Search results for "Genotyping"

ABO genotyping by PCR-RFLP and cloning and sequencing

2005

A refined PCR-RFLP based method was established to genotype ABO blood groups. The main objective of this study was to make the techniques also suitable for working with degraded DNA. Specific primer design was carried out to choose fragments shorter than 200 bp as necessary in forensic and archaeological applications. Four fragments of exon 6 and 7 of the ABO gene were amplified and digested by in total 7 restriction endonucleases. Particular attention was paid to the base changes at nucleotide positions 261(delG), 297, 526, 703, 721, 771, 796 and 1060(delC) in order to distinguish the six common alleles A101, A201, B, O01, O02 and O03. Furthermore, this method also enables determination of…

Nucleotide variation in the VP7 gene affects PCR genotyping of G9 rotaviruses identified in Italy

2003

A modified (aFT9m) and a degenerate (aFT9d) version of the rotavirus G9-specific primer (aFT9) allowed strains that were previously untypable, because of point mutations accumulating at the primer binding site, to be G typed by reverse transcription-polymerase chain reaction. The strains were collected during 2001-2002 in Italy in hospitals of the Apulia region, from children affected by severe rotavirus-associated enteritis. Using a wide selection of G9 rotaviruses detected worldwide, sequencing of the G9 untypable strains, sequence comparison, and phylogenetic analysis showed that the Italian strains have strong genetic similarity (< or =99.4%) to G9 rotaviruses identified recently in man…

G2 rotavirus infections in an infantile population of the South of Italy: variability of viral strains over time.

2005

Rotavirus positive samples collected in Palermo, Italy, during 2002–2004 did not react with the G2 type-specific RV5:3 monoclonal antibodies (MAbs) and could be identified as G2 only by RT-PCR genotyping. The genetic variation of VP7 and VP4 antigenic proteins was studied in 14 G2 samples including a selection of both those successfully characterized by serotyping and those failing to be serotyped. The phylogenetic analysis performed on partial VP7 sequences showed a temporal clustering of these strains, with those isolated in Palermo in 2003 belonging to the same lineage of G2 MAbs-unreactive strains identified in UK in 1996–1997 and in Bari, Italy, in 2003–2004. A single amino acid substi…

APPROACHES TO STUDY BIODIVERSITY AND SALINITY TOLERANCE MECHANISMS IN MEDITERRANEAN FRUIT TREE SPECIES

Changes in global climate are going to increase the problem of drought and soil salinity in the Mediterranean area, where increases in temperature, drought and soil and water salinity are expected. Identification of plant genotypes with positive agronomic traits, such as salt tolerance, may reduce the effects of salinity and drought on productivity. Therefore, there is considerable effort being directed toward the development of salt-tolerant genotypes through plant breeding, with the aim to the introgression of salt-tolerance traits into new cultivars. The results presented in this thesis contributed to the develop of new tools to improve knowledge on the biodiversity and salinity toleranc…

Metodo per la tracciabilità/autenticazione di prodotti lattiero-caseari mono-razza

2016

Il brevetto consiste in un metodo per l’autenticazione e la tracciabilità genetica delle produzioni lattiero-casearie mono-razza su base molecolare con approccio statistico-probabilistico. Il metodo si basa sull’impiego di tecniche di analisi ad elevata processività (in particolare SNPs genotyping su Bead Chip Illumina) su pool di DNA e sull’applicazione di metodi computazionali innovativi. Il metodo è già stato validato su produzioni lattiero-casearie ovine quali la Vastedda della Valle del Belice (DOP), sia su prodotti lattiero caseari a DOP ottenuti in laboratorio, che su prodotti a DOP reperibili sul mercato. Il metodo può comunque essere esteso, su richiesta, a qualsiasi altro prodotto…

Analysis of Illumina BovineSNP50 BeadChip data in different Italian cattle breeds to identify useful markers for breed authentication of dairy and be…

2013

The identification of the breed of origin of cattle derived products has recently assumed particular relevance since the increasing interest in marketing mono-breed labelled cheese and beef. These products are usually sold at a higher price creating financial incentives to substituting them with lower value products coming from undifferentiated or common breeds or crossbred animals. Frauds can damage these valuable niche markets that, indirectly, may help to preserve genetic diversity through the economic incentive on raising local breeds, derived by the high value products, that are usually less productive. DNA analysis can help to identify fraudulent behaviour as we already demonstrated f…

Age-associated changes in circulatory fatty acids: new insights on adults and long-lived individuals

2022

Long-lived individuals (LLIs) are considered an ideal model to study healthy human aging. Blood fatty acid (FA) profile of a cohort of LLIs (90–111 years old, n = 49) from Sicily was compared to adults (18–64 years old, n = 69) and older adults (65–89 years old, n = 54) from the same area. Genetic variants in key enzymes related to FA biosynthesis and metabolism were also genotyped to investigate a potential genetic predisposition in determining the FA profile. Gas chromatography was employed to determine the FA profile, and genotyping was performed using high-resolution melt (HRM) analysis. Blood levels of total polyunsaturated FA (PUFA) and total trans-FA decreased with age, while the lev…

Genotyping of GII.4 and GIIb norovirus RT-PCR amplicons by RFLP analysis

2007

GII.4 and GIIb/Hilversum norovirus (NoV) strains appear to have a prominent epidemiological role in outbreaks or sporadic cases of human gastroenteritis. Sequence analysis, although laborious, is the reference method used for characterization of noroviruses. In this study a screening test is proposed to characterize GIIb and GII.4 NoVs based on restriction fragment length polymorphism (RFLP) analysis of amplicons obtained from the RNA-dependent RNA polymerase (RdRp) region. Virtual analysis of 793 RdRp sequences of GGI and GGII NoVs, retrieved from GenBank, and representative of global geographical origins on a long-time period, permitted the selection of four restriction enzymes, XmnI, Ahd…

Detection of the norovirus variants GGII.4 hunter and GGIIb/hilversum in Italian children with gastroenteritis.

2006

Noroviruses (NoVs) are important enteric pathogens of humans. Although they exhibit an impressive genetic diversity, few NoV strains appear to predominate worldwide. Limited epidemiological data are available on NoV gastroenteritis in Italy. In this study, we assessed the prevalence of human NoV in Italian children with gastroenteritis by using a reverse-transcription nested polymerase chain reaction (RT-PCR) assay specific for the RNA-dependent RNA polymerase (RdRp) on faecal samples collected throughout the 2004 surveillance activity in Palermo, Italy. NoVs were detected in 47% of the stool samples obtained from children <5 years age, admitted to hospital with acute non-bacterial gastroen…

Rotavirus umani ed animali di genotipo G3 responsabili di gastroenterite infantile a Palermo nel 1993-2005.

2008

I rotavirus di gruppo A sono i più frequenti agenti eziologici di gastroenterite virale sia nell’uomo che in numerose specie animali. Per valutare l’evoluzione di tali virus si è rivelato utile effettuare l’analisi di sequenza dei geni codificanti per le proteine del capside esterno VP7 e VP4, per la proteina del capside interno VP6 e per la proteina non strutturale NSP4. Attualmente sono stati descritti in natura: 15 genotipi G in base a VP7, 27 genotipi P in base a VP4, 4 sottogruppi VP6 e 5 genotipi NSP4. I rotavirus umani appartengono più frequentemente ai genotipi G1P[8], G3P[8] e G4P[8], che si associano con il SGII (VP6 correlato) e NSP4B, ovvero al genotipo G2P[4], associato a SGII …