Aerosol Contamination Research Articles

Performance of the Flash10 COVID-19 point-of-care molecular test

After the COVID-19 pandemic, fever clinics urgently require rapid nucleic acid tests to enhance their capacity for timely pathogen detection. This study evaluated the analytical performance and clinical utility of the Flash10 SARS-CoV-2 point-of-care test (Flash10 POCT) for detecting SARS-CoV-2 in patients with fever in the adult fever clinic in Beijing Tsinghua Changgung Hospital from August 1 to August 30, 2023. The analytical performance and clinical utility of the Flash10 POCT for detecting SARS-CoV-2 were assessed in 125 patients with fever syndrome in the adult fever clinic. The Flash10 POCT demonstrated an analytical precision of 3.1% for the Ct values of the ORF1ab gene and 2.9% for the Ct values of the N gene in SARS-CoV-2 nucleic acid testing. Furthermore, the Flash10 POCT demonstrated a lower limit of detection (LoD) of 100 copies/mL, with no detected aerosol contamination leakage. Of the 125 patients (median age 61.9 years, 52% male and 48% female), both the Flash10 POCT and RT-PCR tests yielded positive results for 100 patients and negative results for 25 patients (Fisher’s exact test, p < 0.0001). The median turn-around-time for the Flash10 POCT was significantly shorter, at 1.05 h, compared to 16.15 h required for RT-PCR tests (Wilcoxon signed rank test, p < 0.0001). The Flash10 POCT showed high analytical performance, achieving a 100% detection rate for SARS-CoV-2 compared to RT-PCR tests, while also exhibiting a significantly shorter turn-around-time. Implementing the Flash10 POCT had the potential to expedite the care of adults presenting with fever.

Benchtop to at-home test: Amplicon-depleted CRISPR-regulated loop mediated amplification at skin-temperature for viral load monitoring

CoRPLA (CRISPR-regulated One-pot Recombinase Polymerase Loop-mediated Amplification) is an amplicon-depleted skin-temperature operated iNAAT designed for at-home testing. It uses specially designed loop primers to enhance isothermal amplification, triggering Cas12 for in-situ amplicon depletion and signal amplification. This method addresses issues like amplicon-derived aerosol contamination and complex assay formats, enabling quantitative detection with sub-attomolar sensitivity (0.5 cps/μL). CoRPLA employs a DNA hydrogel wearable tape for real-time, colorimetric readout, allowing visual differentiation of pathogen loads. It was validated with clinical samples for SARS-CoV-2, RSV, influenza A, and HPV, successfully identifying multi-level viral loads of the positive cases with results consistent with qPCR. Offering high sensitivity while eliminating false positives from aerosol contamination, CoRPLA bridges the molecular assay from benchtop to home for daily viral infections monitoring.

Prospective Study of Contamination Spread Produced During Open Surgical vs Conventional Percutaneous vs Novel Percutaneous Tracheostomy During COVID-19 Era in a Pig Model.

Because of concerns about provider contamination during tracheostomy procedures in a pandemic such as COVID-19, it is essential to objectively evaluate aerosol generation in all tracheostomy approaches, including newly developed tracheostomy procedures. We performed open surgical tracheostomy (OST), conventional percutaneous tracheostomy (CPT), and novel percutaneous tracheostomy (NPT), a modification of CPT designed to reduce contamination spread, in pig models and then compared the degree of contamination to providers using Glo Germ (Glo Germ, Moab, UT, USA). Six Yorkshire pigs were used for data collection. Either OST, CPT or NPT was performed on the pigs by the same team including a surgeon, anesthesiologist, and respiratory therapist. A mixture of Glo Germ and water was administered via atomizer into the oral cavity to the tracheal bifurcation before each procedure, and additionally dispersed via an aersolizer in the trachea and lungs through the endotracheal tube before and throughout the procedure. Before and immediately after each procedure, pre-specified body parts of the providers were photographed and two independent examiners blindly evaluated the photographs to determine degree of Glo Germ contamination using a 3-point Likert scale. Total contamination among provider team average score (min. 0, max. 2), was significantly lower for OST than CPT (0.29 ± 0.59 vs 0.63 ± 0.65; P<0.01) or NPT (0.29 ± 0.59 vs 0.59 ± 0.66; p <0.01). No significant difference was seen in overall contamination of any provider between CPT and NPT (0.63 ± 0.65 vs 0.59 ± 0.66; p =0.92). Our results suggest that OST causes significantly less aerosol contamination to providers than either CPT or NPT.

