Simple tape-stripping method for highly reliable and quantitative analysis of skin microbiome.

The composition of human skin microbiome profoundly impacts host skin health and disease. However, the relationship between skin homeostasis or the development of skin diseases and daily changes in skin microbial composition is poorly understood. Longitudinal samplings at more frequent intervals would address this issue, while conventional sampling methods have technical difficulties, leading to limitations in sampling opportunities. Here, we developed a simple and stable tape-stripping method regardless of the operator's skill. Our method enables skin microbial sampling within 30 seconds and taking multiple skin microbial samples from the same body site. The amount of microbial DNA among multiple sampling sites could be measured within 13.5%. The sequencing results of multiple sampling showed high consistency, Pearson's correlation coefficient between multiple samples of 0.98. Furthermore, these results were comparable to those collected by the conventional swabbing method. These results demonstrate that our tape-stripping method enables simple microbiome collection and highly reliable quantitative skin microbiome analysis. These features of our method would lead to a further understanding of skin disease development or diagnosis of skin conditions in clinical research by increasing the opportunities for microbial sampling.

Self-swabbing versus assisted swabbing for viral detection by qRT-PCR: the experience from SARS-CoV-2 based on a meta-analysis of six prospectively designed evaluations conducted in a UK setting.

PURPOSE: In April 2020, the UK Government implemented NHS Test and Trace to provide SARS-CoV-2 quantitative reverse transcription polymerase chain reaction (qRT-PCR) testing for the public, with nose-and-throat swabbing for samples performed by trained staff. Self-swabbing (SS) would allow rapid scale-up of testing capacity and access. Six studies were undertaken to determine whether SS was as effective for detecting SARS-CoV-2 as swabbing performed by trained staff. METHODS: Six prospective studies were conducted between April-October 2020, using six swab/media combinations. Differences between assisted swabbing (AS) and SS were evaluated for concordance, positivity, sensitivity, cycle threshold (Ct) values and void rates. Statistical analysis was performed using 95% confidence intervals (CIs), paired t-tests and model-based methods. RESULTS: Overall, 3,253 individuals were recruited (median age 37 years, 49% female), with 2,933 having valid paired qRT-PCR results. Pooled concordance rate was 98% (95% CI: 96%, 99%). Positivity rate differences for SS (8.1%) and AS (8.4%) and differences in pooled sensitivities between SS (86%; 95% CI: 78%, 92%) and AS (91%; 95% CI: 78%, 96%) were nonsignificant. Both types of swabbing led to pooled void rates below 2% and strongly correlated Ct values. Age, sex and previous swabbing experience did not have a significant impact on concordance or sensitivity. CONCLUSION: The UK adopted a policy to promote self-testing for SARS-CoV-2 based on data demonstrating equivalence of SS versus AS. Positive outcomes with SS are likely generalisable to testing for other respiratory pathogens, and we consider self-sampling and self-testing essential for future pandemic preparedness.

eProbe: Sampling of Environmental DNA within Tree Canopies with Drones.

Environmental DNA (eDNA) analysis is a powerful tool for studying biodiversity in forests and tree canopies. However, collecting representative eDNA samples from these high and complex environments remains challenging. Traditional methods, such as surface swabbing or tree rolling, are labor-intensive and require significant effort to achieve adequate coverage. This study proposes a novel approach for unmanned aerial vehicles (UAVs) to collect eDNA within tree canopies by using a surface swabbing technique. The method involves lowering a probe from a hovering UAV into the canopy and collecting eDNA as it descends and ascends through branches and leaves. To achieve this, a custom-designed robotic system was developed featuring a winch and a probe for eDNA collection. The design of the probe was optimized, and a control logic for the winch was developed to reduce the risk of entanglement while ensuring sufficient interaction force to facilitate transfer of eDNA onto the probe. The effectiveness of this method was demonstrated during the XPRIZE Rainforest Semi-Finals as 10 eDNA samples were collected from the rainforest canopy, and a total of 152 molecular operational taxonomic units (MOTUs) were identified using eDNA metabarcoding. We further investigate how the number of probe interactions with vegetation, the penetration depth, and the sampling duration influence the DNA concentration and community composition of the samples.

The Effect of Stress Ball Use Applied by Emergency Nurses During Swabbing Procedure on the Pain and Fear Levels of Children Admitted to the Pediatric Emergency Service With the Suspicion of COVID-19: A Randomized Controlled Trial.

