scholarly journals Mitochondrial DNA Mutation Detection by Electrospray Mass Spectrometry

2007 ◽  
Vol 53 (2) ◽  
pp. 195-203 ◽  
Author(s):  
Yun Jiang ◽  
Thomas A Hall ◽  
Steven A Hofstadler ◽  
Robert K Naviaux
Keyword(s):  
Mass Spectrometry ◽  
Mitochondrial Dna ◽  
Rflp Analysis ◽  
Endocrine Disorders ◽  
Ion Cyclotron Resonance ◽  
Accurate Identification ◽  
Mtdna Mutations ◽  
Fticr Ms ◽  
Pcr Rflp ◽  
Mtdna Variants

Abstract Background: Mitochondrial DNA (mtDNA) mutations cause a large spectrum of clinically important neurodegenerative, neuromuscular, cardiovascular, and endocrine disorders. We describe the novel application of electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry (ESI-FTICR MS) to the rapid and accurate identification of pathogenic mtDNA variants. Methods: In a blinded study, we used ESI-FTICR MS to analyze 24 unrelated samples of total cellular DNA containing 12 mtDNA variants and compared the results with those obtained by conventional PCR-restriction fragment length polymorphism (PCR-RFLP) analysis and gel electrophoresis. Results: From the 24-sample blinded panel, we correctly identified 12 of the samples as bearing an mtDNA variant and found the remaining 12 samples to have no pathogenic variants. The correlation coefficient between the 2 methods for mtDNA variant detection was 1.0; there were no false positives or false negatives in this sample set. In addition, the ESI-FTICR method identified 4 single-nucleotide polymorphisms (SNP) that had previously been missed by standard PCR-RFLP analysis. Conclusions: ESI-FTICR MS is a rapid, sensitive, and accurate method for the identification and quantification of mtDNA mutations and SNPs.

Identification of Hake Species (MerlucciusGenus) Using Sequencing and PCR−RFLP Analysis of Mitochondrial DNA Control Region Sequences

2001 ◽  
Vol 49 (11) ◽  
pp. 5108-5114 ◽  
Author(s):  
Javier Quinteiro ◽  
Rodrigo Vidal ◽  
Mónica Izquierdo ◽  
Carmen G. Sotelo ◽  
María José Chapela ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Control Region ◽  
Rflp Analysis ◽  
Mitochondrial Dna Control Region ◽  
Control Region Sequences ◽  
Pcr Rflp

Abstract 17097: Non-Synonymous Mitochondrial DNA Variants Are Common in Myocardial Tissue of Heart Failure Patients

Circulation ◽  
2020 ◽  
Vol 142 (Suppl_3) ◽  
Author(s):  
Pappu Ananya ◽  
Michael Binder ◽  
Yang Wanjun ◽  
Rebecca McClellan ◽  
Brittney Murray ◽  
...  
Keyword(s):  
Heart Failure ◽  
Mitochondrial Dna ◽  
Nuclear Dna ◽  
Left Ventricular ◽  
Individual Risk ◽  
Myocardial Tissue ◽  
Mtdna Mutation ◽  
Mtdna Mutations ◽  
Ventricular Tissue ◽  
Mtdna Variants

Introduction: Mitochondrial heart disease due to pathogenic mitochondrial DNA (mtDNA) mutations can present as hypertrophic or dilated cardiomyopathy, ventricular arrhythmias and conduction disease. It is estimated that the mutation rate of mtDNA is 10 to 20-fold higher than that of nuclear DNA genes due to damage from reactive oxygen species released as byproducts during oxidative phosphorylation. When a new mtDNA mutation arises, it creates an intracellular heteroplasmic mixture of mutant and normal mtDNAs, called heteroplasmy. Heteroplasmy levels can vary in various tissues and examining mtDNA variants in blood may not be representative for the heart. The frequency of pathogenic mtDNA variants in myocardial tissues in unknown. Hypothesis: Human ventricular tissue may contain mtDNA mutations which can lead to alterations in mitochondrial function and increase individual risk for heart failure. Methods: Mitochondrial DNA was isolated from 61 left ventricular myocardial samples obtained from failing human hearts at the time of transplantation. mtDNA was sequenced with 23 primer pairs. In silico prediction of non-conservative missense variants was performed via PolyPhen-2. Heteroplasmy levels of variants predicted to be pathogenic were quantified using allele-specific ARMS-PCR. Results: We identified 21 mtDNA non-synonymous variants predicted to be pathogenic in 17 hearts. Notably, one heart contained four pathogenic mtDNA variants (ATP6: p.M104; ND5: p.P265S; ND4: p.N390S and p.L445F). Heteroplasmy levels exceeded 90% for all four variants in myocardial tissue and were significantly lower in blood. No pathogenic mtDNA variants were identified in 44 hearts. Hearts with mtDNA mutations had higher levels of myocardial GDF-15 (growth differentiation factor-15; 6.2±2.3 vs. 1.3±0.18, p=0.045), an established serum biomarker in various mitochondrial diseases. Conclusions: Non-synonymous mtDNA variants predicted to be pathogenic are common in human left ventricular tissue and may be an important modifier of the heart failure phenotype. Future studies are necessary to correlate myocardial mtDNA mutations with cardiovascular outcomes and to assess whether serum GDF-15 allows identifying patients with myocardial mtDNA mutations.