Risk perceptions and preventive preparedness toward anthrax re-emergence within the lens of one health in Northcentral Nigeria.

Anthrax has re-emerged in domestic ruminants in Nigeria with public health concerns. This study assessed stakeholders' perceptions and preventive preparedness toward further resurgence and spread at the human-animal-environment interface. A questionnaire-based cross-sectional study was conducted in Nigeria. Descriptive and analytical statistical analyses were performed at 95% confidence levels. All the 384 recruited stakeholders responded. The majority (96.2%) of animal health practitioners (AHPs) and 56.7% of pastoralists were aware of the recent anthrax re-emergence in Nigeria (p < 0.001). Noteworthy, 88.5% of the AHPs and 32.2% of pastoralists mentioned that anthrax has an environmental component in its transmission to humans and animals. From the environmental perspective, 87.7% of AHPs and 24.0% of pastoralists significantly perceived that soil and aerosol contamination with anthrax spores are highly plausible explanation routes for its re-emergence. Extreme weather events (high rainfall, flooding, winds, and drought) (p = 0.001); grazing of livestock on pastures grown on contaminated soil (p < 0.001), transboundary movement and trade of animals (p = 0.001); introduction of new animals into the herds without quarantine (p = 0.001); and bioterrorism (p < 0.001) were more likely to influence the re-emergence and spread of anthrax. To tackle gaps in knowledge and risk perceptions, and address the socio-economic and anthropogenic drivers, cooperation and collaborations through the lens of the One Health approach are needed. The partnership will promote an integrated disease surveillance system from planning to implementation for the realization of elimination or reduction of the burden of anthrax and other zoonoses in Nigeria and contribute to achieving food safety, food security, and public and ecosystem health.

Bacterial and fungal aerosols in poultry houses: PM2.5 metagenomics via single-molecule real-time sequencing

Microbial aerosol contamination is a common problem in poultry farms, posing potential health risks to poultry and their caretakers. Exploring the distribution and diversity of the microbial community in poultry farm aerosols is crucial for effective mitigation strategies. The composition of bacterial and fungal aerosols is poorly understood, particularly the prevalence of potential pathogenic microorganisms in fine particulate matter (PM2.5) in various types of poultry houses. In this study, 27 PM2.5 samples were collected from 5 chicken houses and 4 duck houses in Shandong Province, China. Species-level diversity of bacterial and fungal components in PM2.5 samples was determined using advanced single-molecule real-time sequencing (SMRT) technology, based on the 16S and internal transcribed spacer 1 (ITS) ribosomal genes. Microbial diversity and community composition varied significantly across the different poultry house. Notably, duck houses had higher concentrations (p < 0.01) of PM2.5 (92.8-143.1 μg/m3) than chicken houses (42.0-56.4 μg/m3). Furthermore, microbial variation in PM2.5 samples was associated with the type of poultry facility. The predominant pathogenic microorganisms included Aspergillus sydowii, Penicillium sp., Aspergillus insolitus, Cladosporium sp., Aspergillus sp., Aspergillus pseudoglaucus, Cladosporium sp. C4092-2-PD1, and Colletotrichum sp., all of which were classified as second category of pathogens. Aspergillus sydowii and Penicillium sp. were the dominant species in chicken houses, while Cladosporium sp., Aspergillus sp., and Aspergillus pseudoglaucus were the dominant species identified in duck houses. To our knowledge, this study is the first to investigate bacterial and fungal diversity in PM2.5 samples collected from various types of poultry houses. These findings advance our understanding of poultry environmental microbiology and have important implications for safeguarding the health of both poultry and their caretakers.