INTRODUCTION: This study aimed to determine the effect of stress ball use during the swabbing procedure on the pain and fear levels of children admitted to the pediatric emergency department with the suspicion of coronavirus disease 2019. Children with suspected coronavirus disease 2019 were recruited by convenience sampling from the pediatric emergency department of a university hospital in a city in Turkey. METHODS: This study used a random controlled experimental design and had a calculated sample size of 60. There were 30 participants in both the control and experimental groups. The stress ball intervention was applied to the children in the experimental group during the swabbing process, and no intervention was made to the children in the control group during the procedure. The pain and fear levels of the children in the control and experimental groups were measured during the swabbing process. "Descriptive Characteristics Form for Parents and Children," "Wong-Baker Faces Pain Rating Scale," "Children's Fear Scale," and "Stress Ball" were used in data collection. Chi-square, Mann-Whitney U, and Friedman tests were used in the analysis. RESULTS: Although there was no statistically significant difference between the groups in terms of pain and fear level mean scores before the procedure, a statistically significant difference was found between the groups during and after the procedures (P < .05). DISCUSSION: Giving a stress ball to children aged 4 to 10 years during the swabbing procedure was determined to reduce the pain and fear levels during and after the procedures. It is recommended that stress ball use be applied during the swabbing procedure for children.

RNAseq analysis of olfactory neuroepithelium cytological samples in individuals with Down syndrome compared to euploid controls: a pilot study.

Down syndrome is a common genetic disorder caused by partial or complete triplication of chromosome 21. This syndrome shows an overall and progressive impairment of olfactory function, detected early in adulthood. The olfactory neuronal cells are located in the nasal olfactory mucosa and represent the first sensory neurons of the olfactory pathway. Herein, we applied the olfactory swabbing procedure to allow a gentle collection of olfactory epithelial cells in seven individuals with Down syndrome and in ten euploid controls. The aim of this research was to investigate the peripheral gene expression pattern in olfactory epithelial cells through RNAseq analysis. Validated tests (Sniffin' Sticks Extended test) were used to assess olfactory function. Olfactory scores were correlated with RNAseq results and cognitive scores (Vineland II and Leiter scales). All Down syndrome individuals showed both olfactory deficit and intellectual disability. Down syndrome individuals and euploid controls exhibited clear expression differences in genes located in and outside the chromosome 21. In addition, a significant correlation was found between olfactory test scores and gene expression, while a non-significant correlation emerged between olfactory and cognitive scores. This first preliminary step gives new insights into the Down syndrome olfactory system research, starting from the olfactory neuroepithelium, the first cellular step on the olfactory way.

Internet Tool to Support Self-Assessment and Self-Swabbing of Sore Throat: Development and Feasibility Study.

BACKGROUND: Sore throat is a common problem and a common reason for the overuse of antibiotics. A web-based tool that helps people assess their sore throat, through the use of clinical prediction rules, taking throat swabs or saliva samples, and taking throat photographs, has the potential to improve self-management and help identify those who are the most and least likely to benefit from antibiotics. OBJECTIVE: We aimed to develop a web-based tool to help patients and parents or carers self-assess sore throat symptoms and take throat photographs, swabs, and saliva samples for diagnostic testing. We then explored the acceptability and feasibility of using the tool in adults and children with sore throats. METHODS: We used the Person-Based Approach to develop a web-based tool and then recruited adults and children with sore throats who participated in this study by attending general practices or through social media advertising. Participants self-assessed the presence of FeverPAIN and Centor score criteria and attempted to photograph their throat and take throat swabs and saliva tests. Study processes were observed via video call, and participants were interviewed about their views on using the web-based tool. Self-assessed throat inflammation and pus were compared to clinician evaluation of patients' throat photographs. RESULTS: A total of 45 participants (33 adults and 12 children) were recruited. Of these, 35 (78%) and 32 (71%) participants completed all scoring elements for FeverPAIN and Centor scores, respectively, and most (30/45, 67%) of them reported finding self-assessment relatively easy. No valid response was provided for swollen lymph nodes, throat inflammation, and pus on the throat by 11 (24%), 9 (20%), and 13 (29%) participants respectively. A total of 18 (40%) participants provided a throat photograph of adequate quality for clinical assessment. Patient assessment of inflammation had a sensitivity of 100% (3/3) and specificity of 47% (7/15) compared with the clinician-assessed photographs. For pus on the throat, the sensitivity was 100% (3/3) and the specificity was 71% (10/14). A total of 89% (40/45), 93% (42/45), 89% (40/45), and 80% (30/45) of participants provided analyzable bacterial swabs, viral swabs, saliva sponges, and saliva drool samples, respectively. Participants were generally happy and confident in providing samples, with saliva samples rated as slightly more acceptable than swab samples. CONCLUSIONS: Most adult and parent participants were able to use a web-based intervention to assess the clinical features of throat infections and generate scores using clinical prediction rules. However, some had difficulties assessing clinical signs, such as lymph nodes, throat pus, and inflammation, and scores were assessed as sensitive but not specific. Many participants had problems taking photographs of adequate quality, but most were able to take throat swabs and saliva samples.

