The devil is in the details: emerging insights on the relevance of wastewater surveillance for SARS-CoV-2 to public health

The severe health consequences and global spread of the COVID-19 pandemic have necessitated the rapid development of surveillance programs to inform public health responses. Efforts to support surveillance capacity have included an unprecedented global research response into the use of genetic signals of SARS-CoV-2 in wastewater following the initial demonstration of the virus' detectability in wastewater in early 2020. The confirmation of fecal shedding of SARS-CoV-2 from asymptomatic, infected and recovering individuals further supports the potential for wastewater analysis to augment public health conventional surveillance techniques based on clinical testing of symptomatic individuals. We have reviewed possible capabilities projected for wastewater surveillance to support pandemic management, including independent, objective and cost-effective data generation that complements and addresses attendant limitations of clinical surveillance, early detection (i.e., prior to clinical reporting) of infection, estimation of disease prevalence, tracking of trends as possible indicators of success or failure of public health measures (mask mandates, lockdowns, vaccination, etc.), informing and engaging the public about pandemic trends, an application within sewer networks to identify infection hotspots, monitoring for presence or changes in infections from institutions (e.g., long-term care facilities, prisons, educational institutions and vulnerable industrial plants) and tracking of appearance/progression of viral variants of concern.

Arthroscopic Resection of Humeral Enchondroma

Enchondromas are common benign bone tumors. They are often found incidentally and usually do not require any treatment other than clinical surveillance. Signs of lesion progression or nontolerable pain are criteria for surgical resection. We present a case of a 44-year-old woman with long-lasting shoulder pain, diagnosed with enchondroma. Imaging studies showed an enchondroma near the great tuberosity. Shoulder arthroscopy made it possible to diagnose and treat an SLAP lesion as well as to resect by curettage and shaving through the subacromial lateral portal. One year after surgery, the patient had significant pain relief and improved shoulder function. Shoulder arthroscopy seems to be a safe and suitable technique for resection of small-sized humeral enchondromas, with the advantage of allowing to treat other possible concomitant shoulder pathologies.

Primary solitary fibrous tumor of the thyroid gland: A review starting from a case report

Primary solitary fibrous tumor (SFT) of the thyroid gland is a rare mesenchymal tumor with fibroblastic differentiation, ramified, thin-walled, enlarged (staghorn) vessels and specific NAB2-STAT6 gene fusion, which is more commonly found in pleura and peritoneum. This neoplasm can be located in a variety of anatomical sites outside pleura and peritoneum including bone, visceral organs and soft tissues, head and neck examples representing only 10-15% of the extra-pleural and extra-peritoneal tumors. Diagnosing this entity can be difficult, especially in thyroid gland, mainly because of the rarity of this neoplasm, but presence of characteristic microscopic features together with positivity for STAT6 and CD34 can confirm the diagnosis and exclude other differential diagnosis. Information about the diagnosis and treatment options of thyroid SFTs is limited but almost all primary thyroid SFTs have a good prognosis and indolent clinical course. Clinical surveillance is still necessary because some SFTs can be aggressive. Raising awareness regarding extra-pleural and extra-peritoneal location of this tumor in endocrine organs can help to better manage these patients. We report the case of a 34-year-old female with primary SFT of the thyroid gland. Additionally, we review the literature for the main clinical, paraclinical and pathological features of this neoplasm.

Detection of Pneumocystis jirovecii and Toxoplasma gondii in patients with lung infections by a duplex qPCR assay

Pneumocystis pneumonia (PCP) and pulmonary toxoplasmosis (PT) are caused by Pneumocystis jirovecii and Toxoplasma gondii. The clinical symptoms and imaging of PCP and PT are indistinguishable. A duplex qPCR was developed to differentiate between these two pathogens. In testing 92 clinical samples to validate the performance of this method for P. jirovecii detection, it identified 31 positive samples for P. jirovecii infection, consistent with clinical diagnosis. Among the remainder of the 61 clinical samples with suspected PCP, yet showing as negative by the conventional PCR diagnosis approach, 6 of them proved positive using our new assay. Our new approach also produced similar results in identification of T. gondii infections, giving a result of 2 positive and 20 negative in clinical samples. An investigation was undertaken on the prevalence of P. jirovecii and T. gondii infections using 113 samples from lung infection patients. 9% (10/113) were shown to be positive with infections of P. jirovecii, 2% with T. gondii (2/113) and 5% (6/113) were co-infected with both pathogens. Although this duplex qPCR can detect individual P. jirovecii and T. gondii infection, and co-infection of both pathogens, further large-scale investigations are needed to validate its performance, especially in T. gondii detection. Our assay provides a rapid and accurate tool for PCP and PT diagnosis in immunocompromised population and clinical surveillance of these infections in patients with no immune defects.

