As formulacoes CPS1
,5 e CPS3,0 foram as mais aceitas, enquanto a CPS4,5 apresentou menor aceitacao.
The target region encompassed the entire genomic regions of OTC, NAGS, and CPS1, along with 6 control loci for the assessment of enrichment efficacy (see Methods in the Data Supplement that accompanies the online version of this article at http://www.
m], and a lower tiling density than probes for OTC or CPS1 (see Table 3 in the online Data Supplement), suggesting that NAGS has sequence properties that are more challenging for MSC and NGS.
Furthermore, we observed that OTC had a lower median sequencing depth than CPS1 in sample CPSA.
A majority (94%) of these sequences mapped to a specific section of approximately 1800 bp in CPS1, a region for which no capture probes had been designed (Fig.
All OTC, NAGS, and CPS1 exons with flanking intronic and untranslated regions were screened for potential sequence variants, and parts of these screened regions were resequenced with Sanger technology (see Table 4 in the online Data Supplement).
Capturing the complete genomic regions of OTC, CPS1, and NAGS permitted the simultaneous and comprehensive screening for disease-causing mutations in 3 genes involved in inborn UCDs.
7] Human genes: CPS1, carbamoyl-phosphate synthetase 1, mitochondrial; NAGS, N-acetylglutamate synthase; OTC, ornithine carbamoyltransferase.
5] Human genes: DMD, dystrophin (muscular dystrophy, Duchenne and Becker types); GLDC glycine dehydrogenase (decarboxylating); SLC25A13, solute carrier family 25, member 13 (citrin); ARC811, ATP-binding cassette, sub-family B (MDR/TAP), member 11; ATP881, ATPase, class I, type 8B, member 1; OTC, ornithine carbamoyltransferase; CPS1
, carbamoyl-phosphate synthetase 1, mitochondrial; CY88, cytochrome b-245, beta polypeptide (chronic granulomatous disease); XK, X-linked Kx blood group (McLeod syndrome); RPGR, retinitis pigmentosa GTPase regulator; MECP2, methyl CpG binding protein 2 (Rett syndrome); A551, argininosuccinate synthetase 1; ASL, argininosuccinate lyase; ARC84, ATP-binding cassette, sub-family B (MDR/TAP), member 4.
The patient with CPS1 deficiency showed a heterozygous pattern with two peaks at 74.
m]s of the PCR products are large, they can be distinguished from each other using only SYBR Green I dye, as shown in the detection of the 9-bp deletion in CPS1 deficiency.
Name Role Position [alpha]-GAL 2Del-S PCR primer 10 928 (a) 2Del-AS PCR primer 11 114 2Del-F Probe 10 995 2Del-LC Probe 11 016 CPS1 9Del-S PCR primer 811 (b) 9Del-AS PCR primer 865 Name Sequence (5' [right arrow] 3') Modification [alpha]-GAL 2Del-S GGGCCACTTATCACTAGTTGC None 2Del-AS TGATGAAGCAGGCAGGAT None 2Del-F GTGGGAACGACCTCTCTCAG 3'-Fluorescein 2Del-LC CTTAGCCTGGGCTGTAGCTATGATA 5'-LC Red 640 3'-Phosphorylation CPS1 9Del-S GCAGAACCACTAATTCAG None 9Del-AS GCTCCTTGCGATCACTCT None (a,b) Positions of primers and probes at the 5' termini, according to GenBank accession number: (a) X14448 for [alpha]-GAL and (b) Y15793 for CPS1.