Real-time PCR quantification of human complement C4A and C4B genes

Agnes Szilagyi, B. Blaskó, Denes Szilassy, G. Füst, M. Sasvári, Z. Rónai

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

Background: The fourth component of human complement (C4), an essential factor of the innate immunity, is represented as two isoforms (C4A and C4B) in the genome. Although these genes differ only in 5 nucleotides, the encoded C4A and C4B proteins are functionally different. Based on phenotypic determination, unbalanced production of C4A and C4B is associated with several diseases, such as systemic lupus erythematosus, type 1 diabetes, several autoimmune diseases, moreover with higher morbidity and mortality of myocardial infarction and increased susceptibility for bacterial infections. Despite of this major clinical relevance, only low throughput, time and labor intensive methods have been used so far for the quantification of C4A and C4B genes. Results: A novel quantitative real-time PCR (qPCR) technique was developed for rapid and accurate quantification of the C4A and C4B genes applying a duplex, TaqMan based methodology. The reliable, single-step analysis provides the determination of the copy number of the C4A and C4B genes applying a wide range of DNA template concentration (0.3-300 ng genomic DNA). The developed qPCR was applied to determine C4A and C4B gene dosages in a healthy Hungarian population (N = 118). The obtained data were compared to the results of an earlier study of the same population. Moreover a set of 33 samples were analyzed by two independent methods. No significant difference was observed between the gene dosages determined by the employed techniques demonstrating the reliability of the novel qPCR methodology. A Microsoft Excel worksheet and a DOS executable are also provided for simple and automated evaluation of the measured data. Conclusions: This report describes a novel real-time PCR method for single-step quantification of C4A and C4B genes. The developed technique could facilitate studies investigating disease association of different C4 isotypes.

Original languageEnglish
Article number1
JournalBMC Genetics
Volume7
DOIs
Publication statusPublished - Jan 10 2006

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Real-Time Polymerase Chain Reaction
Gene Dosage
Genes
Complement C4
DNA
Type 1 Diabetes Mellitus
Bacterial Infections
Innate Immunity
Systemic Lupus Erythematosus
Population
Autoimmune Diseases
Protein Isoforms
Nucleotides
Myocardial Infarction
Genome
Morbidity
Mortality
Proteins

ASJC Scopus subject areas

  • Genetics
  • Genetics(clinical)

Cite this

Real-time PCR quantification of human complement C4A and C4B genes. / Szilagyi, Agnes; Blaskó, B.; Szilassy, Denes; Füst, G.; Sasvári, M.; Rónai, Z.

In: BMC Genetics, Vol. 7, 1, 10.01.2006.

Research output: Contribution to journalArticle

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