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1.
Biochimica Clinica ; 46(3):S184, 2022.
Article in English | EMBASE | ID: covidwho-2168225

ABSTRACT

Background At the Urbino hospital, all patients tested with antigen tests for SARS-CoV-2, if positive, are confirmed with a molecular test. Among the molecular tests in use in our laboratory, VitaPCRTM SARS-CoV-2 Assay is the fastest and easiest to use, as it takes about half an hour from preparation to the result and does not require a biosecurity laboratory, because performed from inactivated samples. Aim of the study Given the rapidity of VitaPCRTM SARS-CoV-2 Assay, it was decided to use it for the confirmation of samples found positive to LIAISON SARS-CoV-2 Ag test. This last test is performed by dedicated tubes with inactivating buffer based on detergent, therefore it does not interfere in the processes of extraction and amplification of nucleic acids. Materials and methods For evaluation, 48 samples tested positive with LIAISON SARS-CoV-2 Ag (Diasorin) test were analyzed, coming from various departments of the Urbino hospital, and confirmed by SimplexaTM COVID-19 Direct (Diasorin) molecular test. From the tubes of all the samples, 200 uL were taken and transferred to dedicated tubes for analysis with VitaPCRTM SARS-CoV-2 Assay (Menarini) to be analyzed, after eliminating 2.2 mL of buffer. Results VitaPCRTM SARS-CoV-2 Assay was able to detect SARSCoV- 2 RNA in all 48 tested antigenically positive samples. In particular, even in all positive samples with low values of TCID50/mL (<1000), 11 out of 48, results with Ct<28.8 were found, although not maintaining a direct correspondence between quantity of antigen and quantity of detected RNA. Conclusions The results obtained, despite requiring an increase in the number of cases, demonstrate the potential use of VitaPCRTM SARS-CoV-2 Assay for the confirmation of positive samples directly from the inactivated buffer used for LIAISON SARS-CoV-2 Ag assay. VitaPCRTM instrument, given the speed, simplicity of use and the possibility of use outside biosafety laboratories, it is an excellent diagnostic tool to support the confirmation procedures of samples tested positive for the SARSCoV- 2 N antigen.

2.
Biochimica Clinica ; 46(3):S118, 2022.
Article in English | EMBASE | ID: covidwho-2168105

ABSTRACT

Since September 2020 the current global pandemic of COVID-19 caused by the SARS-CoV-2 coronavirus is characterized by a succession of waves of infection due to the emergence of new variants of the original virus, presenting various genomic mutations. Many mutations are present in the gene encoding the Spike protein, the main target of the nucleic acidbased vaccines. The Variants of Concern that have been reported since autumn 2020 include Alpha/ B.1.1.7 and sublineages, Beta/B.1.351, Gamma/P.1 and sublineages, Delta/B.1.617.2 and sublineages, Omicron/ B.1.1.529 and sublineages. The rapid and cheap variant monitoring in the population is pivotal for epidemiological studies and for the prompt detection of SARS-CoV-2 variants characterized by high transmissibility or reduced susceptibility to neutralizing antibodies induced by vaccination. Surveillance of genomic variants is currently based on viral whole genome sequencing (WGS) performed on a random fraction of samples positive to molecular tests. WGS involves high costs and extended analysis time compared to a PCR-based diagnostic test, as well as specialized staff and expensive instruments. To rapidly identify the variant in samples positive to SARS-CoV-2, different rapid tests based on real-time PCR and high-resolution melting (HRM) were designed and applied on 88 oropharyngeal swab samples collected from October 2020 to February 2022 (84 positive samples and 4 negative samples). The HRM results were confirmed by PCR product sequencing. Overall, the assays showed 100% specificity and sensitivity compared with commercial PCR assay for COVID-19 testing. Moreover, 83 samples out of 84 (98.8%) were correctly identified as follows: 8 Wuhan (wild type), 12 Alpha, 23 Delta, 37 Omicron BA.1, 1 Omicron BA.1.1, 2 Omicron BA.2. With our lab equipment, about 10 samples can be processed every 3 hours at the cost of 8.5 per sample, including RNA extraction. The identified variants overlapped with mutation and case prevalence over time in Italy (as reported in outbreak.info, which collects genomic data from the GISAID Initiative), accounting for the feasibility of this approach.

