Rapid monitoring of SARS-CoV-2 variants through high resolution melt analysis

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.

Severe Neutropenia after Sarilumab Administration in Two COVID-19 Patients: Case Reports and Literature Review

BACKGROUND: Two years have passed since the WHO declared a pandemic state for SARS-CoV2 infection. COVID-19 pathogenesis consists of a first viral phase responsible for early symptoms followed by an inflammatory phase, which is cytokine-mediated, responsible for late-onset signs up to acute respiratory distress syndrome. Considering that interleukin (IL)6 plays a key role in the development and maintenance of inflammation, drugs targeting both IL6 and IL6 receptors have been evaluated. CASE REPORTS: The present study reports the cases of two hospitalized patients with severe respiratory COVID-19 treated with a single dose of intravenous sarilumab, a monoclonal anti-IL6 antibody, along with standard of care medications and high-flow oxygen therapy. Although a few days following sarilumab administration, clinical and biochemical conditions started ameliorating, these patients developed severe and self-limiting neutropenia. CONCLUSION: Sarilumab may represent a promising weapon to treat the fearsome hyperinflammatory phase;however, more trials are needed to decide whether to use it in combination with other drugs or alone, and to better understand pharmacokinetics and side effects.

Pain crisis management in a patient with sickle cell disease during SARS-CoV-2 infection: A case report and literature review

Patients with sickle cell disease (SCD) are more susceptible to severe coronavirus disease 2019 (COVID-19) infection, in comparison with the general population, due to the possibility that the inflammatory state, along with hypoxia and hypercoagulability may increase the risk of developing acute SCD-related complications. The present study reports the case of a 33-year-old female affected by SCD, who although vaccinated against COVID-19, tested positive for SARS-CoV-2 and developed febrile pneumonia. During hospitalization, the patient complained about generalized intense pain, along with fever recurrence and increased inflammatory marker, procalcitonin and haemoglobin S levels. The patient was treated with an intravenous analgesic therapeutics cocktail in combination with red blood cell manual exchange procedure and broad-spectrum antibiotic therapy, achieving the rapid resolution of pain and an improvement in the laboratory test results. From the case presented herein, it is thus suggested that patients with SCD and COVID-19 infection need to be critically evaluated by clinicians, as such patients may develop severe outcomes, attributed to the overlap of two difficult to treat conditions. © 2022 by the authors.

Development of qPCR tests for monitoring SARSCoV-2 variants through high resolution melting analysis

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.

A robotized hospital bed for COVID-19 patients in intensive care treatments

Patients in bed for clinical situations and specially in COVI19 treatments need special assistance with suitable bed configuration motion both to help therapies and physical conditions. Hospital beds are designed to ensure static safe configuration of patients mostly in supine position with some motion capability in elevating trunk or legs. More capability is today requested for permitting bedded patients to avoid inconveniences up to further accidents and to facilitate therapy treatments and clinical conditions. A robotized design solution for hospital beds with movable segments is proposed for patients of COVID-19 in intensive and sub-intensive care treatments with the aim to help therapies and motion treatments for better physical conditions. The proposed design is analyzed in terms of mechanism structure and motion operation towards medical effects that can be appreciated as supporting/mitigating critical conditions in hospital applications. © 2021 IEEE.

The role of the SARS-COV 2 pandemic on the delay of diagnosis in a case of multiple myeloma associated with AL amyloidosis in HIV-HBV positive patient on antiretroviral treatment

COVID-19 has devastated healthcare systems all over the world and is still raging in Italy. In many countries, patients with chronic illnesses are suffering from delay in diagnostic and treatment management. We report a challenging case of a HIV patient who experienced delay in diagnosis of multiple myeloma due to COVID-19 pandemic restrictions. Global cooperation to ensure equity and responsiveness to local contexts is essential on the difficult path ahead to ending the COVID-19 pandemic, as treatment for one potentially curable disease should not be performed at the expense of another.

High-flow nasal cannula oxygenation and tocilizumab administration in patients critically ill with COVID-19: A report of three cases and a literature review

Since late 2019, SARS-CoV2 has spread worldwide, leading the WHO to declare a pandemic state. Italy was deeply affected by the virus, particularly North Italy. Several molecules have been tested for the treatment of coronavirus disease (COVID-19), comparing the treatment efficacy and collateral effects. To date, no antiviral drugs have been approved for the treatment of the COVID-19 viral phase or for the inflammatory phase. Undoubtedly, oxygen support plays a key role in the management of patients affected by this virus. The present study reports the cases of 3 patients critically ill with COVID-19. Despite antiviral therapy, their clinical conditions deteriorated a few days following admis- sion, particularly as regards respiratory performance, together with chest X-ray findings and arterial blood gas parameters. The levels of inflammatory markers were also elevated. The patients were treated with high-flow nasal cannula (HFNC) oxygenation along with a double dose of tocilizumab. A few days following HFNC and tocilizumab administration, the respiratory rates and arterial blood gas data were ameliorated alongwithchestX-rayresults.TheuseofHFNCwasthenslowly reduced until it was terminated, with the patients achieving a successful discharge. On the whole, as presented herein, it is indisputable more data and guidelines for COVID-19 therapies are warranted in order to guide clinicians as to the appropriate clinical treatment which will guarantee an optimal therapeutic response. © 2020 World Academy of Sciences Journal. All rights reserved.

