-6203 |
| Popis: | Low protein levels and copy number variation (CNV) of the fourth component of human complement (C4A and C4B) have been associated with various diseases. High-throughput methods for analysing C4 CNV are available, but they commonly do not detect the most common C4A mutation, a silencing CT insertion (CTins) leading to low protein levels. We developed a SYBR® Green labelled real-time quantitative polymerase chain reaction (qPCR) with a novel concentration range approach to address C4 CNV and deficiencies due to CTins. This method was validated in three sample sets and applied to over 1600 patient samples. CTins caused C4A deficiency in more than 70% (76/105) of the carriers. Twenty per cent (76/381) of patients with a C4A deficiency would have been erroneously recorded as having none, if the CTins had not been assessed. C4A deficiency was more common in patients than a healthy reference population, (OR = 1.60, 95%CI = 1.02–2.52, p = 0.039). The number of functional C4 genes can be straightforwardly analyzed by real-time qPCR, also with SYBR® Green labelling. Determination of CTins increases the frequency of C4A deficiency and thus helps to elucidate the genotypic versus phenotypic disease associations. |
| Databáze: | OpenAIRE |
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