LightCyder peR for the Poiymorphisms - 308 an d -238 in the TNF Alpha Gene IIiII
Results
Based on the GenBank sequences of the TNF-a and TNF-13 genes, we established
PCR protocols for genotyping analysis for three common single nucleotide polymorphisms (SNPs) in the TNF gene cluster (Figs. 1-3). Genotype assignments
were verified by sequencing on an ABI Prism 310 Sequencer (Applied Biosystems,
Rotkreuz, Switzerland).
Comments
The TNF-a G-23BA, TNF-a G-30BA and LT-a/TNF-13 Ncol polymorphisms (A329G)
are frequent genetic alterations that have been linked to a great variety of disease
states: various autoimmune diseases, atopy [2B] and asthma [13], an increased risk
of rejection after transplantation and the response to hepatitis Band C viruses [9,
10,22] and chlamydia trachomatis [6]. Particularly, TNF-a G-23BA appears to identify a small subgroup of severely poly traumatized patients likely to develop sepsis
and may be used as a prognostic tool [26,27].
As TNF is a major mediator in the pathogenesis of sepsis and its sequelae, individuals with a genetic determination for high TNF responses may be at high risk
for development of organ failure and death when challenged with severe infec22 -
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Fig.!. Melting curve analysis of TNF-a -308 polymorphism. The black (GG) and the green (AA) curves show the homozygous variants of wild type and mutation, respectively. The red curve shows the heterozygous carrier of the mutation. The
blue base line represents the buffer