scholarly journals Case Report: Identification of a Novel Variant (m.8909T>C) of Human Mitochondrial ATP6 Gene and Its Functional Consequences on Yeast ATP Synthase

Life ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 215
Author(s):  
Qiuju Ding ◽  
Róża Kucharczyk ◽  
Weiwei Zhao ◽  
Alain Dautant ◽  
Shutian Xu ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Atp Synthase ◽  
Atp Synthesis ◽  
Single Individual ◽  
Loss Of Function ◽  
Mtdna Mutations ◽  
Atp6 Gene ◽  
Novel Variant ◽  
Functional Consequences ◽  
Mtdna Variants

With the advent of next generation sequencing, the list of mitochondrial DNA (mtDNA) mutations identified in patients rapidly and continuously expands. They are frequently found in a limited number of cases, sometimes a single individual (as with the case herein reported) and in heterogeneous genetic backgrounds (heteroplasmy), which makes it difficult to conclude about their pathogenicity and functional consequences. As an organism amenable to mitochondrial DNA manipulation, able to survive by fermentation to loss-of-function mtDNA mutations, and where heteroplasmy is unstable, Saccharomyces cerevisiae is an excellent model for investigating novel human mtDNA variants, in isolation and in a controlled genetic context. We herein report the identification of a novel variant in mitochondrial ATP6 gene, m.8909T>C. It was found in combination with the well-known pathogenic m.3243A>G mutation in mt-tRNALeu. We show that an equivalent of the m.8909T>C mutation compromises yeast adenosine tri-phosphate (ATP) synthase assembly/stability and reduces the rate of mitochondrial ATP synthesis by 20–30% compared to wild type yeast. Other previously reported ATP6 mutations with a well-established pathogenicity (like m.8993T>C and m.9176T>C) were shown to have similar effects on yeast ATP synthase. It can be inferred that alone the m.8909T>C variant has the potential to compromise human health.

Timber species identification from chemical fingerprints using direct analysis in real time (DART) coupled to Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS): comparison of wood samples subjected to different treatments

Holzforschung ◽  
2019 ◽  
Vol 73 (11) ◽  
pp. 975-985 ◽  
Author(s):  
Maomao Zhang ◽  
Guangjie Zhao ◽  
Juan Guo ◽  
Alex C. Wiedenhoeft ◽  
Charles C. Liu ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Fourier Transform ◽  
Statistical Analysis ◽  
Real Time ◽  
Cyclotron Resonance ◽  
Direct Analysis ◽  
Species Level ◽  
Ion Cyclotron Resonance ◽  
Timber Species ◽  
Fticr Ms

Abstract Timber genus identification based on the anatomical features of wood is well established in botany. However, species-level wood identification is not always possible based on traditional wood morphology techniques alone. To compensate for the deficiencies of traditional methods, direct analysis in real time coupled to Fourier transform ion cyclotron resonance mass spectrometry (DART-FTICR-MS) was used to obtain the mass spectral fingerprints of different timber species. Using heartwood samples of two morphologically similar species, Pterocarpus santalinus and Pterocarpus tinctorius, subjected to different treatments, i.e. solvent extractions and powdered samples as well as air-dried samples and samples dried at low and high temperatures, we observed distinct chemical signatures for the wood samples from the two species, enabling rapid species-level identification when multivariate statistical analysis was adopted. The supervised orthogonal partial least squares discriminant analysis (OPLS-DA) models for samples subjected to different treatments all exhibited accurate differentiation performance of the explained fraction of variance of classes (R2Y = 0.936–0.987) and the cross-validated fraction of variance of classes (Q2 = 0.857–0.949). Compared with solvent types and the physical form of the sample, the drying treatment method had a greater impact on the chemical fingerprint from DART-FTICR-MS. Air-dried wood chips were the optimal samples for the DART-FTICR-MS method coupled with statistical analysis.

PCR-based genetic identification of marlin and other billfish

1998 ◽  
Vol 49 (5) ◽  
pp. 383 ◽  
Author(s):  
B. H. Innes ◽  
P. M. Grewe ◽  
R. D. Ward
Keyword(s):  
Mitochondrial Dna ◽  
Genetic Test ◽  
Rflp Analysis ◽  
Restriction Enzymes ◽  
Genetic Identification ◽  
Dna Molecule ◽  
Pcr Rflp ◽  
D Loop ◽  
Specific Variation ◽  
Species Specific

A genetic test was developed for the identification of the six species of billfish found in Australian waters (black marlin, Indo–Pacific blue marlin, striped marlin, Indo–Pacific sailfish, shortbill spearfish and broadbill swordfish). The test was based on the PCR–RFLP analysis of a 1400 bp region of the mitochondrial DNA molecule, the d-loop, using four restriction enzymes (Hinf I, Rsa I and Sau3A I andTaq I). A total of 33 composite haplotypes were observed among 160 fish; all were species-specific. Three of the species—black marlin, striped marlin and broadbill swordfish—showed sufficient intra-specific variation to be useful in population structure analyses.

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