QMPSF is sensitive and specific in the detection of NPHP1 heterozygous deletions
-
Eszter Jávorszky
,
Vincent Morinière
Abstract
Background:
Nephronophthisis, an autosomal recessive nephropathy, is responsible for 10% of childhood chronic renal failure. The deletion of its major gene, NPHP1, with a minor allele frequency of 0.24% in the general population, is the most common mutation leading to a monogenic form of childhood chronic renal failure. It is challenging to detect it in the heterozygous state. We aimed to evaluate the sensitivity and the specificity of the quantitative multiplex PCR of short fluorescent fragments (QMPSF) in its detection.
Methods:
After setting up the protocol of QMPSF, we validated it on 39 individuals diagnosed by multiplex ligation-dependent probe amplification (MLPA) with normal NPHP1 copy number (n=17), with heterozygous deletion (n=13, seven parents and six patients), or with homozygous deletion (n=9). To assess the rate of the deletions that arise from independent events, deleted alleles were haplotyped.
Results:
The results of QMPSF and MLPA correlated perfectly in the identification of 76 heterozygously deleted and 56 homozygously deleted exons. The inter-experimental variability of the dosage quotient obtained by QMPSF was low: control, 1.05 (median; range, 0.86−1.33, n = 102 exons); heterozygous deletion, 0.51 (0.42−0.67, n = 76 exons); homozygous deletion, 0 (0−0, n = 56 exons). All patients harboring a heterozygous deletion were found to carry a hemizygous mutation. At least 15 out of 18 deletions appeared on different haplotypes and one deletion appeared de novo.
Conclusions:
The cost- and time-effective QMPSF has a 100% sensitivity and specificity in the detection of NPHP1 deletion. The potential de novo appearance of NPHP1 deletions makes its segregation analysis highly recommended in clinical practice.
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
Research funding: Financial support for this work was provided by MTA-SE Lendulet Research Grant (LP2015-11/2015) of the Hungarian Academy of Sciences, OTKA K109718 and by the French-Hungarian bilateral project (PHC 34501SM Balaton, Hungarian Grant No. TeT_14_1-2015-0020).
Employment or leadership: None declared.
Honorarium: None declared.
Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.
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©2017 Walter de Gruyter GmbH, Berlin/Boston
Artikel in diesem Heft
- Frontmatter
- Editorial
- Biotin interference on immunoassay methods: sporadic cases or hidden epidemic?
- Reviews
- False biochemical diagnosis of hyperthyroidism in streptavidin-biotin-based immunoassays: the problem of biotin intake and related interferences
- Vitamin K plasma levels determination in human health
- Mini Review
- Point-of-care testing INR: an overview
- Opinion Paper
- Could accreditation bodies facilitate the implementation of medical guidelines in laboratories?
- Genetics and Molecular Diagnostics
- QMPSF is sensitive and specific in the detection of NPHP1 heterozygous deletions
- General Clinical Chemistry and Laboratory Medicine
- High-dose biotin therapy leading to false biochemical endocrine profiles: validation of a simple method to overcome biotin interference
- Multicenter performance evaluation of a second generation cortisol assay
- A rapid UPLC-MS/MS assay for the simultaneous measurement of fluconazole, voriconazole, posaconazole, itraconazole, and hydroxyitraconazole concentrations in serum
- A microplate assay to measure classical and alternative complement activity
- Serological diagnosis and prognosis of severe acute pancreatitis by analysis of serum glycoprotein 2
- Retrospective evaluation of the clinical utility of serological biomarkers in Chinese patients with inflammatory bowel disease: 2-year clinical experience
- Infrared analysis of lipoproteins in the detection of alcohol biomarkers
- Coexistence of anti-β2-glycoprotein I domain I and anti-phosphatidylserine/prothrombin antibodies suggests strong thrombotic risk
- Antiphosphatidylserine/prothrombin antibodies as biomarkers to identify severe primary antiphospholipid syndrome
- Cardiovascular Diseases
- The impact of admission neutrophil-to-platelet ratio on in-hospital and long-term mortality in patients with infective endocarditis
- Letters to the Editor
- Biotin interference in immunoassays mimicking subclinical Graves’ disease and hyperestrogenism: a case series
- A simple method to detect biotin interference on immunoassays
- Proposed classification for a variant of Kounis syndrome
- Lyophilized hemoglobin E control material for the dichlorophenol-indophenol (DCIP) test
- Procalcitonin variation before and after 100-km ultramarathon
- Preliminary study in specific activity of molecular components in allergy: implications for diagnostics and relationship with disease severity
- A 2-min at 4500 g rather than a 15-min at 2200 g centrifugation does not impact the reliability of 10 critical coagulation assays
- Immunoassay interference caused by heterophilic antibodies interacting with biotin
- Red blood cell distribution width and mean platelet volume are potential prognostic indices for patients with primary biliary cirrhosis
- Neutrophil CD64 molecule expression can predict bloodstream infection in septic shock patients
Artikel in diesem Heft
- Frontmatter
- Editorial
- Biotin interference on immunoassay methods: sporadic cases or hidden epidemic?
- Reviews
- False biochemical diagnosis of hyperthyroidism in streptavidin-biotin-based immunoassays: the problem of biotin intake and related interferences
- Vitamin K plasma levels determination in human health
- Mini Review
- Point-of-care testing INR: an overview
- Opinion Paper
- Could accreditation bodies facilitate the implementation of medical guidelines in laboratories?
- Genetics and Molecular Diagnostics
- QMPSF is sensitive and specific in the detection of NPHP1 heterozygous deletions
- General Clinical Chemistry and Laboratory Medicine
- High-dose biotin therapy leading to false biochemical endocrine profiles: validation of a simple method to overcome biotin interference
- Multicenter performance evaluation of a second generation cortisol assay
- A rapid UPLC-MS/MS assay for the simultaneous measurement of fluconazole, voriconazole, posaconazole, itraconazole, and hydroxyitraconazole concentrations in serum
- A microplate assay to measure classical and alternative complement activity
- Serological diagnosis and prognosis of severe acute pancreatitis by analysis of serum glycoprotein 2
- Retrospective evaluation of the clinical utility of serological biomarkers in Chinese patients with inflammatory bowel disease: 2-year clinical experience
- Infrared analysis of lipoproteins in the detection of alcohol biomarkers
- Coexistence of anti-β2-glycoprotein I domain I and anti-phosphatidylserine/prothrombin antibodies suggests strong thrombotic risk
- Antiphosphatidylserine/prothrombin antibodies as biomarkers to identify severe primary antiphospholipid syndrome
- Cardiovascular Diseases
- The impact of admission neutrophil-to-platelet ratio on in-hospital and long-term mortality in patients with infective endocarditis
- Letters to the Editor
- Biotin interference in immunoassays mimicking subclinical Graves’ disease and hyperestrogenism: a case series
- A simple method to detect biotin interference on immunoassays
- Proposed classification for a variant of Kounis syndrome
- Lyophilized hemoglobin E control material for the dichlorophenol-indophenol (DCIP) test
- Procalcitonin variation before and after 100-km ultramarathon
- Preliminary study in specific activity of molecular components in allergy: implications for diagnostics and relationship with disease severity
- A 2-min at 4500 g rather than a 15-min at 2200 g centrifugation does not impact the reliability of 10 critical coagulation assays
- Immunoassay interference caused by heterophilic antibodies interacting with biotin
- Red blood cell distribution width and mean platelet volume are potential prognostic indices for patients with primary biliary cirrhosis
- Neutrophil CD64 molecule expression can predict bloodstream infection in septic shock patients
