Deletions on the short arm of the Y chromosome involving the amelogenin Y gene (AMELY), located on Yp11.2, can be misleading for sex typing with serious consequences in forensic applications and prenatal diagnosis. In this study, we describe two AMELY null cases concerning two unrelated Italian males from Northeast Italy. PCR amplification of short tandem repeats on the Y chromosome (Y-STRs) showed a lack of AMELY and DYS458 markers. The presence of all the other markers located on the Y chromosome and of the SRY gene in both samples led us to conclude that a deletion had occurred in a portion of the short arm of the Y chromosome. Twenty-three Y-specific sequence tagged sites (STSs) were chosen to delineate the deletion's length, which was estimated to be in the range of 3.35–3.87 Mb for one sample and 1.51–2.58 Mb for the other. These and previous findings suggest that in all cases where potential AMELY drop out has occurred, it should be used additional specific Y chromosome markers or human DNA quantification methods that specifically quantify male DNA using target male genomic markers, which not being located within the deletion regions, allow an accurate sex identification.

Despite being deliberately targeted to common chromosome aneuploidies, the rapid quantitative fluorescent polymerase chain reaction (QF‐PCR) tests can detect the majority of chromosome abnormalities in prenatal diagnosis. The main advantages of this assay are low cost, speed and automation allowing large‐scale application.

OBJECTIVE:We surveyed the datasheets of 29 laboratories concerning prenatal diagnosis of de novo apparently balanced chromosome rearrangements to assess the involvement of specific chromosomes, the breakpoints distribution and the impact on the pregnancy outcome. METHOD:By means of a questionnaire, data on 269.371 analyses performed from 1983 to 2006 on amniotic fluid, chorionic villus and fetal blood samples were collected. RESULTS:A total of 246 balanced anomalies were detected at frequencies of 72% for reciprocal translocations, 18% for Robertsonian translocations, 7% for inversions and 3% for complex chromosome rearrangements. The total frequencies of balanced rearrangements were 0.09%, 0.08% and 0.05% on amniotic fluid, chorionic villus and fetal blood samples. CONCLUSION:A preferential involvement of chromosomes 22, 7, 21, 3, 9 and 11 and a less involvement of chromosomes X, 19, 12, 6 and 1 was observed. A nonrandom distribution of the breakpoints across chromosomes was noticed. Association in the location of recurrent breakpoints and fragile sites was observed for chromosomes 11, 7, 10 and 22, while it was not recorded for chromosome 3. The rate of pregnancy termination was about 20%, with frequencies decreasing from complex chromosomal rearrangements (33%), reciprocal translocations (24%) to inversions (11%) and Robertsonian translocations (3%).

Background: Human Papillomavirus (HPV) infection has been well established as etiological agent of cervical cancers. Introduction of HPV genotyping either in cervical screening programs or for monitoring the effectiveness of HPV vaccination, requires access to fast, simple and high-throughput technology.Objectives: Our objective was to evaluate a new multiplex fluorescent PCR method for HPV detection and typing assessing high risk HPV frequency and relationship with cytological and histological data.Materials and Methods: Cervical specimens were collected from 4,051 Italian women (15-65 years aged ) with abnormal cytological smears and were tested for the presence of HPV using the f-HPV typing Kit, a recently available commercial assay (Molgentix Barcelona). Extracted DNA was amplified with 15 sets of fluorescent labelled primers recognising HPV type 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 68, 6 and 11. Labelling of primers with 6-FAM, NED, VIC and PET allowed high sensitivity as well as amplification of similar size products in the same PCR reaction. After electrophoresis on ABI 3130 DNA sequencer, PCR products were automatically recognised by size and colours. Cytological and histological analysis was performed.Results: High risk HPV types were detected in 80% (1,145/4,091) and low risk HPV in 20% (284/4,091) of the samples. High risk HPV were detected in 42% (43/102) of ASCUS-AGUS, 50% (234/466) of CIN1, 70% (88/126) of CIN2 , 97% (35/36) CIN3, 98% (118/120) of squamous cervical cancer and finally in 100% (25/25) of adenocarcinomas. HPV-16 was the most prevalent type in LSILs (40%), in HSILs (72%), and in carcinomas (62%) followed by HPV-31.Conclusions: The f-HPV typing™ kit is an accurate technique for quick and sensitive detection of 15 HPV types, it also allows distinguishing single and multiple infections in several cases. The high throughput of the assay allows assessing different HPV types frequency in a population and may be of help in determining possible changes that could derive as consequence of vaccination policies.

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.

Sex tests based on amelogenin are part of various PCR multiplex reaction kits widely used for human gender identification and have important applications in forensic casework, prenatal diagnosis, DNA databasing and blood sample storage. The two most common sex tests based on amelogenin are represented by primer sets that delimit a 6-bp deletion on the X chromosome to produce X/Y fragments of 106/112 or 212/218 bp, respectively. Few cases of AMELY deletion, usually considered as polymorphisms, have been reported so far and a detailed characterization of the molecular alteration is still lacking. In this study, we describe a large interstitial deletion of the Y short arm encompassing the AMELY locus in two unrelated individuals. The first case was identified in an oligozoospermic, otherwise phenotypically normal, 32-year-old man during the screening for Y microdeletions performed on a sample of infertile males. The second one was found among amniotic liquid samples tested by quantitative fluorescence-polymerase chain reaction and cytogenetic analysis for prenatal diagnosis. The extent of the deletion, spanning approximately 2.5 Mb, was better characterised by pulsed-field gel electrophoresis, followed by fluorescence in situ hybridization and STS marker analysis.