Driving Mechanism in Robotized Hospital Bed for Patients with COVID 19

This paper presents a mechanism solution for a robotized bed with movable segments also for lateral movements of patients, the feasibility of the proposed mechanism solution is discussed by using simulation results for a typical operation. The main focus is addressed on mechanical aspects that nevertheless are affected by control actions and sensor feedbacks. © 2021, The Author(s), under exclusive license to Springer Nature Switzerland AG.

Containment effort reduction and regrowth patterns of the Covid-19 spreading.

In all countries the political decisions aim to achieve an almost stable configuration with a small number of new infected individuals per day due to Covid-19. When such a condition is reached, the containment effort is usually reduced in favor of a gradual reopening of the social life and of the various economical sectors. However, in this new phase, the infection spread restarts and, moreover, possible mutations of the virus give rise to a large specific growth rate of the infected people. Therefore, a quantitative analysis of the regrowth pattern is very useful. We discuss a macroscopic approach which, on the basis of the collected data in the first lockdown, after few days from the beginning of the new phase, outlines different scenarios of the Covid-19 diffusion for longer time. The purpose of this paper is a demonstration-of-concept: one takes simple growth models, considers the available data and shows how the future trend of the spread can be obtained. The method applies a time dependent carrying capacity, analogously to many macroscopic growth laws in biology, economics and population dynamics. The illustrative cases of France, Italy and United Kingdom are analyzed.

Transmission mode associated with coronavirus disease 2019: a review.

OBJECTIVE: In late December 2019 in Wuhan (China), Health Commission reported a cluster of pneumonia cases of unknown etiology, subsequently isolated and named Severe Acute Respiratory Syndrome (SARS) Coronavirus 2 (CoV-2). In this review, the main transmission routes and causes of mortality associated with COVID-19 were investigated. MATERIAL AND METHODS: A review was carried out to recognize relevant research available until 10 April 2020. RESULTS: The main transmission routes of COVID-19 have been the following: animal to human and human-to-human pathways, namely: respiratory transmission; oro-fecal transmission; air, surface-human transmission. Transmission from asymptomatic persons, healthcare transmission, and interfamily transmission have been well documented. CONCLUSIONS: SARS-CoV-2 possesses powerful pathogenicity and transmissibility. It is presumed to spread primarily via respiratory droplets and close contact. The most probable transmission pathway is definitely the inter-human one. Asymptomatic patients seem to play a crucial role in spreading the infection. Because of COVID-19 infection pandemic potential, careful surveillance is essential to monitor its future host adaptation, viral evolution, infectivity, transmissibility, and pathogenicity in order to gain an effective vaccine and flock immunity and reduce mortality as soon and as much as it is possible.

Outbreak of COVID-19 infection in children: fear and serenity.

OBJECTIVE: The recent outbreak of SARS-CoV-2 greatly involves the resources of the global healthcare system, as it affects newborns, adults, and elders. This infection runs in three major stages: a mild cold-like illness, a moderate respiratory syndrome and a severe acute interstitial pneumonia. SARS-CoV-2 infection seems to have a more benign evolution in children. As a matter of fact, low susceptibility and minor aggressivity have been highlighted in most cases. There are currently no effective antiviral drugs treatment for the affected children. No sufficient results have been reached by the use of interferon (IFN), lopinavir/ritonavir, orbidol, and oseltamivir in the treatment of the coronaviruses infection. The aim of this short review is to highlight the differences existing between COVID-19 cases in adults and children.

Eculizumab treatment in patients with COVID-19: preliminary results from real life ASL Napoli 2 Nord experience.

OBJECTIVE: SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2)-related pneumonia, referred to as COVID-19 (Coronavirus Disease 19), is a public health emergency as it carries high morbidity, mortality, and has no approved specific pharmacological treatments. In this case series, we aimed to report preliminary data obtained with anti-complement C5 therapy with eculizumab in COVID-19 patients admitted to intensive care unit (ICU) of ASL Napoli 2 Nord. PATIENTS AND METHODS: This is a case series of patients with a confirmed diagnosis of SARS-CoV2 infection and severe pneumonia or ARDS who were treated with up to 4 infusions of eculizumab as an off-label agent. Patients were also treated with anticoagulant therapy with Enoxaparin 4000 IU/day via subcutaneous injection, antiviral therapy with Lopinavir 800 mg/day + Ritonavir 200 mg/day, hydroxychloroquine 400 mg/day, ceftriaxone 2 g/day IV, vitamine C 6 g/day for 4 days, and were on Non-Invasive Ventilation (NIV). RESULTS: We treated four COVID-19 patients admitted to the intensive care unit because of severe pneumonia or ARDS. All patients successfully recovered after treatment with eculizumab. Eculizumab induced a drop in inflammatory markers. Mean C Reactive Protein levels dropped from 14.6 mg/dl to 3.5 mg/dl and the mean duration of the disease was 12.8 days. CONCLUSIONS: Eculizumab has the potential to be a key player in treatment of severe cases of COVID-19. Our results support eculizumab use as an off-label treatment of COVID-19, pending confirmation from the ongoing SOLID-C19 trial.