Evaluation of extraction-free RT-PCR methods for faster and cheaper detection of SARS-CoV-2 using two commercial systems.

Objective: When using high-throughput batched diagnostic platforms based on RT-PCR for SARS-CoV-2 detection, avoidance of the conventional nucleic acid extraction step can help to reduce the turnaround time and increase processivity. This approach can also spare reagents and plasticware, which have experienced a shortage during the initial waves of the pandemic, reducing the overall testing costs.

Methods: This study evaluated the performance of extraction-free protocols based on simple dilution of the specimen in sterile RNAse free water (with or without a heating step) in comparison to standard RNA extraction protocols, using two commercial kits for molecular detection of SARS-CoV-2 (Allplex™ SARS-CoV-2 assay and Allplex™ SARS-CoV-2/FluA/FluB/RSV assay) in nasopharyngeal swabs (NPS).

Hand, foot, and mouth disease in pregnancy: 7 years Tuscan experience and literature review.

Objective: Evaluation of hand, foot, and mouth disease (HFMD) diagnostic strategies in pregnancy and the risk of HFMD-related fetopathy.

Study Design: Pregnant women consecutively evaluated between 2010 and 2016 at the Tuscany Reference Center for Infectious Diseases in Pregnancy for HFMD were enrolled. A descriptive analysis of infected patients/newborns data and literature review were carried out.

Evaluation of a new rapid fluorescence immunoassay for the diagnosis of dengue and Zika virus infection.

Background: Dengue (DENV) and Zika virus (ZIKV) are important mosquito-transmitted viruses.

Objectives: To investigate the performance of Standard F, Fluorescence Immunoassay (FIA, SD Biosensor Inc., Suwon, South Korea) providing results in 15 min to detect DENV IgG, IgM and NS1Ag, and ZIKV IgG, IgM, and Ag.

Cryptic severe malaria in a Moroccan man living in Tuscany, Italy, August 2018.

In August 2018 a Moroccan man living in Tuscany developed malaria. The patient declared having not recently visited any endemic country, leading to diagnostic delay and severe malaria. As susceptibility to of Anopheles species in Tuscany is very low, and other risk factors for acquiring malaria could not be completely excluded, the case remains cryptic, similar to other malaria cases previously reported in African individuals living in Apulia in 2017.

Regional spread of contact lens-related Acanthamoeba keratitis in Italy.

Acanthamoeba ocular infections, known as Acanthamoeba keratitis, are an emerging problem among contact lens wearers. Infections mediated by Acanthamoeba are uncommon, but they can be underestimated due to poor awareness and delayed diagnosis. The routine use of rapid and cost-effective molecular methods like Real Time PCR for the diagnosis of this important pathogen could improve diagnosis and therapy outcome.

Utility of droplet digital PCR for the quantitative detection of polyomavirus JC in clinical samples.

Background: Quantitative PCR (qPCR) is the standard molecular method for detection of polyomavirus JC (JCPyV) DNA reactivation in serum and cerebrospinal fluid (CSF) in patients at risk of progressive multifocal leukoencephalopathy (PML). Recently, digital PCR has shown potential benefits over qPCR in viral diagnostics.

Objective: To evaluate the performance of droplet digital PCR (ddPCR) assay in assessing JCPyV-DNA status in clinical samples of patients at risk for PML.

Detection of 12 respiratory viruses by duplex real time PCR assays in respiratory samples.

Different viruses can be responsible for similar clinical manifestations of respiratory infections. Thus, the etiological diagnosis of respiratory viral diseases requires the detection of a large number of viruses. In this study, 6 duplex real-time PCR assays, using EvaGreen intercalating dye, were developed to detect 12 major viruses responsible for respiratory diseases: influenza A and B viruses, enteroviruses (including enterovirus spp, and rhinovirus spp), respiratory syncytial virus, human metapneumovirus, coronaviruses group I (of which CoV 229E and CoV NL63 are part) and II (including CoV OC43 and CoV HKU1), parainfluenza viruses type 1, 2, 3 and 4, human adenoviruses and human bocaviruses.