Human skin biofilm model: translational impact on swabbing and debridement.

OBJECTIVE: Wound biofilms are one of the greatest challenges in the therapy of hard-to-heal (chronic) wounds, as potent antimicrobial substances fail to eradicate bacteria within short incubation periods. Preclinical investigations using novel model systems that closely mimic the human wound environment and wound biofilm are required to identify new and effective therapeutic options. This study aims to identify bacterial colonisation patterns that are relevant for diagnosis and therapy. METHOD: In this study, a recently established human plasma biofilm model (hpBIOM) was incorporated into a wound within human dermal resectates after abdominoplasty. The interaction of the biofilm-forming bacteria meticillin-resistant Staphylococcus aureus (MRSA) and Pseudomonas aeruginosa with the skin cells was investigated. Possible effects on wound healing processes in correlation with the persistence of the biofilm in the wound environment were analysed in patients with leg ulcers of different aetiologies and biofilm burden. RESULTS: Using haematoxylin and eosin staining, species-dependent infiltration modes of the bacteria into the wound tissue were determined for the pathogens MRSA and Pseudomonas aeruginosa. The spreading behaviour correlated with clinical observations of the spatial distributions of the bacteria. In particular, the clinically prominent Pseudomonas aeruginosa-specific distension of the wound margin was identified as epidermolysis due to persistent infiltration. CONCLUSION: The hpBIOM applied in this study represents a potential tool for preclinical analyses dealing with approval processes for new antimicrobial applications. In terms of clinical practice, a microbiological swabbing technique including the wound margin should be routinely applied to prevent wound exacerbation.

Alpha-synuclein seeds in olfactory mucosa of patients with isolated REM sleep behaviour disorder.

Isolated REM sleep behaviour disorder (RBD) is an early-stage α-synucleinopathy in most, if not all, affected subjects. Detection of pathological α-synuclein in peripheral tissues of patients with isolated RBD may identify those progressing to Parkinson's disease, dementia with Lewy bodies or multiple system atrophy, with the ultimate goal of testing preventive therapies. Real-time quaking-induced conversion (RT-QuIC) provided evidence of α-synuclein seeding activity in CSF and olfactory mucosa of patients with α-synucleinopathies. The aim of this study was to explore RT-QuIC detection of α-synuclein aggregates in olfactory mucosa of a large cohort of subjects with isolated RBD compared to patients with Parkinson's disease and control subjects. This cross-sectional case-control study was performed at the Medical University of Innsbruck, Austria, the Hospital Clinic de Barcelona, Spain, and the University of Verona, Italy. Olfactory mucosa samples obtained by nasal swab in 63 patients with isolated RBD, 41 matched Parkinson's disease patients and 59 matched control subjects were analysed by α-synuclein RT-QuIC in a blinded fashion at the University of Verona, Italy. Median age of patients with isolated RBD was 70 years, 85.7% were male. All participants were tested for smell, autonomic, cognitive and motor functions. Olfactory mucosa was α-synuclein RT-QuIC positive in 44.4% isolated RBD patients, 46.3% Parkinson's disease patients and 10.2% control subjects. While the sensitivity for isolated RBD plus Parkinson's disease versus controls was 45.2%, specificity was high (89.8%). Among isolated RBD patients with positive α-synuclein RT-QuIC, 78.6% had olfactory dysfunction compared to 21.4% with negative α-synuclein RT-QuIC (P < 0.001). The extent of olfactory dysfunction was more severe in isolated RBD patients positive than negative for olfactory mucosa a-synuclein RT-QuIC (P < 0.001). We provide evidence that the α-synuclein RT-QuIC assay enables the molecular detection of neuronal α-synuclein aggregates in olfactory mucosa of patients with isolated RBD and Parkinson's disease. Although the overall sensitivity was moderate in this study, nasal swabbing is attractive as a simple, non-invasive test and might be useful as part of a screening battery to identify subjects in the prodromal stages of α-synucleinopathies. Further studies are needed to enhance sensitivity, and better understand the temporal dynamics of α-synuclein seeding in the olfactory mucosa and spreading to other brain areas during the progression from isolated RBD to overt α-synucleinopathy, as well the impact of timing, disease subgroups and sampling technique on the overall sensitivity.