3.
Biochimica Clinica ; 45(SUPPL 2):S48, 2022.
Article in English | EMBASE | ID: covidwho-1733403

ABSTRACT

Background The diagnosis of SARS-CoV-2 infection is essential for the control of the epidemic, the establishment of protecting measures and the therapeutic management of patients. Among diagnostic tests utilizing a real-time reverse transcription polymerase chain reaction (RTPCR) amplification technology, VitaPCRTM SARS-CoV-2 Assay was used in our Laboratory for the rapid qualitative detection and discrimination of SARS-CoV-2 viral RNAs in direct nasopharyngeal (NP) or oropharyngeal (OP) swabs. Aim of the study To test SARS-CoV-2 Assay on NP or OP swabs collected in UTM instead of directly melted in the provided buffer of the kit in order to assess the detection efficiency of the system in relation to the potential interference of UTM in the PCR reaction and to viral RNA concentration. Methods A pool of ten SARS-CoV-2 positive samples collected in UTM, previously tested by quantitative real-time RT-PCR, was assayed on the VitaPCRTM Instrument at several dilutions (from 1:10 to 1:100000) in the provided buffer of the kit, immediately and after a cycle of freezing and thawing at -20°C. Results VitaPCRTM Instrument was able to detect SARSCoV- 2 viral RNA at four out of five tested dilutions, with a good efficiency especially at 1:10 and 1:100 dilutions, while at 1:1000000 the result was doubtful. After freezing and thawing the detection efficiency sensibly decreased and the Instrument was able to detect the viral RNA only at 1:10, 1:100 and 1:1000 dilutions, while at 1:10000 and 1:1000000 the result was doubtful. Conclusions These preliminary results are promising suggesting the potential use of SARS-CoV-2 Assay on the VitaPCRTM Instrument even with swabs collected in UTM, increasing the versatility of this system. Moreover, the reduced performance of VitaPCRTM Instrument after freezing and thawing the sample suggests a consistent degradation of viral RNA, advising against the use of this type of frozen samples with this assay. Further investigations are recommended.

4.
Biochimica Clinica ; 45(SUPPL 2):S48, 2022.
Article in English | EMBASE | ID: covidwho-1733036

ABSTRACT

The current global pandemic (COVID-19) caused by the new Betacoronavirus SARS-CoV-2 is characterized by successive waves of infection due to new variants that include mutations in the gene encoding the Spike protein, the main target of the nucleic acidbased vaccines. In fact, as of autumn 2020, several countries have reported the detection of SARS-CoV-2 variants that have spread more efficiently (referred to as variants of concern by WHO). Such variants include the Alpha variant (English variant, B.1.1.7), the Beta variant (South African variant, B.1.351), the Gamma variant (Brazilian variant, P.1), and the more recent Delta variant (Indian variant, B.1.617. 2). Therefore, it is pivotal to monitor the virus and the onset of SARSCoV-2 variants characterized by high transmissibility or reduced susceptibility to neutralizing antibodies induced by vaccination.Surveillance of genomic variants is currently based on sequencing of viral genomes performed on a random fraction of samples positive by molecular test. The sequencing of 228 SARS-CoV-2 positive samples by ASUR Marche Area Vasta 1 (Fano-Pesaro-Urbino) from February to June 2021 highlighted the progressive increase of variants (mainly B.1.1.7 and to a lesser extent P.1) from early February until March 18th. From March 18th onwards, only variants B.1.1.7 and P.1 were detected. DNA sequencing involves high costs and extended analysis time compared to a PCR-based diagnostic test. To rapidly identify the samples containing virus variants to be sequenced for complete characterization, in synergy with the University of Urbino, five rapid tests based on real-time PCR and high-resolution melting (HRM) were designed on the gene encoding the Spike protein. Preliminary results indicated that the sensitivity of the assays was not significantly different from that of commercial molecular tests. Furthermore, through HRM analysis, it was possible to discriminate amplicons with mutation 1709 C > A causing the amino acid substitution A570D, specific for the alpha variant.

