Standardisation of multiplex fluorescent short tandem repeat analysis for chimerism testing

Nollet, Friedel; Billiet, Johan; Selleslag, Dominik; Criel, A.

doi:10.1038/sj.bmt.1703162

Cited by 107 publications

(86 citation statements)

References 13 publications

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“…DNA extraction and sequencing in both parents revealed no mutation in CASR gene, and as molecular analysis using microsatellite markers (24) indicated that the paternity was as stated, a diagnosis of a de novo heterozygous CASR mutation was made. Figure 1 Clinical case 1.…”

Section: Casementioning

confidence: 99%

Mutations of calcium-sensing receptor gene: two novel mutations and overview of impact on calcium homeostasis

Livadariu¹,

Auriemma²,

Rydlewski³

et al. 2011

European Journal of Endocrinology

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Objective: Genetic disorders of calcium metabolism arise in a familial or sporadic setting. The calciumsensing receptor (CASR) plays a key role in maintaining calcium homeostasis and study of the CASR gene can be clinically useful in determining etiology and appropriate therapeutic approaches. We report two cases of novel CASR gene mutations that illustrate the varying clinical presentations and discuss these in terms of the current understanding of CASR function. Patients and methods: A 16-year-old patient had mild hypercalcemia associated with low-normal urinary calcium excretion and normal-to-high parathyroid hormone (PTH) levels. Because of negative family history, familial hypocalciuric hypercalcemia was originally excluded. The second patient was a 54-year-old man with symptomatic hypocalcemia, hyperphosphatemia, low PTH, and mild hypercalciuria. Familial investigation revealed the same phenotype in the patient's sister. The coding region of the CASR gene was sequenced in both probands and their available first-degree relatives. Results: The first patient had a novel heterozygous inactivating CASR mutation in exon 4, which predicted a p.A423K change; genetic analysis was negative in the parents. The second patient had a novel heterozygous activating CASR mutation in exon 6, which predicted a p.E556K change; the affected sister of the proband was also positive. Conclusions: We reported two novel heterozygous mutations of the CASR gene, an inactivating mutation in exon 4 and the first activating mutation reported to date in exon 6. These cases illustrate the importance of genetic testing of CASR gene to aid correct diagnosis and to assist in clinical management.

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Section: Casementioning

confidence: 99%

Mutations of calcium-sensing receptor gene: two novel mutations and overview of impact on calcium homeostasis

Livadariu¹,

Auriemma²,

Rydlewski³

et al. 2011

European Journal of Endocrinology

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“…Stutter bands are additional peaks, 2-6 base pairs shorter than the target allele and result from "slippery" amplification by the Taq polymerase during PCR (Nollet et al, 2001;Schraml and Lion, 2003). In the case of dinucleotide repeat loci, the most prevalent stutter band is generally two bases shorter than the main allele band.…”

Section: Discussionmentioning

confidence: 99%

“…Probably this short stutter band simply lacks one core repeat unit relative to the main band (Walsh et al, 1996). The amount of stutter bands produced is proportional to the amount of original allele and range between 0.5-13% (Nollet et al, 2001). However, the stutter band interference is highly variable among different markers and should be used as an important criterion to exclude a marker for this kind of assay.…”

Section: Discussionmentioning

confidence: 99%

Quantitative Fluorescent PCR (QF-PCR) on microsatellites, a fast and quantitative method to assay chimeric DNA samples in honeybees (Apis mellifera)

et al. 2006

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-The production of chimeras by nuclear transfer or cell transplantation requires functional tools for the detection of individuals that have successfully incorporated donor material. Quantitative Fluorescent PCR (QF-PCR) on microsatellites was employed on artificial chimeric honeybee DNA samples, to develop protocols for detection and quantification of donor genotypes after nuclear transfer and cell transplantations into honeybee embryos. Standard curves were produced for three microsatellite markers and demonstrate that there is a close to linear correlation between the amounts of donor genotypes estimated with QF-PCR and actual amounst of genotypes. The method was sensitive down to a 1% level and could detect and quantify small amounts of donor DNA in artificially mixed samples. QF-PCR offers fast and sensitive detection and quantification of genotypes and, thus, provides a useful tool for studies of chimeric honeybees and other species.Quantitative Fluorescent PCR (QF-PCR) / microsatellites / nuclear transfer / honeybee / Apis mellifera

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“…Peripheral blood samples were collected from the donor and recipient before SCT to establish a pre-transplant STR profile for each donor-recipient pair. These profiles were applied in the post-transplant period when donor chimerism was calculated as described by Nollet et al 6 The recipient was considered to achieve complete donor chimerism if 100% of peripheral blood or bone marrow cells were of donor origin.…”

Section: Hematologic Recovery and Engraftmentmentioning

confidence: 99%

Comparison of New Flu-Bu12-Tg Conditioning With the Standard Bu-Cy Myeloablative Regimen in Patients Undergoing Allogeneic Stem Cell Transplantation for Acute Myeloid Leukemia

Raida¹,

Tuček²,

Faber³

et al. 2011

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub

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Aims. This study compares the outcomes of patients with high-risk acute myeloid leukemia (AML) who underwent allogeneic stem cell transplantation (SCT) after conditioning combining busulfan (16 mg/kg orally) and cyclophosphamide (120 mg/kg intravenously) (BU-CY) with those allografted after administration of fludarabine (150 mg/m 2 intravenously), busulfan (12 mg/kg orally) and thymoglobulin (6 mg/kg intravenously) (FLU-BU12-TG).Material and methods. SCT after BU-CY and FLU-BU12-TG was performed in 21 and 10 AML patients. There were no significant differences between groups in number of patients treated in complete disease remission, gender, age, donors, CD34+, mononuclear cell (MNC) count in the graft and follow-up period. However, significantly more SCTs from unrelated (90% vs. 19%; p=0.00018) and HLA-mismatched donors (50% vs. 0%; p=0.0004) were performed in the FLU-BU12-TG group. The Cox proportional hazards model was used to assess the risk of post-transplant AML relapse and non-relapse mortality (NRM). The probability of post-transplant 2-year event-free survival (EFS) and overall survival (OS) were calculated using the Kaplan-Meier method.Results. No significant differences were found between the FLU-BU12-TG and BU-CY groups in risk of AML relapse (HR=1.036; 95% CI [0.102 -10.47]; p=0.9), post-transplant NRM (HR=0.25; 95% CI [0.031 -1.96]; p=0.18), 2-year EFS (89% vs. 43%; p=0.19) or OS (79% vs. 57%; p=0.23).Conclusion. These pilot results demonstrate the efficacy of the new FLU-BU12-TG conditioning regimen in patients allografted for high-risk AML. This conditioning might become an alternative approach in patients at high risk of severe post-transplant complications after the standard BU-CY myeloablative regimen.

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Standardisation of multiplex fluorescent short tandem repeat analysis for chimerism testing

Cited by 107 publications

References 13 publications

Mutations of calcium-sensing receptor gene: two novel mutations and overview of impact on calcium homeostasis

Mutations of calcium-sensing receptor gene: two novel mutations and overview of impact on calcium homeostasis

Quantitative Fluorescent PCR (QF-PCR) on microsatellites, a fast and quantitative method to assay chimeric DNA samples in honeybees (Apis mellifera)

Comparison of New Flu-Bu12-Tg Conditioning With the Standard Bu-Cy Myeloablative Regimen in Patients Undergoing Allogeneic Stem Cell Transplantation for Acute Myeloid Leukemia

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