Side-by-side comparison of parent vs. technician-collected respiratory swabs in low-income, multilingual, urban communities in the United States.

BACKGROUND: Home-based swabbing has not been widely used. The objective of this analysis was to compare respiratory swabs collected by mothers of 7-12-year-olds living in low-income, multilingual communities in the United States with technician collected swabs. METHODS: Retrospective data analysis of respiratory samples collected at home by mothers compared to technicians. Anterior nasal and throat specimens collected using flocked swabs were combined in dry tubes. Test was done using TaqMan array cards for viral and bacterial pathogens. Cycle threshold (Ct) values of ribonuclease P (RNP) gene were used to assess specimen quality. Ct < 40 was interpreted as a positive result. Concordance of pathogen yield from mother versus technician collected swabs were analyzed using Cohen's Kappa coefficients. Correlation analysis, paired t-test, and Wilcoxon signed-rank test for paired samples were used for RNP Ct values. RESULTS: We enrolled 36 households in Cincinnati (African American) and 44 (predominately Chinese or Latino) in Boston. In Cincinnati, eight of 32 (25%) mothers did not finish high school, and 11 (34%) had finished high school only. In Boston, 13 of 44 (30%) mothers had less than a high school diploma, 23 (52%) had finished high school only. Mother versus technician paired swabs (n = 62) had similar pathogen yield (paired t-test and Wilcoxon signed rank test p-values = 0.62 and 0.63, respectively; 95% confidence interval of the difference between the two measurements = - 0.45-0.75). Median Ct value for RNP was 22.6 (interquartile range, IQR = 2.04) for mother-collected and 22.4 (IQR = 2.39) for technician-collected swabs (p = 0.62). Agreement on pathogen yield between samples collected by mothers vs. technicians was higher for viruses than for bacterial pathogens, with high concordance for rhinovirus/enterovirus, human metapneumovirus, and adenovirus (Cohen's kappa coefficients ≥80%, p < 0.0001). For bacterial pathogens, concordance was lower to moderate, except for Chlamydia pneumoniae, for which kappa coefficient indicated perfect agreement. CONCLUSION: Mothers with a range of education levels from low-income communities were able to swab their children equally well as technicians. Home-swabbing using dry tubes, and less invasive collection procedures, could enhance respiratory disease surveillance.

Development and Application of Loop-Mediated Isothermal Amplification (LAMP) Assays for Rapid Diagnosis of the Bat White-Nose Disease Fungus Pseudogymnoascus destructans.

Pseudogymnoascus destructans (= Geomyces destructans) is a psychrophilic filamentous fungus that causes White-Nose Disease (WND; the disease associated with White-Nose Syndrome, WNS) in hibernating bats. The disease has caused considerable reductions in bat populations in the USA and Canada since 2006. Identification and detection of the pathogen in pure cultures and environmental samples is routinely based on qPCR or PCR after DNA isolation and purification. Rapid and specific direct detection of the fungus in the field would strongly improve prompt surveillance, and support control measures. Based on the genes coding for ATP citrate lyase1 (acl1) and the 28S-18S ribosomal RNA intergenic spacer (IGS) in P. destructans, two independent LAMP assays were developed for the rapid and sensitive diagnosis of the fungus. Both assays could discriminate P. destructans from 159 tested species of filamentous fungi and yeasts. Sensitivity of the assays was 2.1 picogram per reaction (pg/rxn) and 21 femtogram per reaction (fg/rxn) for the acl1 and IGS based assays, respectively. Moreover, both assays also work with spores and mycelia of P. destructans that are directly added to the master mix without prior DNA extraction. For field-diagnostics, we developed and tested a field-applicable version of the IGS-based LAMP assay. Lastly, we also developed a protocol for preparation of fungal spores and mycelia from swabs and tape liftings of contaminated surfaces or infected bats. This protocol in combination with the highly sensitive IGS-based LAMP-assay enabled sensitive detection of P. destructans from various sources.