6.
European Heart Journal, Supplement ; 23(SUPPL C):C81, 2021.
Article in English | EMBASE | ID: covidwho-1408957

ABSTRACT

Background: MoviS: 'Movement and health beyond care' is an ongoing randomized controlled trial aiming to educate breast cancer (BC) survivors on the benefits of exercise and proper nutrition habits. Methods: The study included thirty women (17.4% of the total planned cohort of 172 patients) with stage 0-III non-metastatic BC (age: 53.5±7.6 yrs;BMI: 25.3±4.9 kg/ m2) randomly allocated to the Intervention Arm (IA;supervised exercise training: “MoviS Training”) or control arm. The MoviS Training consists of 12 weeks of aerobic exercise (2 d/week of supervised and 1 d/week of unsupervised exercise) which intensity and duration gradually increased from 40 to 70% of heart rate reserve and from 20 to 60min, respectively. Both arms received nutritional and lifestyle counselling based on WCRF 2018 guidelines through the DIANA-Web platform and motivational interviewing. As the planned protocol was changed due to nationwide lockdown to contain the spread of COVID-19, IA performed home-based exercise sessions, which were remotely supervised using heart rate monitors. Heart rate variability (HRV;by 24-Holter monitoring), cardiac function indexes (by echocardiography with speckle tracking imaging), and cardiorespiratory fitness (by estimated maximal oxygen uptake [VO2max]) were evaluated at baseline and after the intervention period. Results: There were no adverse events during training. Baseline evaluation revealed no systolic disfunction (mean LVEF 60.4±4.5%) and a mild reduction (values 3 -18%) in global longitudinal strain in 26% of patients. HRV improved in both time and frequency domains: ASDNN/5min (50.6±14.4 to 55.2±16.7 msec, p=0.033);very low frequency (VLF) (1597±967 to 1881±963 msec, p=0.04);low frequency (LF) (613±404 to 731±542 msec, p=0.004);total power (2627±1393 to 3034±1669 msec, p=0.034). HRV parameters tended to improve to a greater extent in IA group (Coefficient of Variation: ASDNN/5min 13.7% vs 4.6%, LF 27% vs 10%, total power 26.5% vs 5.1%). Cardiorespiratory fitness level increased significantly in both groups (VO2max from 30.7±5.7 to 33.9±6.6mL/kg/min, p<0.001). Conclusion: During COVID-19 lockdown, short-term remotely supervised exercise training and recommendations on a healthy lifestyle lead to a significant improvement in HRV parameters and cardiorespiratory fitness in BC survivors.