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 rearrangements can lead to aneuploidies of specific chromosome regions and could be present in the entire individual or limited to some tissues (mosaicism) depending on the developmental stage of the embryo when the rearrangement occurs. We report 6 cases with sex chromosome rearrangements identified by conventional cytogenetics and tested by quantitative fluorescent polymerase chain reaction (QF-PCR). QF-PCR has been largely employed for rapid detection of common aneuploidies in pre-natal and post-natal diagnosis and consists in DNA amplification by polymerase chain reaction (PCR) using fluorescent labelled primers and the analysis of chromosome specific small tandem repeats (STR). We tested 5 sex chromosome specific STR markers in multiplex PCR amplifications together with other chromosome specific STR markers as control amplifications. The PCR products were analysed by capillary electrophoresis. The results from QF-PCR analysis were obtained within one day and confirmed our cytogenetic observations. This study shows that QF-PCR analysis can detect sex chromosome imbalance and also suspect mosaicism or chromosome rearrangement.

Risk factors other than human papillomavirus (HPV) infection per se for cervical cancer development have been investigated recently. It was suggested that HPV 16 E6 variants and the p53 codon 72 arginine polymorphism could be progression markers. Indeed, it has been demonstrated that specific E6 variants and p53 arginine were both enriched in cancer. However, especially with regard to the latter, divergent results have been reported. Our aim was thus to investigate whether p53 arginine is important for cervical carcinogenesis by scaling up samples of the two European cohorts, the initial results of which were reported previously. In addition, we have assessed the occurrence of p53 codon 72 arginine, in combination with specific HPV 16 E6 genotypes. We found p53 arginine to be increased in cancer of both cohorts, consistent with our previous concept. Although specificE6 genotypes increased gradually with the severity of the lesion, p53 arginine was enriched in cancer only. Moreover, the frequency of the arginine allele was similar in groups with differentE6 genotypes. It is concluded that p53 arginine is a risk factor for cervical cancer but probably acts independently of E6 variants.

Human MutationVolume 13, Issue 4 p. 340-340 Mutation and Polymorphism Report A novel BRCA1 mutation (1099 delCA stop codon 328) linked to breast and/or ovarian cancer † Gianfranco Voglino, Gianfranco Voglino Department of Clinical Pathology, Service of Histopathology and Diagnostic Cytology, Molecular Pathology Laboratory, Ospedale Sant'Anna, Turin, ItalySearch for more papers by this authorCinzia Pera, Cinzia Pera Advanced Biotechnology Center-IST, Laboratory of Immunogenetics, Genoa, ItalySearch for more papers by this authorGianbattista Ferrara, Gianbattista Ferrara Advanced Biotechnology Center-IST, Laboratory of Immunogenetics, Genoa, ItalySearch for more papers by this authorLorenzo Silengo, Lorenzo Silengo Department of Genetics, Biology and Medical Chemistry, University of Turin, ItalySearch for more papers by this authorGuglielmo Stefanuto, Guglielmo Stefanuto Department of Genetics, Biology and Medical Chemistry, University of Turin, ItalySearch for more papers by this authorAntonella Marongiu, Antonella Marongiu Department of Clinical Pathology, Service of Histopathology and Diagnostic Cytology, Molecular Pathology Laboratory, Ospedale Sant'Anna, Turin, ItalySearch for more papers by this authorLuciano Fessia, Luciano Fessia Department of Clinical Pathology, Service of Histopathology and Diagnostic Cytology, Molecular Pathology Laboratory, Ospedale Sant'Anna, Turin, ItalySearch for more papers by this author Gianfranco Voglino, Gianfranco Voglino Department of Clinical Pathology, Service of Histopathology and Diagnostic Cytology, Molecular Pathology Laboratory, Ospedale Sant'Anna, Turin, ItalySearch for more papers by this authorCinzia Pera, Cinzia Pera Advanced Biotechnology Center-IST, Laboratory of Immunogenetics, Genoa, ItalySearch for more papers by this authorGianbattista Ferrara, Gianbattista Ferrara Advanced Biotechnology Center-IST, Laboratory of Immunogenetics, Genoa, ItalySearch for more papers by this authorLorenzo Silengo, Lorenzo Silengo Department of Genetics, Biology and Medical Chemistry, University of Turin, ItalySearch for more papers by this authorGuglielmo Stefanuto, Guglielmo Stefanuto Department of Genetics, Biology and Medical Chemistry, University of Turin, ItalySearch for more papers by this authorAntonella Marongiu, Antonella Marongiu Department of Clinical Pathology, Service of Histopathology and Diagnostic Cytology, Molecular Pathology Laboratory, Ospedale Sant'Anna, Turin, ItalySearch for more papers by this authorLuciano Fessia, Luciano Fessia Department of Clinical Pathology, Service of Histopathology and Diagnostic Cytology, Molecular Pathology Laboratory, Ospedale Sant'Anna, Turin, ItalySearch for more papers by this author First published: 31 March 1999 https://doi.org/10.1002/(SICI)1098-1004(1999)13:4<340::AID-HUMU20>3.0.CO;2-3 † Communicated by: Mark H. Paalman ‡ Online Citation: Human Mutation, Mutation and Polymorphism Report #44 (1999) Online http://journals.wiley.com/1059-7794/pdf/mutation/mpr44.pdf § Acknowledgments: We thank the family members who contributed to the study. We acknowledge Claretta Marchi, Domenico Matarozzo and Maria Gugliotta for technical assistance AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Volume13, Issue41999Pages 340-340 RelatedInformation

Estrogen receptors (ER) were evaluated in 634 breast cancer patients by the dextran-coated charcoal method (DCC). In 206, ER and progesterone receptors (PR) were also tested by cytochemistry (Lee method), and in 124 ER were tested by immunofluorescence (Pertschuk method). The median follow-up is 3.7 years.