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Quantitative, reverse transcription, polymerase chain reaction (qRT-PCR) is facilitated by leveraging droplet microfluidic (DMF) system, which due to its precision dispensing and sample handling capabilities at microliter and lowe...
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Quantitative, reverse transcription, polymerase chain reaction (qRT-PCR) is facilitated by leveraging droplet microfluidic (DMF) system, which due to its precision dispensing and sample handling capabilities at microliter and lower volumes has emerged as a popular method for miniaturization of the PCR platform. This work substantially improves and extends the functional capabilities of our previously demonstrated single qRT-PCR micro-chip, which utilized a combination of electrostatic and electrowetting droplet actuation. In the reported work we illustrate a spatially multiplexed micro-device that is capable of conducting up to eight parallel, real-time PCR reactions per usage, with adjustable control on the PCR thermal cycling parameters (both process time and temperature set-points). This micro-device has been utilized to detect and quantify the presence of two clinically relevant respiratory viruses, Influenza A and Influenza B, in human samples (nasopharyngeal swabs, throat swabs). The device performed accurate detection and quantification of the two respiratory viruses, over several orders of RNA copy counts, in unknown (blind) panels of extracted patient samples with acceptably high PCR efficiency (>94%). The multi-stage qRT-PCR assays on eight panel patient samples were accomplished within 35–40 min, with a detection limit for the target Influenza virus RNAs estimated to be less than 10 RNA copies per reaction.
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SARS-CoV-2 variants of concern (VOCs) have emerged as a global threat to the COVID-19 pandemic response. We implemented a combined approach to quickly detect known VOCs while continuously monitoring for evolving mutations of the v...
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SARS-CoV-2 variants of concern (VOCs) have emerged as a global threat to the COVID-19 pandemic response. We implemented a combined approach to quickly detect known VOCs while continuously monitoring for evolving mutations of the virus. To rapidly detect VOCs, two real-time reverse transcriptase PCR assays were designed and implemented, targeting the spike gene H69/V70 deletion and the N501Y mutation. The H69/V70 deletion and N501Y mutation assays demonstrated accuracies of 98.3% (95% CI 93.8 to 99.8) and 100% (95% CI 96.8 to 100), limits of detection of 1,089 and 294 copies/ml, and percent coefficients of variation of 0.08 to 1.16% and 0 to 2.72% for the two gene targets, respectively. No cross-reactivity with common respiratory pathogens was observed with either assay. Implementation of these tests allowed the swift escalation in testing for VOCs from 2.2% to ～100% of all SARS-CoV-2-positive samples over 12 January to 9 February 2021, and resulted in the detection of a rapid rise of B.1.1.7 cases within the province of Alberta, Canada. A prospective comparison of the VOC assays to genome sequencing for the detection of B.1.1.7, combined detection of P.1 and B.1.351, and wild-type (i.e., non-VOC) lineages showed sensitivities of 98.2 to 100%, specificities of 98.9 to 100%, positive predictive values of 76.9% to 100%, and negative predictive values of 96 to 100%. Variant screening results inform sampling strategies for regular surveillance by genome sequencing, thus allowing rapid identification of known VOCs while continuously monitoring the evolution of SARS-CoV-2 in the province. IMPORTANCE Different strains, or variants, of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, the virus that causes COVID-19) have emerged that have higher levels of transmission, less susceptibility to our immune response, and possibly cause more severe disease than previous strains of the virus. Rapid detection of these variants of concern is important to help contain them and prevent them from spreading widely within the population. This study describes two newly developed tests that are able to identify and differentiate the variants of concern from regular strains of SARS-CoV-2. These tests are faster and simpler than the main, gold standard method of identifying variants of concern (genome sequencing). These tests also demonstrated a high correlation with genome sequencing and allowed for the rapid and accurate detection of the rise of B.1.1.7 (one of the variants of concern) in the province of Alberta, Canada.
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The emergence of norovirus genotype GII.4 variants has been associated with gastroenteritis pandemics worldwide, prompting molecular surveillance for early detection of novel strains. In this study, we aimed to analyze the outbrea...
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The emergence of norovirus genotype GII.4 variants has been associated with gastroenteritis pandemics worldwide, prompting molecular surveillance for early detection of novel strains. In this study, we aimed to analyze the outbreak activity of norovirus and characterize the norovirus strains circulating in Alberta between July 2012 and February 2018. Stool samples from gastroenteritis outbreaks in Alberta were tested for norovirus at the Provincial Laboratory for Public Health using a multiplex real time-RT PCR assay. The ORF1 and ORF2-genotypes of norovirus positive samples were assigned based on phylogenetic analyses of partial polymerase and capsid sequences, respectively. A total of 530 norovirus outbreaks were identified. During July 2012 and June 2017 there was a gradual decrease in the annual number of GII.4 outbreaks, however, outbreak numbers increased from June 2017-February 2018. Four novel strains emerged: GII.17 Kawasaki in July 2014-June 2015, GII.P16/GII.4 Sydney in July 2015-June 2016, GII.P16/GII.2 and GII.P4 New Orleans/GII.4 Sydney in July 2016-June 2017. GII.Pe/GII.4 Sydney was the single predominant strain responsible for the majority (over 50%) of all norovirus outbreaks up to June 2015. Between June 2017 and February 2018, GII.P16/GII.4 Sydney was the leading strain causing 63% of all norovirus outbreaks. GII.4 stands as the predominant capsid genotype causing a large majority of the norovirus outbreaks in early 2018. An increase in genotype diversity was observed in the last years, characterized by a high circulation of non-GII.4 strains and GII.4 recombinants.
