2.1 Pathogens, cells and equipment
Staphylococcus albus was purchased from the Microbiology Institute of Guangdong (Guangdong, China). HAdV-5 was provided by the State Key Laboratory of Guangzhou (Guangzhou, China). Escherichia coli BJ5183 and Top 10 cells were purchased from Takara (Dalian, China). Cigarettes (Hongtashan, Yunnan, China) were purchased from a local retailer. Ad293 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) containing penicillin-G 100 U/ mL, streptomycin 100μg/mL and 10% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) and frozen using a BioFlash commercial freezing kit (Fibulas, New York, NY, USA). A Y09-301 laser dust particle counter and an Anderson six-level sampler were both purchased from Sujin (Jiangsu, China). Virus collection which specially collection the virus was purchased from Millipore (Millipore,MMAS,USA). Viruses were purified using standard CsCl isopycnic centrifugation.
2.2 Pediatric isolation bed
This new pediatric isolation bed was innervated by Guangzhou Angel biosafety Co.,Ltd. The bed is connected to a purification device that surrounds thechild’s head, exhaled vapors from the child are directly drawn into the purification device that captures the harmful microorganisms by a sterilization filtering apparatus. The purified air is then blown back into the ward, providing patients with clean air, as well as reducing cross infections by air transmission among patients in the same ward. The purification device of the bed can be operated under three different wind speeds to create a negative pressure at the air inlet port, with high speed 0.86/ms, medium speed (0.55 m/s), and low speed (0.35 m/s), and can also be shut off (0 m/s) as a control. Compared with air quality controlled at the whole room level, the isolation bed reduces energy consumption and has enormous potential for application.
2.3 Geometry model establishment
The relevant parameters in the geometric model, including the size of the bed, the size of the purification equipment, and the size and position of the air inlet hood , were established according to the known parameters of the actual product, and were reasonably simplified. The specific parameters are shown in Table 1.
The air supply position had little effect on the simulation results, and different air supply positions are not discussed in this simulation.
To study the influence of the geometric relationship between the child and the air inlet hood on the effect of the intaking airflow,the angle θ,was set at the child’s mouth between the longitudinal axis of the body and the highest point of the air inlet hood, according to the equipment and children's geometrical shape in this case.
Actually,the range of θ can be found to be approximately 44.8°–73.0°. Therefore, simulations and comparisons were performed by taking θ=50°, 60°, and 70°. The closer the child is to the air inlet hood, the larger θ is when the shape of the device is unchanged. The schematic θ and the meshing results of the model are shown in Figure 1 (taking θ=70° as an example). When θ was 50°, 60°, and 70°, the total number of body meshes was 3472824, 3341411, and 3487467, respectively. The mesh surrounding the human body was the densest, followed by the mesh around the bed, and the other mesh scatteringin the room were the most sparse.
2.4 Simulation condition settings
The effects of different relative positions and different purifying wind speeds on indoor wind speed, pressure and pollutant distribution were investigated in the simulation. The simulation conditions are shown in Table 2. Among them, according to the actual usage situation, the purifying air volume of the child’s bed was 280 m3/h, that is, the windspeed of the purifying equipment was 0.86 m/s, as represented in Case 6.
This simulation simplifies the patient’s breathing state, considering only the patient’s exhalation. The exogenous pathogenic microorganisms are represented by the tracer gas, N2O.
2.5 calculation equations
The governing equations are given in a vector form as follows:
See Formula 1 in the Supplemental Files
Where ϕ is a general scalar quantity, which can represent u,v,w,k,e,T and tracer gas concentration. The diffusion coefficients Γ_ϕ and the source term S_ϕ for each scalar are summarized in Table 3-1. The tracer gas with a 4% volume fraction was added in the exhaled air of the source manikin. The properties of the tracer gas in the simulation were set the same as N2O which was used in the experiment. Only steady state simulation was considered and the exhaled air was set at an average velocity of 0.86 m/s and a temperature of 25 ºC.
2.6 Validation experiment
A 30 m3 test chamber was selected to simulate the size of a general ward room, and the isolation effect of the device in actual usage by means of exposure to artificial pollutants. The airflow wind speed using the results of the previous simulations was set at 0.86 m/s and θ=70°. The isolation bed was placed into the test chamber with a cigarette smoke generator release port or a microbial aerosol generator (bacterial and viral pathogens) placed 20 cm above the middle of the isolation bed. The horizontal distances from the wall were greater than 1 m, and the vertical distance from the ground was about 140 cm. The test was repeated three times and the average value of the pass efficiency was calculated. All tests were performed at 25°C–26°C, and the doors, windows, and air conditioners were all closed/off during the test.
2.6.1 Cigarette smoke
Using the smoke of a Hongtashan cigarette as a source of pollution, the cigarette was ignited and the cigarette-smoke generating device was activated. The release port was placed in position A (Figure 2), 20 cm above the bed, at a similar height to the face of the patient. Simultaneously, the purification device of the isolation bed was switched on. After 1 hour, the air was sampled at points A, B, C, D, E, F, G and H (Figure 2.), using a dust particle counter with a flow rate of 2.83 L/min and a sampling time of 20s.
2.6.2 Bacteria
Bacteria were cultured for between four and seven generations at 37°C for 24 h as indicator bacteria. A bacterial suspension with concertration 1.0 *107 cfu/ml was prepared and added to the microbial aerosol generator, which was activated and positioned with the release port at position A (Figure 2). Simultaneously, the purification device of the isolation bed was switched on. After 1 hour, the sampling port was positioned at points A, B, C, D, E, F, G and H (Figure 2). An Anderson six-stage sampler was used to collect and determine the concentration of microorganisms in the room at a flow rate of 28.3 L/min for 3 min. The sample plates were placed in an incubator at 37°C for 24 h to count the colonies and calculate the concentration of microorganisms in the air.
2.6.3 Viruses
HAdV-5 was cultured in AD293 cells, purified and stored at −80°C. The 50% tissue culture infective dose of the virus was determined following the routine procedure, as previously described [17]. Purified HAdV-5 was dilution with PBS to prepare a mixed-pathogen suspension, which was placed in the activated microbial aerosol generator with the release port at position A (Figure 2). Simultaneously, the isolation bed was switched on. After 1 hour, air samples were taken with the sampling port positioned at A, B, C, D, E, F, G and H (Figure 2).A virus collection was used to collect virus. and determine the concentration of virus in the room at a flow rate of 100 L/min for 10 min. The number of viral genome copies at each time point was determined by real-time quantitative PCR (Q-PCR) using a universal adenovirus Q-PCR kit (Hexin Corporation, Guangzhou, China) on an Applied Biosystems 7500 real-time PCR system (Foster City, CA). Each assay was performed three times in duplicate. The quantification of the genome copy numbers of HAdV-5 was performed with HAdV type specific primers and purified HAdV genomic DNA as the standard curve.We obtain the template genomic DNA from Pubmed.
2.7 Statistical analysis
Statistical analysis was performed by t-test, using Prism 7 software (GraphPad, San Diego, CA, USA). P values (P) less than 0.05 were considered statistically significant.