Search results for "Quantitative PCR"

Comparative multiplex dosage analysis in spinocerebellar ataxia type 2 patients.

2013

We developed a new application of comparative multiplex dosage analysis (CMDA) for evaluation of the ataxin 2 gene. Expansions of the triplet CAG can cause spinocerebellar ataxia type 2 (SCA2), a neurodegenerative disease with an autosomal-dominant mode of inheritance. Molecular diagnosis of SCA2 is routinely based on the use of conventional PCR to detect the CAG expansion. However, PCR does not amplify an allele with an expansion of many triplets (>80), which is typically found in infantile and juvenile forms of SCA2, thus leading to false negatives. We propose the analysis of the ATXN2 gene by CMDA to complement existing methods currently used for the detection of large expansions of the …

The experience of the International Consortium on Acute Promyelocytic Leukemia in monitoring minimal residual disease in acute promyelocytic leukaemia

2016

Molecular diagnostic tools for the detection and characterization of Phoma tracheiphila

2011

In the recent years PCR-based techniques for the identification and detection of Phoma tracheiphila, the causal agent of citrus mal secco disease, have been evaluated aiming to provide tools for biological and epidemiological studies. A wide collection of P. tracheiphila strains was used to evaluate and validate diagnostic protocols and a fAFLP method for fungal characterization. Conventional and real-time PCR protocols were successfully tested for the specific identification of P. tracheiphila and its detection in planta. A further improvement of the real-time PCR protocol and the DNA extraction methods allowed the quantification of the fungus both from naturally infected and artificially …

Increased serum miR-193a-5p during non-alcoholic fatty liver disease progression: Diagnostic and mechanistic relevance

2022

Background & Aims Serum microRNA (miRNA) levels are known to change in non-alcoholic fatty liver disease (NAFLD) and may serve as useful biomarkers. This study aimed to profile miRNAs comprehensively at all NAFLD stages. Methods We profiled 2,083 serum miRNAs in a discovery cohort (183 cases with NAFLD representing the complete NAFLD spectrum and 10 population controls). miRNA libraries generated by HTG EdgeSeq were sequenced by Illumina NextSeq. Selected serum miRNAs were profiled in 372 additional cases with NAFLD and 15 population controls by quantitative reverse transcriptase PCR. Results Levels of 275 miRNAs differed between cases and population controls. Fewer differences were seen wi…

Determinants of the distribution of nitrogen-cycling microbial communities at the landscape-scale

2010

Little information is available regarding the landscape-scale distribution of microbial communities and its environmental determinants. However, a landscape perspective is needed to understand the relative importance of local and regional factors and land management for the microbial communities. In this manuscript, we investigated the distribution of functional microbial communities involved in N-cycling and of the total bacterial and crenarchaeal communities over 107 sites using a grid with a 16 km lag distance within Burgundy, a 31 500 km2 region in France. After quantifying the abundances of the total bacterial, crenarchaeal, nitrate-reducing, denitrifying and ammonia-oxidizing communit…

Technosols to reclaim industrial wastelands: depth distribution of abundance and activity of N-cycling microbial communities

2012

International audience; Construction of Technosols through assemblage of treated soil and recycled wastes is an innovative option for the restoration of degraded lands and re-use of industrial wastes. Recent studies have evidenced that Technosols could support soil functions such as primary production but the knowledge about other ecosystemic services, such as nutrient cycling, is limited. In this work, we investigated the abundance and the activity of microbial communities involved in N-cycling in different horizons (0–15, 15–35, 35–70 cm) of two types of Technosols constructed to reclaim an industrial wasteland. The estimation by real-time PCR of the abundances of the different microbial …

Study of the aminopeptidase N gene family in the lepidopterans Ostrinia nubilalis (Hübner) and Bombyx mori (L.): Sequences, mapping and expression

2010

Aminopeptidases N (APNs) are a class of ectoenzymes present in lepidopteran larvae midguts, involved in the Bacillus thuringiensis (Bt) toxins mode of action. In the present work, seven aminopeptidases have been cloned from the midgut of Ostrinia nubilalis, the major Lepidopteran corn pest in the temperate climates. Six sequences were identified as APNs because of the presence of the HEXXH(X)18E and GAMEN motifs, as well as the signal peptide and the GPI-anchor sequences. The remaining sequence did not contain the two cellular targeting signals, indicating it belonged to the puromycin-sensitive aminopeptidase (PSA) family. An in silico analysis allowed us to find orthologous sequences in Bo…

Glucose transporter 4 mRNA expression in subcutaneous adipose tissue of women with PCOS remains unchanged despite metformin withdrawal: is there a ce…

2021

Purpose: Metformin induces GLUT-4 mRNA expression in insulin target tissues in PCOS. It is unclear how long this impact is sustained after withdrawal of metformin. We aimed to compare the effect of metformin withdrawal on GLUT-4 mRNA expression in subcutaneous adipose tissue after prior short (ST, 1 year, N = 11) and long term (LT, at least 3 years, N = 13) treatment in obese PCOS women. Methods: At baseline and 6 months after withdrawal, biopsy of subcutaneous adipose tissue followed by quantitative PCR analysis was performed to determine GLUT-4 mRNA expression. Results: We found no time/effect differences in GLUT-4 mRNA expression in ST (2-dCt at baseline 0.42 (0.16–0.48) vs 2-dCt after 6…