Experimental comparison of aerosol transmission in displacement ventilation and mixing ventilation in a meeting scenario

The performance of displacement ventilation (DV) and mixing ventilation (MV) in aerosol contamination control was compared. The considered contamination was an aerosol emitted from a person in meeting scenarios of two and four persons. The experiments were carried out in a full-scale room measuring 4.4 m × 5.2 m × 2.9 m. Particle counting was carried out at 41 measurement points in the room to assess the concentration field of particles in the room and in the inhalation zone. The influence of the supply airflow rate and the number of activated thermal mannequins on the contaminant removal effectiveness were examined. The main conclusion is that the DV system was superior to the MV system in reducing contamination. An enhanced contaminant removal effectiveness of displacement ventilation was observed for all supply air flow rates and increased with higher airflow rates. At lower airflow rates, contamination in the inhalation zone was reduced by 40% compared to mixing ventilation, while at higher flow rates, the reduction surpassed 50%. This study highlights the advantages of displacement ventilation in terms of contamination control, infection prevention and energy efficiency, emphasising the importance of ventilation system selection for indoor environments, particularly those where airborne transmission risks are prevalent.

Prevention of contaminated aerosol and the transmission during nebulized therapy in hospital settings: a systematic review

Inhalation nebulization therapy is important for administering medications to patients in aerosolized form. However, there are persistent apprehensions in healthcare settings regarding aerosol contamination because of the significant infection risk. Despite rigorous adherence to established hospital protocols, concerns about potential contamination and transmission persist, raising considerable apprehension about nosocomial pneumonia. This condition shows the urgent need for implementing highly effective strategies to ensure patient safety during nebulization therapy. Therefore, this study aimed to review current investigations, focusing on interventions to mitigate aerosol contamination and minimize the transmission of contaminated aerosols.Adhering to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines, this systematic review included an exhaustive analysis of randomized and non-randomized clinical trials as well as, simulated experimental and in vitro studies published in English in the past decade. A meticulous search was conducted across four major databases, namely ScienceDirect, Cumulative Index to Nursing &amp; Allied Health (CINAHL), PubMed, and Scopus. A total of 37 pertinent studies were identified and subjected to rigorous analysis.The preventive measures include a range of strategies, such as the use of masks by therapists, thorough disinfection of nebulizers, integration of filters, and regular environmental cleaning in the vicinity of the patient.In conclusion, these multifaceted interventions are significant in preventing the administration of contaminated aerosols and curbing the proliferation of infectious agents in the hospital environment.

Enhanced Two-Step LAMP-CRISPR Assay with an Engineered Zst Polymerase for Contamination-Free and Ultrasensitive DNA Detection.

Current loop-mediated isothermal amplification (LAMP)-coupled clustered regularly interspaced short palindromic repeats (LAMP-CRISPR) biosensing in two-step or one-step formats has been applied to next-generation accurate molecular diagnosis. However, two-step LAMP-CRISPR assays intrinsically confront aerosol contamination, while one-step assays possess a compromised detection performance. To this end, we propose an enhanced two-step LAMP-CRISPR assay (ETL-CRISPR) with an engineered Zst polymerase to mediate ultrasensitive DNA detection and thoroughly eliminate aerosol contamination. Instead of supplementing any dTTP, the newly engineered Zst polymerase can efficiently polymerize four oligonucleotides (dATP, dCTP, dGTP, and dUTP), thereby enabling contamination-free and ultrasensitive ETL-CRISPR assay. By targeting the L1 gene of human papillomaviruses (HPV) 16 and the E7 gene of HPV18, our ETL-CRISPR assay achieves high specificity and single-copy level sensitivity within 1 h. Furthermore, we validated the assay by using 85 HPV clinical swab samples with an accuracy of 98.8%, which is comparable to the real-time quantitative polymerase chain reaction. Therefore, ETL-CRISPR provides a straightforward strategy for the contamination-free and ultrasensitive point-of-care diagnosis of clinical pathogens.

A novel system with robust compatibility and stability for detecting Sugarcane yellow leaf virus based on CRISPR-Cas12a.