7.
Tumori ; 106(2 SUPPL):212-213, 2020.
Article in English | EMBASE | ID: covidwho-1109821

ABSTRACT

Background: Breast cancer (BC) is the most common invasive cancer in women and evidence has shown that exercise can significantly improve the outcomes of BC survivors. MoviS: 'Movement and Health Beyond Care' is a randomized controlled trial, which aims to educate cancer patients after adjuvant therapy on the benefits of exercise and proper nutritional plan. Material (patients) and methods: Thirty women with stage 0-III non-metastatic BC recruited 12-month postsurgery (age: 53.5±7.6;BMI: 25.3±4.9) were randomized in two groups: intervention arm received 3-month aerobic training (40-70% of the heart rate reserve;20 to 60 min 3-day/week);control arm with usual care recommendations. Patients enrolled in January 2020 were monitored for diet habits by DianaWeb platform, they carried out the exercise training across the COVID-19 pandemic shifting from the gym to a home-based exercise program. The objective was to investigate the effects of lifestyle program on quality of life by EORTC QLQ-C30 and health-related QoL parameters such as cardiac function indexes;heart rate variability (HRV) (by repeated Holter 24h) and cardiorespiratory fitness by estimated maximal oxygen uptake (VO2max). Results: There were no adverse events and a high attendance was registered considering the challenges to be overcome in shifting to a home-based exercise program. Statistical analysis revealed a significant improvement in both arms in QLQ scale score: in global health status (from 64.7±17 to 15.9±13;coefficient of variation (CV) 15.9%;p=0.0015);physical functioning (from 54.4±12.3 to 6±6.6;CV 13.9%;p=0.0005);fatigue (from 26.3±23.4 to 11.9±14.3;CV -54.9%;p=0.0008) and showed a general improvement over time even on the social functioning (from 47.2 ± 22.8 to 66.7±00;CV 41.2%;p=0.0001). Both groups had a significant improvement in HRV parameters in both time and frequency domain: average SDNN/5min and VLF increased (from 50.6±14.4 to 55.2±16.7msec, p=0.033 and form 1597±967 to 1881±963 msec, p=0.04, respectively);mean heart rate decreased (from 76.6 ±7.8 to 73.7±8.3 bpm after training, p=0.009) and both arms improved the cardiorespiratory fitness level (VO2max from 30.7±5.7 to 33.9±6.64 mL/kg/min;CV 10.3%;p<0.001). Conclusions: Despite the challenges faced during the pandemic, a change in lifestyle integrating exercises, nutrition and educational counseling provides benefit to BC patients when delivered in a clinic-based setting group including the exercise specialists.

9.
Annals of Oncology ; 31:S335-S336, 2020.
Article in English | EMBASE | ID: covidwho-801214

ABSTRACT

Background: MoviS: ‘Movement and Health Beyond Care’ is a 3-year randomized controlled trial aiming to educate cancer patients after adjuvant therapy on the benefits of personalized physical activity (PA) and a proper nutritional plan. Methods: In this project, breast cancer (BC) patients who consent to participate will be randomized to Interventional Arm (A), consisting of 3-months of Movis Training, or Control Arm (B), consisting of standard care with no supervised PA. The Movis Training consists of 3-months of aerobic training (2 d/week of supervised training and 1 d/week of unsupervised exercise) with an increase in exercise intensity (40-70% Heart Rate Reserve) and duration (20-60 min). Ad interim analyses every 3-months up to 1-year will be included. The first cohort of eligible BC survivors were enrolled in January 2020 and carried out the Movis Training even during the COVID-19 pandemic. The primary outcome is improvement of Quality of Life (QoL) assessed by European Organization for Research and Treatment of Cancer QoL (EORTC QLQ-C30). The secondary outcomes are improvement of health-related QoL parameters such as: PA level (International Physical Activity Questionnaire;SenseWear Armband), fitness (VO2max), flexibility, strength, psychological well-being (Psychological distress Inventory;Profile of Mood States and diet habit (DianaWeb, MEDIET modified and recall 24h);Anthropometric measurement, Body mass (kg);BMI (kg/m2);body composition. Results: The expected improvement (mean ± SD) of the QoL in Arm A at 3-months is 15.1 ± 17.7, while in Arm B is 6.1 ± 17.1 (Cohen d effect size=0.51, medium effect). Using a t test for independent samples, with 0.05 alpha and 0.80 1-beta will require 60 subjects per group. Considering an expected drop-out of 30%, a total of 172 patients will be recruited. Conclusions: The targeted exercise oncology through multidisciplinary team would like to provide a coordinated program of cancer care to improve health care quality, improve prognosis, increase survival times and QoL and reduce the risk of BC recurrence. Legal entity responsible for the study: Elena Barbieri. Funding: Has not received any funding. Disclosure: All authors have declared no conflicts of interest.

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