A collaborative robot for COVID-19 oropharyngeal swabbing.

The coronavirus disease 2019 (COVID-19) outbreak has increased mortality and morbidity world-wide. Oropharyngeal swabbing is a well-known and commonly used sampling technique for COVID-19 diagnose around the world. We developed a robot to assist with COVID-19 oropharyngeal swabbing to prevent frontline clinical staff from being infected. The robot integrates a UR5 manipulator, rigid-flexible coupling (RFC) manipulator, force-sensing and control subsystem, visual subsystem and haptic device. The robot has strength in intrinsically safe and high repeat positioning accuracy. In addition, we also achieve one-dimensional constant force control in the automatic scheme (AS). Compared with the rigid sampling robot, the developed robot can perform the oropharyngeal swabbing procedure more safely and gently, reducing risk. Alternatively, a novel robot control schemes called collaborative manipulation scheme (CMS) which combines a automatic phase and teleoperation phase is proposed. At last, comparative experiments of three schemes were conducted, including CMS, AS, and teleoperation scheme (TS). The experimental results shows that CMS obtained the highest score according to the evaluation equation. CMS has the excellent performance in quality, experience and adaption. Therefore, the proposal of CMS is meaningful which is more suitable for robot-sampling.

A mobile swabbing booth to address Singapore GPs' concerns about swabber protection: human-centred design during the COVID-19 pandemic.

BACKGROUND: During the COVID-19 pandemic, the Ministry of Health asked Singapore's private general practitioners (GPs) to perform swab testing in their clinics, but some GPs had concerns about swabber protection. Our aim was to develop a swabbing booth to address these concerns. METHODS: We developed a prototype with potential GP users using a human-centred design approach and piloted it with 10 GP clinics. The pilot was then extended to 170 GP clinics around Singapore. These GPs were then surveyed on user satisfaction. RESULTS: Ninety-three GPs (54%) responded. The majority (75%) practiced in public residential estates in small practices (mean 1.95 doctors). 86% requested the booth to enhance swabber protection. 74% "would recommend" or "would strongly recommend" the booth to colleagues. 79% continue to use the booth to conduct swab tests. 92% liked that it offered swabber protection. 71% liked that the booth created a separate space for swabbing and 64% liked its ease of disinfection. 47% started swabbing only after receiving the booth and 58% said the booth was "important" or "very important" to their decision to participate in swab testing. However, 34% disliked that it took up too much space and the most frequently critiqued area was the gloves. CONCLUSION: The human-centred design approach generated a product that had high user satisfaction, addressed GPs' concerns of swabber protection and increased GPs' participation in swab testing. The booth may be useful where GPs are concerned about swabber protection and space is limited.

[A proposal for the management of health surveillance and monitoring procedures in relation to the risks posed by SARS-COV-2 to hospital and nursing home workers, based on strategies implemented in the facilities of the Fondazione Opera San Camillo]. / Proposta per la gestione delle attività di sorveglianza sanitaria e monitoraggio in relazione al rischio SARS-CoV-2 in operatori di strutture sanitarie ospedaliere e socioassistenziali sulla base dell'esperienza condotta nelle strutture della Fondazione Opera San Camillo.

SUMMARY: This paper describes how the health surveillance protocols adopted in hospitals and nursing homes have changed in response to the spread of SARS-CoV-2 and its risks. These changes concern assessments that must be performed with urgency, to detect potential cases and symptoms that may indicate contagion and to protect vulnerable and hypersensitive workers. Besides, these changes have been implemented in order to regulate how the appointed occupational physician should perform everyday tasks, particularly in regard to preventive visits, periodic visits, office visits, and visits meant to authorize one's return to the workplace. This paper recommends that both preventive visits and periodic visits should include rapid antigen swab tests to screen for SARS-CoV-2 (which, if positive, should be confirmed by molecular tests) and serologic tests to detect IgG SARS-CoV-2 antibodies. Since this phase involves great risks for healthcare facilities, this paper proposes that swab testing should be performed every month and that serologic testing should be performed every three months (depending on whether the first test detects the presence of IgG antibodies). By combining antigen and molecular swab tests, healthcare professionals can diagnose workers with SARS-CoV-2 and identify positive asymptomatic cases. Because of its costs and its long turnaround times for results, molecular swab testing is recommended only when one needs to confirm a positive antigen swab test result. Serologic testing should be used alongside swab testing, to acquire data concerning the immunity of workers.