Sugarcane yellow leaf virus (SCYLV) can reduce sugarcane productivity. A novel detection system based on reverse transcription-multienzyme isothermal rapid amplification (RT-MIRA) combined with CRISPR-Cas12a, named RT-MIRA-CRISPR-Cas12a, was developed. This innovative approach employs crude leaf extract directly as the reaction template, streamlining the extraction process for simplicity and speed. Combining RT-MIRA and CRISPR-Cas12a in one reaction tube increases the ease of operation while reducing the risk of aerosol contamination. In addition, it exhibits sensitivity equivalent to qPCR, boasting a lower detection limit of 25 copies. Remarkably, the entire process, from sample extraction to reaction completion, requires only 52-57 minutes, just a thermostat water bath. The result can be observed and judged by the naked eye.IMPORTANCESugarcane yellow leaf disease (SCYLD) is an important viral disease that affects sugarcane yield. There is an urgent need for rapid, sensitive, and stable detection methods. The reverse transcription-multienzyme isothermal rapid amplification combined with CRISPR-Cas12a (RT-MIRA-CRISPR-Cas12a) method established in this study has good specificity and high sensitivity. In addition, the system showed good compatibility and stability with the crude leaf extract, as shown by the fact that the crude extract of the positive sample could still be stably detected after 1 week when placed at 4°C. RT-MIRA-CRISPR-Cas12a, reverse transcription polymerase chain reaction (RT-PCR), and reverse transcription-quantitative polymerase chain reaction (RT-qPCR) were used to detect SCYLV on 33 sugarcane leaf samples collected from the field, and it was found that the three methods reached consistent conclusions. This Cas12a-based detection method proves highly suitable for the rapid on-site detection of the SCYLV.

Mobile lab: A novel pathogen assay using the nucleic acid automatic assay system assisted by a self-contained microfluidic cassette and chitosan decorating magnetic particles

Microfluidic pathogen detection cartridges with integrated switching valves have gained significant attention. Traditionally, these microfluidic cartridges utilized magnetic beads for a “bind-wash-elute-reaction” protocol to extract DNA but often lacked PCR-based aerosol contamination validation and focused primarily on isothermal applications. There is room to improve these systems, such as using novel DNA extraction methods and integrating PCR as the gold standard. In this study, we developed chitosan-decorated magnetic (CDM) particles and incorporated them into a mass-produced, self-contained microfluidic cartridge for the first time. Our results demonstrated that the DNA can be efficiently enriched from the lysis buffer (pH 5) using 75 % degree of deacetylation and 1 % of chitosan-modified CDM particles (20 μL). The DNA extraction efficiency exceeded 90 %. The CDM particles enabled “in-situ PCR”, simplifying the on-chip detection process to a “bind-wash-reaction” protocol. Additionally, we developed a nucleic acid automatic assay system (NA3 system) featuring a fluid thermocycler, four-channel optical detection, magnet, microcontroller, and software. The entire assay, from sample preparation to detection, can be completed within approximately 60 minutes, simultaneously detecting 16 targets. The performance of the NA3 system was better than that of the pristine magnetic microparticles-based assay and the commercial magnetic beads kit-based assay. The linearity, specificity, multi-pathogen detection, and user-friendliness of the NA3 system were tested. We validated the NA3 system's performance in detecting food-borne and serum infectious pathogens, demonstrating its potential for use in resource-limited settings.

LAMP assay coupled CRISPR/LbCas12a system in a single-tube method for visual detection of meat adulteration

Meat adulteration frequently infringes upon consumers' rights and interests, disrupts the market, and violates religious beliefs. However, implementing on-site detection of meat samples remains technically challenging for widespread applications. Here, we developed a point-of-care testing (POCT) platform for fast, highly sensitive, and on-site detection of meat adulteration, based on loop-mediated isothermal amplification (LAMP) coupled the clustered regularly interspaced short palindromic repeats (CRISPR) system in a single-tube. This platform integrates the efficient exponential amplification capacity of LAMP and the high specificity recognition capability of CRISPR, reducing the false positive results and minimizing LAMP aerosol contamination risk. Pork was selected as the target for adulteration detection, qualitative detection by the naked eye can be achieved within 1 h from sample collection to results with a sensitivity as high as 0.05 % to detect binary mixtures of pork in beef and the limit of DNA detection was at 50 pg/μL which 10 times higher than the individual LAMP assay. This work is expected to offer a powerful universal POCT platform for addressing a range of meat adulteration issues, suitable for enhancing food safety measures.

