Development of a bench-top extra-cleanroom for DNA amplification
Hirokazu TakahashiTakahiro SatohHiroko KanaharaYuji KubotaTamaki HiroseHiroyuki YamazakiKimiko YamamotoYoshiko OkamuraTaketo SuzukiToshiro Kobori
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Abstract:
Prevention of airborne contamination has become an important factor in biotechnology; however, conventional laminar-airflow cabinets (LAF-cabinets) are no longer sufficient as a countermeasure against nano-sized airborne contaminants in the laboratory. Here we present a bench-top extra-cleanroom classified as ISO-1 that can prevent contamination from airborne nanoparticles. This bench-top extra-cleanroom consists of a novel clean-zone-creating system that is equipped with nanofibrous, nonwoven filters. In addition, the cleanroom is also equipped with an ionizer to prevent plasticware from collecting dust by electrostatic charge attraction. This combination of features allows the cleanroom to prevent DNA contamination derived from airborne nanoparticles. Our extra-cleanroom with ionizer could be useful in various areas of biotechnology that are easily affected by airborne contaminants.Keywords:
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This chapter contains sections titled: Introduction Definition of a cleanroom Classes of cleanroom Occupancy states The meaning of the cleanroom classification US Federal Standard 209 E British Standard 5295 >209< Classification of airborne particles according to ISO 14644-1 Cleanliness testing within cleanrooms Classification of pharmaceutical clean room Different types of cleanrooms Cleanrooms and clean zones Working in clean zones Conclusions
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The existing standard for cleanrooms is based on an unoccupied state where there are no people or furniture inside the room. However, the distribution of airflow and particles in a cleanroom may differ significantly in an occupied state with the presence of people and furniture. This paper first validates a numerical model for simulating the air distribution in a locally concentrated cleanroom. Then the indoor airflow and the pollutant concentration in a locally concentrated bio-clean operation room (BCOR) are simulated at both occupied and unoccupied states by the validated model. The comparison of results shows the environment is very different between the two cases. Thus, human bodies and furniture should be properly considered when evaluating the environment in the locally concentrated cleanroom.
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Using a modified dispersal chamber, the authors have studied the protective efficacy of cleanroom clothing systems. Study results show that the state of a cleanroom clothing system-new or much used-influences the protection efficacy of the system. Suitable combinations of cleanroom underwear and cleanroom garments also improve the protection of the clean environment against airborne contaminants from people.
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A sampler that detects and counts viable particles in the air of cleanrooms in real-time was studied. It was found that when the sampler was used to monitor airborne particles dispersed from a number of materials used in cleanrooms, including garments, gloves, and skin, the number of viable particles dispersed from these materials was greater than anticipated. It was concluded that a substantial proportion of these viables were of a non-microbiological origin. When the sampler was used to monitor a non-unidirectional airflow cleanroom occupied by personnel wearing cleanroom garments, it was found that the airborne viable concentrations were unrealistically high and variable in comparison to microbe-carrying particles simultaneously measured with efficient microbial air samplers. These results confirmed previously reported ones obtained from a different real-time sampler. When the real-time sampler was used in a workstation within the same cleanroom, the recorded viables gave results that suggest that the sampler may provide an effective airborne monitoring method, but more investigations are required. LAY ABSTRACT: The airborne concentrations measured by a real-time microbial air sampler within an operational, non-unidirectional airflow cleanroom were found to be unrealistically high due to a substantial numbers of particles of non-microbiological origin. These particles, which resulted in false-positive microbial counts, were found to be associated with a number of materials used in cleanrooms. When the sampler was used within a cleanroom workstation, the counts appeared to be more realistic and suggest that this type of real-time airborne microbial counter may provide a useful monitoring method in such workstations, but further investigations are required.
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The contamination of platinum with nuclear fission products and its decontamination has been studied. Platinum plates were contaminated by soaking in a solution of fission products. The contamination increased with increase of the soaking time, and was saturated in about 3 hr. It was confirmed by the methods of autoradiography, electron microscopy and optical microscopy that the contamination was markedly affected by the surface condition of platinum. The results of the experiments of the decontamination are summarized as follows: (1) Decontamination with tap water: A sample soaked in the solution of fission products only for a short time was easy to decontaminate and the amount of the residual contamination reached equilibrium more rapidly than in the long-soaked sample. (2) Decontamination with acids: HCl, HNO3, H2SO4, H3PO4 and HF were tried. The effect of temperature on the decontamination was very significant. The decontamination with HCl of high temperature was more effective than with the other acids. (3) Decontamination with fused salts: The decontamination with fused KHSO4 was more effective than with the acids and other fused salts. Only one immersion of the contaminated platinum plate into fused KHSO4 could remove 99.4% of the contamination. Finally, it may be confirmed that the degree of the contamination of platinum with fission products is very low and it is easy to decontaminate it with the decontaminating agents such as HCl and KHSO4.
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Prevention of airborne contamination has become an important factor in biotechnology; however, conventional laminar-airflow cabinets (LAF-cabinets) are no longer sufficient as a countermeasure against nano-sized airborne contaminants in the laboratory. Here we present a bench-top extra-cleanroom classified as ISO-1 that can prevent contamination from airborne nanoparticles. This bench-top extra-cleanroom consists of a novel clean-zone-creating system that is equipped with nanofibrous, nonwoven filters. In addition, the cleanroom is also equipped with an ionizer to prevent plasticware from collecting dust by electrostatic charge attraction. This combination of features allows the cleanroom to prevent DNA contamination derived from airborne nanoparticles. Our extra-cleanroom with ionizer could be useful in various areas of biotechnology that are easily affected by airborne contaminants.
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