Combining community wastewater genomic surveillance with state clinical surveillance: A framework for SARS-CoV-2 public health practice

Study objectiveTo garner a framework for combining community wastewater surveillance with state clinical surveillance that influence confirmation of SARS-CoV-2 variants within the community, and recommend how the flow of such research evidence could be expanded and employed for public health response.Design, setting, and participantsThis work involved analyzing wastewater samples collected weekly from 17 geographically resolved locations in Louisville/Jefferson County, Kentucky from February 10 to November 29, 2021. Genomic surveillance and RT-qPCR platforms were used as screening to identify SARS-CoV-2 in wastewater, and state clinical surveillance was used for confirmation.Main resultsThe results demonstrate increased epidemiological value of combining community wastewater genomic surveillance and RT-qPCR with conventional case auditing methods. The spatial scale and temporal frequency of wastewater sampling provides promising sensitivity and specificity to be useful to gain public health screening insights about community emergence, seeding, and spread.ConclusionsBetter national surveillance systems are needed for future pathogens and variants, and wastewater-based genomic surveillance represents opportune coupling. This paper presents current evidence that complementary wastewater and clinical testing is enhanced cost-effectively when linked; making a strong case for a joint public health framework. The findings suggest significant potential for rapid progress to be made in extending this work to consider pathogens of interest as a whole within wastewater, which could be examined in either a targeted fashion as we currently do with SARS-CoV-2 or in terms of a global monitoring of all pathogens found, and developing evidence based public health practice to best support community health.Thumbnail BoxWhat is already known on this subject?The primary approach for the genomic surveillance of SARS-CoV-2 relies on the sequencing of clinical COVID-19 samples. Variants of SARS-CoV-2 can also be tracked in community wastewater.What this study adds?We propose that, for comprehensive community surveillance, the first line of community pathogen screening should involve geographically-resolved wastewater samples collected at a regular frequency and employ both Next Generation Sequencing (NGS) and RT-qPCR. These results could then be compared with state clinical surveillance. This framework is a more comprehensive and cost-effective approach for surveillance in practice to catch community emergence, seeding, and spread.Policy implicationsOur results present a framework that could support the implementation of better surveillance policies directed to solve future community pathogen and variant detection. We anticipate this work can help public health officials implement rational community sampling schemes and develop sensible coordination with other clinical surveillance. The utility of this for COVID-19 extends to many other infectious disease models and other public health hazards such as toxic exposures.

LuNER: Multiplexed SARS-CoV-2 detection in clinical swab and wastewater samples

Clinical and surveillance testing for the SARS-CoV-2 virus relies overwhelmingly on RT-qPCR-based diagnostics, yet several popular assays require 2–3 separate reactions or rely on detection of a single viral target, which adds significant time, cost, and risk of false-negative results. Furthermore, multiplexed RT-qPCR tests that detect at least two SARS-CoV-2 genes in a single reaction are typically not affordable for large scale clinical surveillance or adaptable to multiple PCR machines and plate layouts. We developed a RT-qPCR assay using the Luna Probe Universal One-Step RT-qPCR master mix with publicly available primers and probes to detect SARS-CoV-2 N gene, E gene, and human RNase P (LuNER) to address these shortcomings and meet the testing demands of a university campus and the local community. This cost-effective test is compatible with BioRad or Applied Biosystems qPCR machines, in 96 and 384-well formats, with or without sample pooling, and has a detection sensitivity suitable for both clinical reporting and wastewater surveillance efforts.

Australian National Enterovirus Reference Laboratory annual report, 2020

Australia monitors its polio-free status by conducting surveillance for cases of acute flaccid paralysis (AFP) in children less than 15 years of age, as recommended by the World Health Organization (WHO). Cases of AFP in children are notified to the Australian Paediatric Surveillance Unit or the Paediatric Active Enhanced Disease Surveillance System and faecal specimens are referred for virological investigation to the National Enterovirus Reference Laboratory. In 2020, no cases of poliomyelitis were reported from clinical surveillance; Australia reported 1.09 non-polio AFP cases per 100,000 children, thereby meeting the WHO’s performance criterion for a sensitive surveillance system. The non-polio enteroviruses coxsackievirus A10 and coxsackievirus A16 were identified from clinical specimens collected from AFP cases. Australia also performs enterovirus surveillance and environmental surveillance to complement the clinical system focussed on children. In 2020, there were 140 cases of wild poliovirus reported from the two remaining endemic countries: Afghanistan and Pakistan. Another 28 countries reported cases of circulating vaccine-derived poliovirus.