Detection of foodborne methicillin-resistant Staphylococcus aureus via fluorescence-encoded microsphere and Argonaute-mediated decoding

Molecular diagnosis of foodborne methicillin-resistant Staphylococcus aureus (MRSA) is crucial for controlling its dissemination and ensuring food safety. However, existing genetic methods are limited by susceptibility to aerosol contamination and restricted to single-gene detection. Herein, a fluorescent biosensor employing fluorescence-encoded microspheres and Argonaute-mediated decoding is developed, enabling ultrasensitive, accurate, and duplex detection of MRSA genes. This assay utilizes a target-triggered polymerization/nicking reaction to cyclically produce specific guide DNA, guiding Argonaute protein to site-specifically cleave the molecular beacon on the microsphere, thereby decoding a fluorescent signal. Notably, the fluorescence-encoded microsphere, designed via on-tetrahedron rolling circle amplification, achieves high fluorescence loadings in a unit area. This biosensor demonstrates simultaneous detection of two unamplified MRSA genes, mecA and femA, at concentrations as low as 0.63 fM and 0.48 fM, respectively. Moreover, the method exhibited excellent recoveries in milk, egg, and pork samples ranging from 73% to 112%, highlighting its practicability in real scenarios.

A rapid simultaneous detection of duck hepatitis A virus 3 and novel duck reovirus based on RPA CRISPR Cas12a/Cas13a

The mixed infection of duck hepatitis A virus 3 (DHAV-3) and novel duck reovirus (NDRV) has caused significant losses to the global duck farming industry. On-site point-of-care testing of viruses plays a crucial role in the early diagnosis, prevention, and disease control. Here, we proposed an RPA-CRISPR Cas12a/Cas13a one-pot strategy (DRCFS) for rapid and simultaneous detection of DHAV-3 and NDRV. This method integrated the reaction of RPA and CRISPR Cas12a/Cas13a in a single tube, eliminating the need to open the lid during the intermediate processes and thereby avoiding aerosol contamination. On this basis, we proposed a dual RPA-CRISPR strategy coupled with a lateral flow analysis platform (DRC-LFA). This circumvented the necessity for complex instruments, enabling direct visual interpretation of results, making the test more accessible and user-friendly. Our findings demonstrated that the DRCFS method could detect DHAV-3 and NDRV at concentrations as low as 100 copy/μL, while DRC-LFA achieved limit of 101 copies/μL within 35 min. Furthermore, when DRCFS, DRC-LFA, and qPCR were employed collectively for clinical samples analysis, all three methods yielded consistent results. The specificity, sensitivity, and user-friendly of these methods rendered them invaluable for on-site virus detection.

Rapid Discriminative Identification of the Two Predominant Echinococcus Species from Canine Fecal Samples in the Tibetan Region of China by Loop-Mediated Isothermal Amplification-Lateral Flow Dipstick Assay.

Echinococcosis poses a significant concern in the fields of public health and veterinary care as it can be transmitted between animals and humans. The primary endemic subtypes are cystic echinococcosis (CE) and alveolar echinococcosis (AE), which result from infestation by Echinococcus granulosus and Echinococcus multilocularis, respectively. A prominent epidemic of echinococcosis greatly affects the Tibet Autonomous Region (TAR) in China. A new technique called the loop-mediated isothermal amplification-lateral flow dipstick (LAMP-LFD) test is introduced in this research to differentiate between E. granulosus and E. multilocularis using their repetitive genetic sequences. The test is characterized by its portable nature, simple operation, quick result production, high sensitivity, and low susceptibility to aerosol contamination. The LAMP-LFD method demonstrated an exceptional minimal detection limit, reaching levels as low as approximately 1 fg/μL (femtogram per microliter) of genomic DNA. The assay's specificity was assessed, and no cross-reactivity was seen. A total of 982 dog fecal samples were collected from 54 counties in the TAR region between July 2021 and June 2022. The established method underwent validation using a commercially available ELISA kit. The agreement rate between the LAMP-LFD and ELISA methods was 97.25%, with a sensitivity of 96.05% and a specificity of 97.35%. The assay described in this study improves specificity by using a double-labeled probe, and it reduces the risk of false-positive results caused by aerosol contamination through the use of a sealed device. This makes it a suitable choice for quickly and accurately identifying the two main types of Echinococcus in field settings.

