The Clinical Diagnostics Service of the Molecular Otolaryngology & Renal Research Laboratories is a CLIA-approved, Joint Commission-accredited diagnostic laboratory that offers mutation screening of several genes.
CFH-CFHR5 Genomic Region
Genetic analyses for C3 glomerulopathy (C3G) and atypical hemolytic uremic syndrome (aHUS) must include suitable technologies to detect copy number variation, hybrid genes and other complex genomic rearrangments in the CFH/CFHRs genomic region (Goodship, TH et al., 2017). We interrogate this region using Multiplex Ligation-Dependant Probe Amplification.
Indications for screening
Screening is offered to persons with C3G and aHUS.
MORL screening methodology
Multiplex Ligation-Dependent Probe Amplification (MLPA) is used to detect non-allelic homologous recombination events within the CFH – CFHR5. This assay is based on sequence-specific hybridization and the subsequent ligation of two directly flanking half-probes on a target nucleic acid sequence. Only when these half-probes are ligated can the resultant fragment serve as a template for PCR amplification. Multiple probes are used to cross-check validity.
The standard deviation rates for each probe are 4%-10% (Schouten, J et al., 2002). MLPA has 95% confidence intervals for positive and negative predictive accuracies of 0.951-0.996 and 0.9996-1 respectively (Ahn JW et al., 2007). With the use of multiple probes, the likelihood of a spurious result is remote.
Turnaround time is approximately 1 month.
8 - 10 cc. whole blood in lavender (EDTA) top tubes OR 10 μg DNA, minimum concentration: 50 ng/μl (A260/A280 1.8-2) resuspended in 0.1mM EDTA (10mM Tris HC1, 0.1mM EDTA, pH 8, Teknova Cat#T0220).
Cost & CPT Codes
See the MORL Testing Menu
Kidneeds (not-for-profit foundation dedicated to the cure of DDD)
Foundation for Children with Atypical HUS
Goodship TH, Cook HT, Fakhouri F, Fervenza FC, Frémeaux-Bacchi V, Kavanagh D, Nester CM, Noris M, Pickering MC, Rodríguez de Córdoba S, Roumenina LT, Sethi S, Smith RJ; Conference Participants. Atypical hemolytic uremic syndrome and C3 glomerulopathy: conclusions from a "Kidney Disease: Improving Global Outcomes" (KDIGO) Controversies Conference. Kidney Int. 2016 Dec 15. pii: S0085-2538(16)30604-4. doi: 10.1016/j.kint.2016.10.005. [Epub ahead of print], 91(3):539-551, 2017.PubMed PMID: 27989322