NephrolithiasisNext includes 30 genes associated with both syndromic and non-syndromic nephrolithiasis and nephrocalcinosis, such as primary hyperoxaluria (types 1, 2, and 3), adenine phosphoribosyltransferase (APRT) deficiency, cystinuria, Dent disease, familial hypomagnesemia and hypercalciuria and nephrocalcinosis (FHHNC), distal renal tubular acidosis, and others.

Genomic deoxyribonucleic acid (gDNA) is isolated from the patient’s specimen using a standardized methodology and quantified. Sequence enrichment of the exome and adjacent intronic nucleotides is carried out by a bait-capture methodology using long biotinylated oligonucleotide probes followed by polymerase chain reaction (PCR) and Next-Generation sequencing, and the 30 genes undergo comprehensive analysis. Reportable small insertions and deletions, potentially homozygous variants, and single nucleotide variant calls not satisfying established confidence thresholds based on coverage and alternative read ratios are verified by Sanger sequencing. Gross deletion/duplication analysis is assessed for all genes using a custom pipeline based on coverage and/or breakpoint analysis from NGS data and is followed by a confirmatory orthogonal method, as needed.