The SALSA MLPA
Probemixes P110 FCGR mix 1 and P111 FCGR mix 2 are research use only (RUO)
assays for the detection of genomic rearrangements and point mutations in the 180 kb FCGR2/3 locus at 1q23.3.
Receptors for the Fc portion of IgG play an essential role in the protection of the organism against foreign antigens by removing antigen-antibody complexes from the circulation. Receptors are present on monocytes, macrophages, neutrophils, natural killer (NK) cells and T and B lymphocytes. The receptors participate in diverse functions, such as phagocytosis of immune complexes and modulation of antibody production by B cells. Genes for several low-affinity Fc gamma receptors are clustered on chromosome 1q23.3 Within a 180 kb chromosomal area are genes for the FCGR2A, FCGR2B, FCGR2C, FCGR3A and FCGR3B proteins. In addition, this region contains genes for the HSPA6 and HSPA7 heat shock proteins.
Due to high similarity between these FCGR
genes and their close proximity, gene rearrangements are frequent in this chromosomal region. Various functionally relevant polymorphisms (SNPs) in these genes, as well as copy number variation of the FCGR2C
genes, have been reported. The MLPA probemixes P110/P111 FCGR cover the mentioned FCGR
genes and are intended to detect both copy number changes of these genes as well as frequent polymorphisms and point mutations, such as FCGR2B
p.His166Arg, and FCGR2A
p.Gln62Trp. Probes for FCGR3B
Human Neutrophil Antigen 1 (HNA1) alleles NA1, NA2 and SH, and FCGR2C
STOP, classic and non-classic ORF haplotypes are also included.
This SALSA MLPA Probemix is not CE/FDA registered for use in diagnostic procedures. Purchase of this product includes a limited license for research purposes.
The SALSA MLPA Probemix P110-C1 FCGR mix 1 contains a total of 40 MLPA probes with amplification products between 130 and 494 nucleotides (nt) (Table 1a). The SALSA MLPA Probemix P111-C1 FCGR mix 2 contains a total of 38 MLPA probes with amplification products between 130 and 490 nt (Table 1b).
Both P110 FCGR mix 1 and P111 FCGR mix 2 contain 14 probes for determination of copy number changes and genomic rearrangements (Table 2). P110 FCGR also contains 18, and P111 FCGR 16, mutation-specific probes which will only generate a signal when the mutation is present (Table 3). Both probemixes contain eight reference probes targeting copy number stable regions. Complete probe sequences and the identity of the genes detected by the reference probes are available online (www.mlpa.com
In addition, the probemixes contain nine quality control fragments generating amplification products between 64 and 105 nt: four DNA Quantity fragments (Q-fragments), two DNA Denaturation fragments (D-fragments), one Benchmark fragment, and one chromosome X and one chromosome Y-specific fragment. More information on how to interpret observations on these control fragments can be found in the MLPA General Protocol and online at www.mlpa.com
SALSA Reference Selection DNA SD038-S02:
The selection of suitable reference DNA samples that can be used with P110/P111 FCGR is complicated. To facilitate the selection of suitable reference DNA samples from your own sample collection, a reference selection DNA sample (catalogue number SD038) is provided with these probemixes from MRC-Holland. Reference selection DNA SD038-S02 should only be used for initial experiments on DNA samples from healthy individuals with the intention to identify suitable reference samples. SD038-S02 should not be used as a reference sample in subsequent experiments.
To aid in determining copy numbers of your own selected reference samples, the copy numbers of Reference Selection DNA SD038-S02 are detailed in the SD038-S02 product description, available online: www.mlpa.com
Sample DNA developed for this product: