PubMed Health⌕ Search

Biomedical subjects

Matteo Adinolfi

Publications and source records attributed to Matteo Adinolfi.

10 recordsLinked to original sources
PubMed2006

Rapid prenatal diagnosis by QF-PCR: evaluation of 30,000 consecutive clinical samples and future applications.

Rapid prenatal diagnoses of major chromosome abnormalities can be performed on a large scale using highly polymorphic short tandem repeats (STRs) amplified by the quantitative fluorescent polymerase chain reaction (QF-PCR). The assay was introduced as a preliminary investigation to remove the anxiety of the parents waiting for the results by conventional cytogenetic analysis using amniotic fluid or chorionic cells. However, recent studies, on the basis of the analyses of several thousand samples, have shown that this rapid approach has a very high rate of success and could reduce the need for cytogenetic investigations. Its high efficiency, for example, allows early interruption of affected fetuses without the need of waiting for completion of fetal karyotype. The main advantages of the QF-PCR are its accuracy, speed, automation, and low cost that allows very large number of samples to be analyzed by few operators. Here, we report the results of using QF-PCR in a large series of consecutive clinical cases and discuss the possibility that, in a near future, it may even replace conventional cytogenetic analyses on selected samples.

Cytogenetic Analysis↗
PubMed2005

Versatile use of ytterbium(III) triflate and acid washed molecular sieves in the activation of glycosyl trifluoroacetimidate donors. Assemblage of a biologically relevant tetrasaccharide sequence of Globo H.

The nonreducing tetrasaccharide terminus of Globo H has been assembled in good yield and excellent stereocontrol exclusively by using mild and moisture stable agents such as Yb(OTf)(3) and acid washed molecular sieves for the activation of glycosyltrifluoroacetimidate donors in the glycosylation steps.

Antigens, Tumor-Associated, Carbohydrate↗
PubMed2005

Non-invasive screening and rapid QF-PCR assay can greatly reduce the need for conventional cytogenetic analyses in prenatal diagnosis.

In 2004, the UK National Screening Committee suggested that rapid screening tests, such as fluorescence in-situ hybridization (FISH) and/or quantitative fluorescence PCR (QF-PCR), should replace prenatal diagnosis of Down syndrome performed by conventional karyotyping. However, doubts have been expressed that replacement of conventional cytogenetic investigations would result in a substantial number of infants affected by preventable handicaps. Based on a brief analysis of 28,000 prenatal tests performed in genetic units, this paper discusses the advantages of using QF-PCR. All normal fetuses were correctly diagnosed without false positive results and approximately 93% major chromosome disorders were detected by the molecular approach. The need for cytogenetic tests was thus greatly reduced, since pregnancy can be terminated, if necessary, without the need to confirm the results. A careful combination of accurately performed non-invasive ultrasound and maternal blood tests, eventually followed by QF-PCR, should reduce the need for conventional chromosome analyses.

Chromosome Aberrations↗
PubMed2004

Modulating the activity of oligonucleotides by carbohydrate conjugation: solid phase synthesis of sucrose-oligonucleotide hybrids.

In order to expand the repertoire of available oligosaccharide-oligonucleotide hybrids, the on-line solid phase synthesis of oligonucleotides conjugated at the 3'-and/or 5'-end with a preformed disaccharide unit has been performed. The key compound in the synthetic scheme described here is an appropriate phosphoramidite derivative of fully protected sucrose, used in association with a solid support functionalized with DMT-protected sucrose. The sucrose units at both ends of selected oligonucleotide sequences were shown to increase their chemical and enzymatic stability, while not interfering with duplex formation and with the ability of G-rich sequences to adopt a quadruplex structure.

Chromatography, High Pressure Liquid↗
PubMed2003

Activation of glycoyl trihaloacetimidates with acid-washed molecular sieves in the glycosidation reaction.

[reaction: see text] Commercially available 4 A acid washed molecular sieves (4 A AW 300 MS) are efficient activators of glycosyl trichloro- and N-phenyltrifluoroacetimidates. These promoters allow glycosidation of primary and secondary saccharidic acceptors to be performed in high yield, under very mild conditions and by an experimentally simple procedure. In addition, the recyclability of such promoters also has been demonstrated.

Acids↗
PubMed2003

HLA-G positive trophoblastic cells in transcervical samples and their isolation and analysis by laser microdissection and QF-PCR.

OBJECTIVE: To assess the frequency of cytotrophoblastic cells in endocervical samples collected by lavage at early stages of gestation using a specific anti-HLA-G McAb (G233). From a set of four selected samples, cells identified by immunostaining were collected by laser microdissection and then tested by quantitative fluorescent polymerase chain reaction (QF-PCR) for the presence of paternally derived DNA markers, in order to establish their fetal origin. METHODS: Syncytial fragments and cytotrophoblastic cells from 23 transcervical samples were identified by immunostaining with McAb G233 reacting against HLA-G antigen and with antibodies against cytokeratin. Slides from the same samples were also tested by fluorescent in situ hybridization (FISH), while selected samples were analysed by QF-PCR. Slides from four samples retrieved from mothers with male fetuses were immunolabelled and then cytotrophoblastic cells, syncytial fragments and maternal epithelial cells were collected by laser microdissection and tested by QF-PCR. RESULTS: All endocervical samples retrieved from mothers with male fetuses were found to contain some cells with chromosome Y-specific signals when tested by FISH. Using McAb anti- HLA-G, cytotrophoblastic cellular elements were detected in about 50% of the samples. From four samples, cellular elements identified by immunostaining as cytotrophoblast or syncytial fragments were collected by laser microdissection and shown to be of fetal origin when tested by QF-PCR for the presence of fetal DNA markers. CONCLUSIONS: These results confirm that, during an early phase of gestation, fetal cells are released in the lower uterine cavity and that they can be isolated and analysed for prenatal diagnosis of single gene defects and aneuploidies.

Adult↗
PubMed2003

Comparison between fluorescence in situ hybridization (FISH) and quantitative-fluorescent polymerase chain reaction (QF-PCR) for the detection of aneuploidies in single blastomeres.

OBJECTIVES: The aim of our investigation was to compare the efficiencies of the fluorescence in situ hybridization (FISH) and the quantitative-fluorescent PCR (QF-PCR) methods for the detection of sexing and numerical chromosome disorders in single blastomeres collected from the same preimplantation human embryos. METHODS: FISH analysis was carried out on 145 blastomeres from the 79 surplus embryos with probes specific for chromosomes 13, 18, 21, X, and Y. QF-PCR was performed with each one or two of the primers specific for the same chromosomes on 151 blastomeres from the same embryos obtained from patients undergoing IVF treatment. RESULTS: Analyses were possible on 135 blastomeres (93%) by FISH and on 117 blastomeres (77%) by QF-PCR. Of 65 embryos, which could be analyzed by both methods, 20 embryos (31%) were diagnosed as abnormal. CONCLUSION: The present study shows that FISH tests are more accurate than QF-PCR assays for the detection of numerical chromosome disorders when performed on single blastomeres.

Aneuploidy↗
PubMed2003

Very early prenatal diagnosis on coelomic cells using quantitative fluorescent polymerase chain reaction.

Quantitative fluorescent PCR (QF-PCR) has been shown to be an accurate assay for the rapid prenatal diagnosis of chromosome disorders. The extra-embryonic coelom develops during week 4 of gestation and it can be aspirated from the following week, making coelocentesis the earliest possible method of prenatal diagnosis after implantation. The possibility of using the QF-PCR assay performed on DNA extracted from cells present in the extra-embryonic coelom has been evaluated for the detection of aneuploidies. QF-PCR amplification using several markers for chromosomes X, Y, 21, 18 and 13 was successfully achieved on all 17 serial samples of exo-coelomic fluid (ECF), placental tissue and maternal blood. Multiplex analyses of maternal blood samples and chorionic tissues allowed the distinction of fetal from maternal patterns and, eventually, the identification of maternal contamination of the ECF samples. Prenatal detection of fetal gender was successful in all cases. When tested with autosomal primers, seven samples were found to contain exclusively fetal DNA. Eight samples contained small amounts of maternal DNA that did not interfere with the QF-PCR analysis. One fetus showed trisomy 13. QF-PCR requires very small volumes of sample compared with cell culture, suggesting that coelocentesis may prove useful for very early prenatal diagnosis.

Aneuploidy↗
PubMed2002

X chromosome dosage by quantitative fluorescent PCR and rapid prenatal diagnosis of sex chromosome aneuploidies.

During the past few years, rapid prenatal diagnosis of chromosome aneuploidies has been successfully achieved by quantitative fluorescent PCR (QF-PCR) amplification of chromosome-specific small tandem repeats (STR). This approach has proven to be very useful in clinical settings, since it allows the detection of major numerical disorders in a few hours after sampling. For the detection of Turner's syndrome (45,X), several highly polymorphic STR on the X chromosome are needed in order to reduce the likelihood that a normal female might be homozygous for all sequences and, consequently, that the test could fail to discriminate between samples retrieved from a Turner's and a normal female fetus. Here we report a new method for rapid and accurate detection of X chromosome copy number in prenatal samples that does not depend on STR heterozygosity. The test is based on QF-PCR amplification of the X-linked HPRT together with the autosomal D21S1411 used as internal control for quantification. In the course of this study, this assay allowed the prenatal diagnosis of a rare case of a normal female homozygous for four selected highly polymorphic X chromosome STR, as well as the assessment of the normal chromosome complement of a fetus homozygous for five chromosome 21 markers.

Amniocentesis↗