Aerosol interference with open-path eddy covariance measurement in a lake environment

Open-path eddy covariance (OPEC) is widely used for measuring trace gas fluxes between the surface and the atmosphere. At a lake in Eastern China (Lake Taihu), the CO2 flux measured with OPEC was often negative at night (with values as low as −22.1 μmol m−2 s−1) and was coherent across the whole lake, as if the lake were a large sink of atmospheric CO2. The purpose of this study is to investigate the cause of this negative flux phenomenon. In addition to OPEC, we also used closed-path eddy covariance (CPEC) and the transfer coefficient (TC) method to measure the flux. The results show that the persistent negative CO2 flux phenomenon was observed with OPEC but not with CPEC or TC. Because air drawn into the CPEC analyzer was filtered but the OPEC analyzer was influenced by aerosol contamination, the most logical explanation was that particles deposited on the optical lens of the OPEC analyzer changed its cross-sensitivity to water vapor. The direct evidence of this interference was a strong positive correlation between the OPEC analyzer's signal strength and the CO2 mixing ratio observed at 10 Hz. We suggest that it is possible to perform post-field correction to this negative flux bias using the 10 Hz signal strength data. In comparison, an OPEC system at a nearby land site did not experience aerosol interference due to low water vapor flux at night and lack of hygroscopic growth of particles on the optical lens in low humidity conditions. The type of aerosol interference reported here may also occur in high humidity and high pollution conditions elsewhere.

Revolutionizing respiratory health research: “commercially-available lung-on-a-chip and air-liquid interface systems”

Inhalation of ultrafine particles, aerosol contaminants, and cigarette smoke can induce respiratory diseases. As humans are constantly exposed to various exogenous substances, it is crucial to study their impacts on respiratory diseases and airway dysfunction. Recently, organ-on-a-chip technology has been applied in many research studies to understand disease mechanisms, drug screening, and drug testing. The combination of organ-on-a-chip technology and the air-liquid interface (ALI) culture method is emerging as a new platform for realistically mimicking the microenvironment and physiological motions of the human lungs. Breathing motion can be simulated through cyclic stretching, while blood flow can be replicated using channel flow within the chip. The ALI system is critical for mucociliary differentiation, pseudostratified morphology, and epithelial barrier function development. The combination of organ-on-a-chip technology and ALI systems allows the integration of cyclic stretch as a breathing motion and microfluidic channels as circulatory systems. The chip system can also integrate the lung epithelial cells, extracellular matrix, and microstructures, providing microenvironments such as fibroblast, collagen, and immune cells to the epithelial cells. This review discusses chip systems as effective tools for recapitulating human lung environments and how they are applied in biological studies against various pulmonary diseases such as infections or inflammation, fibrosis, and malignancy.

DNA Polymerase-Endonuclease Efficiently Synthesizes DNA to Prepare DNA Materials and Develop Novel Signal Amplification System.

DNA biosynthesis, a focus of fundamental and applied research, typically involves DNA polymerases by using templates, primers, and dNTPs. Some polymerases can polymerize dNTPs for DNA de novo synthesis, although this is generally to occur randomly. This novel synthesis method has garnered our attention and practical use. Herein, we observed that the addition of endonuclease significantly enhances the efficiency of the de novo synthesis reaction catalyzed by the DNA polymerase. We further investigated the reaction conditions that influence this efficiency. Building on the optimal reaction conditions, we developed a rapid and efficient strategy for preparing DNA hydrogel. Further, coupled with the CRISPR-Cas system, we developed a nucleic acid signal amplification system characterized by versatility, sensitivity, specificity, and no risk of aerosol contamination. We successfully detected viral nucleic acids in clinical samples. In summary, our study demonstrates the significant potential of DNA polymerase- and endonuclease-catalyzed DNA de novo synthesis in diverse applications.

Quantitative evaluation of the impact of indoor relative humidity on deposition of aerosols generated during tooth grinding in a real-world clinical setting.

