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1.
DEVELOPMENT OF A NOVEL PCR-BASED ASSAY FOR HIGH-RISK HUMAN PAPILLOMAVIRUS DETECTION AND GENOTYPING IN SELF COLLECTED CERVICOVAGINAL SAMPLES: A NEW POSSIBILITY FOR THE CERVICAL CANCER SCREENING
Alice Avian, 2020, doctoral dissertation

Abstract: Human Papillomavirus (HPV) infection is the causative agent for the invasive cervical cancer and its precancerous lesions, furthermore, there are growing evidences of HPV being a relevant factor in other anogenital cancers as well as head and neck cancers. Most sexually active women become infected with HPV at least once in their lifetime, but less than 10% of women becomes persistently infected, and it is precisely the persistent infection that contributes to the development of cervical cancer. The preventive effect of cervical cancer screening largely depends in the high women participation and coverage; indeed, a large number of cervical cancers diagnoses normally arise among under-screened and unscreened women. Increase in the screening coverage is essential to improve the effectiveness of cervical screening programmes. The main purpose of this PhD project was to solve some of the most relevant problems in the cervical cancer screening programmes, as the increase of cost-effectiveness and the amelioration of the screening coverage. My work was focused on the development and validation of the first Ulisse BioMed S.p.A. product, the HPV Selfy™ test, an innovative PCR-based kit for the direct detection and genotyping of 12 high-risk HPV types (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59) and 2 possible/probable high-risk (66 and 68), specifically optimized for the analysis of self-collected vaginal specimens. The core of this innovative test is based on high-resolution melting (HRM) analysis, a recently developed technique for fast, high-throughput post-PCR analysis of variance in nucleic acid sequences, that characterizes the amplicons by studying thermal denaturation of double-stranded DNA. Based on this approach and through the design of different HPV type-specific primer pairs and the development of a specific master mix, unique melting peaks in a single fluorescence channel were obtained, allowing the multiple detection and genotyping of 14 HPV types in a single PCR well. Three different clinical studies have been carried out to validate the assay on the vaginal self-collected samples with truly amazing results regarding the assay’s performance, but also for self-sampling acceptability by women. Moreover, data collected in these studies suggest a future possible use of this test for the hard-to-reach women, as an alternative of the conventional clinician-collected sample, in order to increase the cervical cancer screening coverage.
Keywords: Human Papillomavirus, HPV test, cervical cancer screening, prevention, diagnostic test, High resolution melting, HRM, genotyping, PCR, Self-sampling, clinical validation.
Published in RUNG: 17.06.2020; Views: 4610; Downloads: 112
.pdf Full text (37,88 MB)

2.
PREVENTION AND MANAGEMENT OF LEGIONELLA SPP. SPREAD IN HOSPITAL WATER SYSTEM (ESTABLISHING AN EFFECTIVE SYSTEM WITHOUT USING CHEMICALS IN UNIVERSITY CLINIC OF RESPIRATORY AND ALLERGIC DISEASES GOLNIK)
Mojca Novak, 2016, master's thesis

Abstract: The hospital water system can be a causative agent for acquiring hospital-acquired infection. Many microorganisms can be detected in potable water and one of this is Legionella spp. These bacteria live also in nature, but are usually present in low numbers. When entering into the buildings and their water supply, Legionella finds ideal conditions to multiply, especially if this systems are extensive and old. To control the conditions in the levels that cannot harm humans, many procedures – physical and chemical – have been established. All of them have advantages as well as limitations. According to these facts, in University Clinic of Respiratory and Allergic Diseases Golnik we tried to reestablish the protocols that would keep us on the safe side and be environmentally and health friendly as well. We have been trying to use the protocol without added chemicals – just with maintaining proper temperatures, heat disinfection and flushing for many years now. With this study we wanted to extend our knowledge of our hospital water system, so we intensified the sampling and introduced extended measures. We found out that the presence of Legionella in water system is connected to moderate temperatures and it is present more frequently in high risk zones (inappropriate temperatures, dead legs, water stagnation). With constant measures, the Legionella can be controlled but not eradicated. To contain it on a safe level, the group of experts must work together and introduce procedures that will provide safe environment for the patients and employees.
Keywords: Legionella spp., hospital water system, prevention, temperature, flushing.
Published in RUNG: 30.09.2016; Views: 7152; Downloads: 288
.pdf Full text (2,81 MB)

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