Integration of Multiple PCR Amplifications and DNA Mutation Analyses by Using Oligonucleotide Microchip

OBJECTIVE:To evaluate two hypothetical screening strategies for identifying children with long QT syndrome (LQTS), a cause of sudden death in childhood. METHODS:Families with KVLQT1, HERG, or SCN5A genotypes provided electrocardiographic (ECG) data for this analysis. This is the first time such genotype-phenotype information has been available. Using the LQTS genotype, the distributions of QTc in affected and unaffected children were established and screening performance for various QTc cut off points were modelled. The detection rate for DNA mutation analysis was determined from published experience. RESULTS:The mean QTc (SD) was 0.484 seconds (0.031) in 117 affected children and 0.420 seconds (0.021) in 133 unaffected children. A QTc cut off of 0.50 seconds in a population of 1 million children would identify 61 of the 200 affected children, and 100 unaffected children. Estimates of testing costs for a screening programme in the newborn period would be $327 869/case detected and $2 222 000/death avoided. Although not presently available for routine use, DNA analysis could, theoretically, identify 100 of the 200 children with LQTS within the same population, along with an estimated 100 unaffected children. CONCLUSION:The only available screening test for LQTS is ECG measurement. If DNA technology becomes available for screening, unit costs must be very low to be competitive. There are multiple problems with screening for LQTS: only a minority of children will be detected, cost/death avoided is high, and pilot studies would need to be in place for 5–10 years to document efficacy.

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A Multiplex PCR Assay Mediated by Universal Primers for the Detection of Adulterated Meat in Mutton

Journal of Food Protection ◽

10.4315/0362-028x.jfp-18-302 ◽

2019 ◽

Vol 82 (2) ◽

pp. 325-330 ◽

Cited By ~ 1

Author(s):

WANWAN LIU ◽

XIAONAN WANG ◽

JING TAO ◽

BANGSHENG XI ◽

MAN XUE ◽

...

Keyword(s):

Multiplex Pcr ◽

Detection System ◽

Meat Products ◽

Pcr Primers ◽

Rapid Identification ◽

Food Samples ◽

Detection Sensitivity ◽

Universal Primers ◽

Multiplex Pcr Assay ◽

Multiple Pcr

ABSTRACT This study aimed to establish a multiplex PCR detection system mediated by “universal primers,” which would be able to determine whether mutton meat contained nonmutton ingredients from rats, foxes, and ducks. Based on the sequence variation of specific mitochondrial genes, nine different multiplex PCR primers were designed, and four kinds of meat products were rapidly identified by electrophoresis using an optimized multiplex PCR system based on the molecular weight differences of the amplified products. Multiplex PCR applications optimized for meat food source from food samples for testing was used to verify the accuracy of the identification method. The results showed that the primers in multiple PCR system mediated by universal primers could be used for the rapid identification of rat, fox, duck, and sheep meat in mutton products, and the detection sensitivity could reach 0.05 ng/μL. The identification of food samples validated the practical value of this method. Therefore, a multiplex PCR system mediated by universal primers was established, which can be used to quickly identify the origin of animal ingredients from rats, foxes, and ducks in mutton products.

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The mitochondrial DNA mutation T12297C affects a highly conserved nucleotide of tRNALeu(CUN) and is associated with dilated cardiomyopathy

European Journal of Human Genetics ◽

10.1038/sj.ejhg.5200622 ◽

2001 ◽

Vol 9 (4) ◽

pp. 311-315 ◽

Cited By ~ 32

Author(s):

Maurizia Grasso ◽

Marta Diegoli ◽

Agnese Brega ◽

Carlo Campana ◽

Luigi Tavazzi ◽

...

Keyword(s):

Mitochondrial Dna ◽

Dilated Cardiomyopathy ◽

Mitochondrial Dna Mutation ◽

Dna Mutation

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Intramyocellular lipid excess in the mitochondrial disorder MELAS

Neurology Genetics ◽

10.1212/nxg.0000000000000160 ◽

2017 ◽

Vol 3 (3) ◽

pp. e160 ◽

Cited By ~ 5

Author(s):

Sailaja Golla ◽

Jimin Ren ◽

Craig R. Malloy ◽

Juan M. Pascual

Keyword(s):

Mitochondrial Dna ◽

Fat Metabolism ◽

Mitochondrial Disorder ◽

Scientific Investigation ◽

Resonance Spectroscopy ◽

Lipid Deposition ◽

Metabolic Dysfunction ◽

Intramyocellular Lipid ◽

Mitochondrial Dna Mutation ◽

Dna Mutation

Objective:There is a paucity of objective, quantifiable indicators of mitochondrial disease available for clinical and scientific investigation.Methods:To this end, we explore intramyocellular lipid (IMCL) accumulation noninvasively by 7T magnetic resonance spectroscopy (MRS) as a reporter of metabolic dysfunction in MELAS (mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes). We reasoned that mitochondrial dysfunction may impair muscle fat metabolism, resulting in lipid deposition (as is sometimes observed in biopsies), and that MRS is well suited to quantify these lipids.Results:In 10 MELAS participants and relatives, IMCL abundance correlates with percent mitochondrial DNA mutation abundance and with disease severity.Conclusions:These results indicate that IMCL accumulation is a novel potential disease hallmark in MELAS.

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