Exposure to aerosol particles generated from tooth grinding has a negative impact on the health of dental personnel. The aim of this study was to quantitatively analyze the impact of indoor relative humidity (IRH) on the deposition of these suspended particles in a well-controlled dental environment. In this study, a humidity control system was employed to effectively regulate and maintain indoor relative humidity (IRH). A novel computer-assisted numerical control system was developed to pre-treat the molar specimens, and accurately simulate clinical tooth grinding procedures. Each procedure was performed in triplicate, with an online real-time particle counter (ORPC; TR-8301, TongrenCo.) measuring aerosol production. All testing devices were controlled remotely. The data obtained were statistically analyzed using descriptive statistics and non-parametric tests (Kruskal-Wallis/ Dunn's post hoc test with Bonferroni correction, p < 0.05). The findings showed that with increasing IRH, the maximum peak concentration of aerosol particles decreased by 397% from 6.51 × 107 particles/m3 at 30% to 1.64 × 107 particles/m3 at 80%. The Kruskal-Wallis test results indicated a statistically significant effect of IRH on the aerosol increment (p < 0.05). Increasing the IRH level can effectively promote the deposition of aerosol particles, with a return to baseline within 15min after reaching 60% or above. Our study suggested that maintaining IRH above 70% during the cleaning process, allowing natural recovery to ambient humidity levels within 15min after cleaning, and taking basic precautions, may lead to an adequate reduction in the possible health risks of aerosol contamination.

Detection accuracy and clinical applications of DP-TOF mass spectrometry.

Matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) is currently used in clinical microbiology laboratories. This study aimed to determine whether dual-polarity time-of-flight mass spectrometry (DP-TOF MS) could be applied to clinical nucleotide detection. This prospective study included 40 healthy individuals and 110 patients diagnosed with cardiovascular diseases. We used DP-TOF MS and Sanger sequencing to evaluate 17 loci across 11 genes associated with cardiovascular drug responses. In addition, we used DP-TOF MS to test 998 retrospectively collected clinical DNA samples with known results. A, T, and G nucleotide detection by DP-TOF MS and Sanger sequencing revealed 100% concordance, whereas the C nucleotide concordance was 99.86%. Genotyping based on the results of the two methods showed 99.96% concordance. Regarding clinical applications, DP-TOF MS yielded a 99.91% concordance rate for known loci. The minimum detection limit for DNA was 0.4 ng; the inter-assay and intra-assay precision rates were both 100%. Anti-interference analysis showed that aerosol contamination greater than 1013 copies/µL in the laboratory environment could influence the results of DP-TOF MS. The DP-TOF MS platform displayed good detection performance, as demonstrated by its 99.96% concordance rate with Sanger sequencing. Thus, it may be applied to clinical nucleotide detection.

Visual identification for species and sex derived from bloodstain based on phosphate-mediated isothermal amplification colorimetric system

Species and sex confirmation of the biological specimen play a crucial role in crime investigation. However, the specimen found in the scene is always trace quantity, which is hard to be analyzed by current methods. Moreover, the time-consuming DNA extraction, sophisticated apparatus, and complex data processing make it difficult to satisfy the demand of speediness and convenience for point-of-care tests. In this study, we first exhibit a phosphate-based visual system for field-based species and sex identification derived from trace bloodstain. By introducing phosphate ion-based colorimetry into loop-mediated isothermal amplification (LAMP) for result interpretation, not only the bloodstain can be directly submitted to mitochondrial variant amplification owing to the enhanced amplification efficiency by pyrophosphate ion hydrolyzation, but also the colorimetric signal can be recognized by the naked eye for result output within 30 min through molybdophosphate generation. Aerosol contamination, the major conflict of LAMP, has been solved once and for all by integrating uracil-DNA glycosylase into this system that still holds on a constant temperature. As a demonstration, cytochrome b and Y-chromosomal amelogenin are employed to identify species and sex respectively, which has achieved a highly sensitive and specific distinguishability under a strong interferential background. Accurate results can be obtained from both the simulative degraded and dated specimen, which indicates that this novel system may serve as a promising tool in forensic practice.