Oral cavity swabbing for diagnosis of group a Streptococcus: a prospective study.

BACKGROUND: Throat pain is a common complaint in the ambulatory setting. Diagnosis of group A Streptococcus is made with a culture, molecular test or a rapid antigen detection test from the tonsils or the posterior pharyngeal wall, while other areas of the oral cavity are considered unacceptable. The purpose of the study is to compare cultures from the tonsils or posterior pharyngeal wall (throat) with cultures from the oral cavity (mouth). METHODS: A prospective study conducted in ambulatory care. Eleven family physicians collected 2 swabs (throat and mouth) from 200 consecutive patients who complaint about throat pain. Inclusion criteria were throat pain and Centor Criteria > 2. Exclusion criteria were tonsillectomy and age (< 3 or > 65 years old). Participants were later divided into two groups - pediatrics (3-18 years old) and adults (19-65 year old). Sensitivity and specificity of mouth culture were calculated, with throat culture considered the reference gold standard. RESULTS: Between November 2017 and March 2019, 200 swabs were collected (101 adults and 99 children). In the adult group sensitivity of mouth culture was 72.1% (95% Confidence Interval [CI] 59.9-82.3%) and specificity was 100% (95% CI 92.7-89.4%-100%). In the pediatric group sensitivity of mouth culture was 78.3% (95% CI 65.8-87.9%) and specificity was 100% (95% CI 92.5-100%). CONCLUSION: Our study demonstrated higher sensitivity of mouth culture for GAS than previously published. This finding suggests that areas of the oral cavity that were considered as unacceptable sites for culture of GAS pharyngitis may be considered as acceptable swabbing sites. TRIAL REGISTRATION: Trial registration: ClinicalTrials.gov, ID NCT03137823. Registered 3 May 2017.

A 3D spongy flexible nanosheet array for on-site recyclable swabbing extraction and subsequent SERS analysis of thiram.

A sponge inspired three dimensional flexible aluminum foil based ZnO nanosheet array substrate is described for use in real-world surface enhanced Raman spectroscopic detection. Gold and silver nanoparticles were employed to form numerous hot spots on uniformly grown ZnO nanosheets on the substrate. This flexible spongy substrate can extract analytes (such as the fungicide thiram) from various complex sample surfaces by physical swabbing. Specifically, this substrate was applied to detect thiram on the surface of fruits and vegetables. Non-destructive recycling detection with a relative standard deviation of 6.1% was accomplished by monitoring the characteristic Raman peak at 1382 cm-1. This modified substrate has a low detection limit (0.2 ng cm-2 of thiram for apple and tomato), outstanding uniformity (relative standard deviation = 8.9%) and thermal stability (relative standard deviation = 0.9%). Graphical abstract Schematic representation of using a aluminum foil modified with ZnO nanosheets as a flexible and recyclable substrate for SERS analysis of pollutants. The substrate can be cleaned after use by UV irradiation.

Comparison of different methods of DNA recovery and PCR amplification in STR profiling of casings-a retrospective study.

Casings represent common evidence in a forensic laboratory, due to high frequency of firearms usage during perpetration of criminal offenses. Possible DNA evidence from casings is compromised by degradation, inhibition, and initial low-quantity deposition of biological material. For that reason, in the last 15 years, scientists have been trying to optimize procedures for recovery and amplification of DNA possibly present on its surface. In this study, we share our 12-year experience done on a total of 698 casework casings, comparing two DNA recovery methods commonly used-soaking and swabbing, as well as efficacy of two commercially available DNA amplification kits (AmpFLSTR® Identifiler® and AmpFLSTR® Identifiler® Plus kits). Of all analyzed casings, 30 were excluded as 28 (4%) matched the victims' DNA profiles and 2 (0.3%) samples were proved to be contaminated by technicians. Overall success in obtaining interpretable DNA profiles was 15.6% (104/668) (13.8% (55/399) for AmpFLSTR® Identifiler® Plus combined with soaking, 22% (33/150) for AmpFLSTR® Identifiler® Plus combined with swabbing, and 13.4% (16/119) using AmpFLSTR® Identifiler® kit and swabbing recovery method). Our data suggest the importance of both DNA recovery methods and amplification kits used, and point out swabbing of casings combined with AmpFLSTR® Identifiler® Plus kit as methods of choice. Nonetheless, our results are based on real casework and are prone to uncontrolled variables.