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Background: Children who are neurodiverse have traditionally been segregated from their peers in community-based programs, despite evidence of health benefits of inclusive education. Objectives: This community-initiated project ai...
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Background: Children who are neurodiverse have traditionally been segregated from their peers in community-based programs, despite evidence of health benefits of inclusive education. Objectives: This community-initiated project aims to explore barriers and facilitators to inclusive aquatics programming for children with developmental and/or mental health challenges. Methods: Using a participatory-action research methodology, semi-structured interviews and focus groups were conducted with 14 participants from various stakeholder groups, including parents of children who are neurodiverse, helping professionals, and community programmers. Results: Participants described unique definitions of inclusion, from integration with neurotypical peers, to individualized goal-setting and achievement. Major facilitators include adequate resources, flexibility around accommodations, and motivated staff. Major barriers include social stigma, financial limitations, and lack of communication between caregivers and service providers. Conclusions: Participants felt strongly about the need to improve inclusion practices within aquatics—and other community-based—programs. Increased collaboration between families, community programmers, and helping professionals can foster better inclusion and outcomes for children who are neurodiverse. By incorporating various perspectives into the design of future programs, program administrators can ensure more equitable access such that all children are able to participate.
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Abstract: In Hong Kong, the evidence for cognitive-training programs in fighting against memory complaints is lacking. This study aimed to evaluate the effectiveness of the Active Mind cognitive-training program in improving the c...
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Abstract: In Hong Kong, the evidence for cognitive-training programs in fighting against memory complaints is lacking. This study aimed to evaluate the effectiveness of the Active Mind cognitive-training program in improving the cognitive function and quality of life (QoL) for local community-dwelling Chinese older adults. A total of 200 subjects were recruited from 20 different district elderly community centers (DECCs). Centers were randomly assigned into either the intervention group or control group. The intervention group underwent eight 1-hour sessions of cognitive training, while the control group were included in the usual group activities provided by the DECCs. Standardized neuropsychological tests (the Chinese version of Mattis Dementia Rating Scale [CDRS] and the Cantonese version of the Mini-Mental State Examination) and the QoL questionnaire SF12 were used to assess participants' cognitive function and QoL before and after the trial. A total of 176 subjects completed the study. The intervention group showed greater improvement in the cognitive function measured by total CDRS score (treatment: 12.24 ± 11.57 vs control: 4.37 ± 7.99; P < 0.001) and QoL measured by total SF12 score (treatment: 7.82 ± 13.19 vs control: 3.18 ± 11.61; P = 0.014). Subjects with lower education level were associated with better cognitive response to the cognitive-training program. The current findings indicated that the Active Mind cognitive-training program was effective in improving the cognitive function and QoL for community-dwelling Chinese older adults in Hong Kong.
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AbstractBackgroundInfluenza C virus can cause both upper and lower respiratory tract infections and has been reported to be prevalent in children. However, these infections have been under-diagnosed, and epidemiological data avail...
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AbstractBackgroundInfluenza C virus can cause both upper and lower respiratory tract infections and has been reported to be prevalent in children. However, these infections have been under-diagnosed, and epidemiological data available are limited due to the lack of convenient detection assays.ObjectiveDesign and validate a real-time reverse-transcriptase PCR (rt RT-PCR) assay for the detection of influenza C.Study designRespiratory samples from two primary settings, namely, children who were hospitalized or seen in the emergency department, and respiratory outbreaks for which no other viral etiology was found were used for the detection of influenza C.Results and ConclusionsThe assay was sensitive and specific for the detection of influenza C. Eleven of 474 (2·32%) patients, all less than 10 years of age, were positive for influenza C. The strains clustered into two lineages, namely C/Kanagawa and C/Sao Paulo, based upon sequencing of the hemagglutinin-esterase gene. Epidemiological data showed that a higher proportion of influenza C infections occur in younger children and during the winter months. This is the first report of the detection of influenza C in Alberta, Canada, and suggests that the detection of this virus should be included in respiratory virus testing panels.
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The performance characteristics of the RealART and Molecular Beacons assays were compared with those of the Digene Hybrid Capture II assay (ultrasensitive). The results of the RealART and Digene Hybrid assays were related (r = 0.9...
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The performance characteristics of the RealART and Molecular Beacons assays were compared with those of the Digene Hybrid Capture II assay (ultrasensitive). The results of the RealART and Digene Hybrid assays were related (r = 0.94; P < 0.001) and diverged by 2 orders of magnitude. The RealART assay can be used to effectively monitor serum hepatitis B virus DNA levels.
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Food-dependent exercise-induced anaphylaxis is a rare and underdiagnosed condition in which anaphylactic reactions present only in the setting of exercise within a few hours following (or rarely, immediately before) food intake. T...
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Food-dependent exercise-induced anaphylaxis is a rare and underdiagnosed condition in which anaphylactic reactions present only in the setting of exercise within a few hours following (or rarely, immediately before) food intake. This case describes a 20-year old female with severe anaphylaxis developing during a run shortly after ingesting a meal containing shrimp. With acute epinephrine treatment, allergy workup, and education regarding separation of shrimp and exercise, this patient was able to resume her normal diet and exercise regimen without further harm. We hope this case raises awareness of this condition which may be overlooked by many clinicians due to lack of familiarity with the condition.
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