Buildout and integration of an automated high-throughput CLIA laboratory for SARS-CoV-2 testing on a large urban campus

In 2019, the first cases of SARS-CoV-2 were detected in Wuhan, China, and by early 2020 the first cases were identified in the United States. SARS-CoV-2 infections increased in the US causing many states to implement stay-at-home orders and additional safety precautions to mitigate potential outbreaks. As policies changed throughout the pandemic and restrictions lifted, there was an increase in demand for COVID-19 testing which was costly, difficult to obtain, or had long turn-around times. Some academic institutions, including Boston University (BU), created an on-campus COVID-19 screening protocol as part of a plan for the safe return of students, faculty, and staff to campus with the option for in-person classes. At BU, we put together an automated high-throughput clinical testing laboratory with the capacity to run 45,000 individual tests weekly by Fall of 2020, with a purpose-built clinical testing laboratory, a multiplexed reverse transcription PCR (RT-qPCR) test, robotic instrumentation, and trained staff. There were many challenges including supply chain issues for personal protective equipment and testing materials in addition to equipment that were in high demand. The BU Clinical Testing Laboratory (CTL) was operational at the start of Fall 2020 and performed over 1 million SARS-CoV-2 PCR tests during the 2020-2021 academic year.Boston UniversityPublished versio

Oligosaccharide substrate preferences of human extracellular sulfatase Sulf2 using liquid chromatography-mass spectrometry based glycomics approaches.

Sulfs are extracellular endosulfatases that selectively remove the 6-O-sulfate groups from cell surface heparan sulfate (HS) chain. By altering the sulfation at these particular sites, Sulfs function to remodel HS chains. As a result of the remodeling activity, HSulf2 regulates a multitude of cell-signaling events that depend on interactions between proteins and HS. Previous efforts to characterize the substrate specificity of human Sulfs (HSulfs) focused on the analysis of HS disaccharides and synthetic repeating units. In this study, we characterized the substrate preferences of human HSulf2 using HS oligosaccharides with various lengths and sulfation degrees from several naturally occurring HS sources by applying liquid chromatography mass spectrometry based glycomics methods. The results showed that HSulf2 preferentially digests highly sulfated HS oligosaccharides with zero acetyl groups and this preference is length dependent. In terms of length of oligosaccharides, HSulf2 digestion induced more sulfation decrease on DP6 (DP: degree of polymerization) compared to DP2, DP4 and DP8. In addition, the HSulf2 preferentially digests the oligosaccharide domain located at the non-reducing end (NRE) of the HS and heparin chain. In addition, the HSulf2 digestion products were altered only for specific isomers. HSulf2 treated NRE oligosaccharides also showed greater decrease in cell proliferation than those from internal domains of the HS chain. After further chromatographic separation, we identified the three most preferred unsaturated hexasaccharide for HSulf2

Extracted ion chromatograms (EICs) of -4 charged [1], [2], [3], [1,6] and [1], [2], [3], [1,7] before and after HSulf2 digestion from SEC-MS.

<p>EICs from three LC-MS runs were overlaid to the same scale and the double arrow shows the peak area that changed most upon HSulf2 digestion.</p

Composition profiling of the peaks from SAX separation with most abundance changes after HSulf2 digestion.

<p>Peaks <b>a∼e</b> represent chromatographic peaks from <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0105143#pone-0105143-g005" target="_blank">Fig. 5</a>.</p

UV chromatograms of SAX separation of HS DP6 for control and HSulf2 digestion.

<p>Peak a∼e indicates major fractions that change upon Sulf digestion.</p

Dose-response plotting of BaF32 cell proliferation assays of two different HS DP6 sources upon HSulf2 digestion.

<p>A. HS DP6 from lyase III digestion. B. HS DP6 from lyase I digestion.</p

Changes of sulfate degree before and after HSulf2 digestion of HS oligosaccharides.

<p>A. HS DP6 sulfated changes for 0 Ac, 1 Ac and 2 Ac series. B. HS DP6 sulfated changes for 0 Ac, 1 Ac and 2 Ac series. Left Y-axis denotes the sulfation degree represented as the average sulfate number per disaccharide. Right Y-axis denotes the decrease in sulfate degree after HSulf2 digestion in percentage. Data significance was calculated by two-tailed Student's <i>t</i> test, with ** denoting p<0.01 and * denoting p<0.05. NS means difference not significant.</p

Towards an HIV cure: a global scientific strategy

Given the limitations of antiretroviral therapy and recent advances in our understanding of HIV persistence during effective treatment, there is a growing recognition that a cure for HIV infection is both needed and feasible. The International AIDS Society convened a group of international experts to develop a scientific strategy for research towards an HIV cure. Several priorities for basic, translational and clinical research were identified. This Opinion article summarizes the group's recommended key goals for the international communit