Validated measurements of microbial loads on environmental surfaces in intensive care units before and after disinfecting cleaning.

AIMS: Preanalytic aspects can make results of hygiene studies difficult to compare. Efficacy of surface disinfection was assessed with an evaluated swabbing procedure. METHODS AND RESULTS: A validated microbial screening of surfaces was performed in the patients' environment and from hands of healthcare workers on two intensive care units (ICUs) prior to and after a standardized disinfection procedure. From a pure culture, the recovery rate of the swabs for Staphylococcus aureus was 35%-64% and dropped to 0%-22% from a mixed culture with 10-times more Staphylococcus epidermidis than S. aureus. Microbial surface loads 30 min before and after the cleaning procedures were indistinguishable. CONCLUSIONS: The quality-ensured screening procedure proved that adequate hygiene procedures are associated with a low overall colonization of surfaces and skin of healthcare workers. Unchanged microbial loads before and after surface disinfection demonstrated the low additional impact of this procedure in the endemic situation when the pathogen load prior to surface disinfection is already low. SIGNIFICANCE AND IMPACT OF THE STUDY: Based on a validated screening system ensuring the interpretability and reliability of the results, the study confirms the efficiency of combined hand and surface hygiene procedures to guarantee low rates of bacterial colonization.

Evaluation of self-swabbing coupled with a telephone health helpline as an adjunct tool for surveillance of influenza viruses in Ontario.

BACKGROUND: Calls to a telephone health helpline (THHL) have been previously evaluated for the ability to monitor specific syndromes, such as fever and influenza-like-illness or gastrointestinal illness. This method of surveillance has been shown to be highly correlated with traditional surveillance methods, and to have potential for early detection of community-based illness. Self-sampling, or having a person take his/her own nasal swab, has also proven successful as a useful method for obtaining a specimen, which may be used for respiratory virus detection. METHODS: This study describes a self-swabbing surveillance system mediated by a nurse-led THHL in Ontario whereby syndromic surveillance concepts are used to recruit and monitor participants with influenza-like illness. Once recruited, participants collect a nasal specimen obtained by self-swabbing and submit for testing and laboratory confirmation. Enumeration of weekly case counts was used to evaluate the timeliness of the self-swabbing surveillance system through comparison to other respiratory virus and influenza surveillance systems in Ontario. The operational efficiency of the system was also evaluated. RESULTS: The mean and median number of days between the day that a participant called the THHL, to the day a package was received at the laboratory for testing were approximately 10.4 and 8.6 days, respectively. The time between self-swab collection and package reception was 4.9 days on average, with a median of 4 days. The self-swabbing surveillance system adequately captured the 2014 influenza B season in a timely manner when compared to other Ontario-based sources of influenza surveillance data from the same year; however, the emergence of influenza B was not detected any earlier than with these other surveillance systems. Influenza A surveillance was also evaluated. Using the THHL self-swabbing system, a peak in the number of cases for influenza A was observed approximately one week after or during the same week as that reported by the other surveillance systems. CONCLUSION: This one-year pilot study suggests that the THHL self-swabbing surveillance system has significant potential as an adjunct tool for the surveillance of influenza viruses in Ontario. Recommendations for improving system efficacy are discussed.

Direct Analysis in Real Time (DART) of an Organothiophosphate at Ultrahigh Resolution by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry and Tandem Mass Spectrometry.

Direct analysis in real time (DART) is a recently developed ambient ionization technique for mass spectrometry to enable rapid and sensitive analyses with little or no sample preparation. After swab-based field sampling, the organothiophosphate malathion was analyzed using DART-Fourier transform ion cyclotron resonance (FT-ICR) mass spectrometry (MS) and tandem mass spectrometry (MS/MS). Mass resolution was documented to be over 800,000 in full-scan MS mode and over 1,000,000 for an MS/MS product ion produced by collision-induced dissociation of the protonated analyte. Mass measurement accuracy below 1 ppm was obtained for all DART-generated ions that belonged to the test compound in the mass spectra acquired using only external mass calibration. This high mass measurement accuracy, achievable at present only through FTMS, was required for unequivocal identification of the corresponding molecular formulae.