• Biomedical Science Letters
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• v.22 no.3
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• pp.65-74
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• 2016

The critical aspects of biosafety and bio-containment have been increasingly important in recent years. Biological agents involved in biological research projects at the Nanyang Technological University (NTU) Singapore are usually those with low risks. Biosafety level 2 or BSL 2 laboratories are widely used. However, biosafety measures which refer to the implementation of laboratory practices and procedures, specific construction features of laboratory facilities and safety equipment must be in place to reduce the exposure of laboratory personnel, the public or the environment to potentially infectious agents or other biological hazards. It is also required to pay more attention to laboratory-acquired infections (LAIs) which may occur in research laboratories, clinical laboratories or animal facilities. BSL 2 audit and certification program is implemented as an internal exercise covering laboratories in the university where biological agents are handled or biological research works are carried out. We have put some efforts to raise biosafety standards university-wide in both laboratory infrastructure and laboratory practices to a higher level. Common audit findings are briefly discussed in this presentation.

• Journal of Preventive Medicine and Public Health
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• v.38 no.4
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• pp.449-456
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• 2005

Objectives : The biosafety level (BSL) practiced in microbiology laboratories in Korea according to the laboratory biosafety manual published by the World Health Organization (WHO) was evaluated using the data obtained by a survey. Methods : Under the advise of Clinical Laboratory Physicians, 144 types of microorganisms were screened based on the guidelines of biosafety in microbiological and biomedical laboratories published by the US Center for Disease Control and Prevention and classified into 1-4 risk groups. A questionnaire containing 21 questions in 5 areas was developed using the biosafety manual by published WHO. Of the 1,876 different organizations sent the survey, 563 responded to the survey (response rate: 30.0%). The species of microoganisms handled by as well as the biosafety level in microbiology laboratories were analyzed. Results : There were 123 species of microorganisms handled in microbiology labs in Korea. The BSL required in 512 microbiology labs was answered by the survey responders as the first grade in 33 labs (6.4%), 2nd in 437 (85.4%), 3rd in 42 (8.2%), and 4th in none. The average number of items satisfied was 12.2, showing only a 57.9% satisfactory rate and normal distribution. Conclusions : The state of overall observance of BSL in most microbiology labs of Korea was evaluated as lagging compared with the standard set up by WHO. Therefore, the Korean government need to produce and distribute a biosafety manual in microbiology laboratories and make efforts to prevent this threat through measures such as training in biosafety in microbiology labs.

• Korean Journal of Clinical Laboratory Science
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• v.50 no.2
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• pp.77-84
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• 2018

In Singapore, biosafety and biosecurity measures are controlled by the Biological Agents and Toxins Act (BATA) and other requirements by regulatory agencies. The law prohibits and otherwise regulates the possession, use, import, transhipment, transfer, and transportation of biological agents, inactivated biological agents, and toxins that are of public health concern. The law also defines the facility requirements for high risk biological agents and toxins. The containment facility (BSL 3) is a minimum requirement to handle biological agents that falls under Schedule 1 (Risk Group 3). The Nanyang Technological University School of Biological Sciences Biosafety Level 3 Facility (NTU-SBS BSL 3) was designed specifically for research involving potential hazardous biological materials. The facility requires yearly re-certification by an approved facility certifier to meet the local requirements and international biosafety standards for a containment facility in many instances. On the other hand, most NTU researchers conduct biological projects involving biological agents with low or moderate risk groups (Risk Groups 1 and 2 or biological agents described in schedule 3 and 4 of BATA) and GMOs, which need only a BSL 2 laboratory. BSL 2 laboratories are yet to be legally certified or registered in Singapore. Institutional Biosafety Committee (IBC) identifies the requirements; defines a minimum standard in the safe control of biological risks and registers all BSL 2 laboratories in the NTU. Therefore, under the guidance of the IBC, the University Biosafety and Biosecurity Programme includes the audit and certification program as a unique and an internal exercise to bring NTU biosafety to a higher level.

• Biomedical Science Letters
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• v.24 no.3
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• pp.270-274
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• 2018

Chlorine dioxide ($ClO_2$) gas is a neutral chlorine compound. $ClO_2$ gas was proven to effectively decontaminate different environments, such as hospital rooms, ambulances, biosafety level 3 laboratories, and cafeterias. In this study, to evaluate the effects of $ClO_2$ gas, bacteria of clinical importance were applied. Staphylococci, Streptococci and Bacillus strains were applied and Klebsiella, and others e.g., Escherichia coli, Shigella, Salmonella, Serratia were also done for the inhibitory analysis. Bacteria plates were applied with a hygiene stick, namely, "FarmeTok (Medistick/Puristic)" to produce $ClO_2$. $ClO_2$-releasing hygiene stick showed the very strong inhibition of bacterial growth but had different inhibitions to the bacteria above 96.7% except for MRSA of 90% inhibition. It is difficult to explain why the MRSA were not inhibited less than others at this point. It can be only suggested that more releasing $ClO_2$ should be essential to kill or inhibit the MRSA. B. subtilis, S. agalactiae, S. pyogenes, E. coli O157:H7, S. typhi (S. enterica serotype typhi) and S. marcesence were inhibited over 99%. This study will provide fundamental data to research growth inhibition by $ClO_2$ gas with bacteria of clinical importance value.

• Biomedical Science Letters
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• v.23 no.2
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• pp.96-103
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• 2017

Nocardia species (spp.) are opportunistic pathogen in immunocompromised hosts. The genus Nocardia contains more than 70 species. Nocardia takedensis has been recently reported as a new species of the genus Nocardia. In this study, we describes the first clinical isolate of N. takedensis from the skin lesion in Busan, Korea. For the identification of clinical isolate to the species level as N. takedensis, classical methods (colony morphology, biochemical characteristics, and antimicrobial susceptibility), molecular method (16S rRNA gene sequencing), and MS (mass spectrometry) analysis were conducted. Clinical isolates grew slowly on the culture media (5% sheep blood agar and chocolate agar) under 5% $CO_2$ condition. Especially, carotene pigmentation was detected well on the media. Using mass spectrometry, Nocardia isolate was not identified to the species level. However, molecular method based on 16S rRNA sequencing confirmed the isolate as N. takedensis correctly. N. takedensis isolate was partial positive for acid-fast bacilli on the Ziehl-Neelsen method. And it was observed to be resistance to amoxicillin/clavulanic acid and ciprofloxacin. Our results provide useful information to develop optimal identification protocol of N. takedensis in clinical diagnostic laboratories.

• Biomedical Science Letters
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• v.26 no.3
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• pp.149-156
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• 2020

We developed a simple and rapid method for detecting vancomycin resistance genes, such as vanA and vanB, using loop-mediated isothermal amplification (LAMP). To identify not only vancomycin resistance genes, but also the genus Enterococcus, primers were designed for vanA, vanB, and 16S rRNA. Screening for vancomycin susceptibility in Enterococcus was performed using Etest (bioMérieux Inc). The results of the LAMP assay were compared to those of real-time RT-PCR. The optimal conditions for the LAMP assay were 65℃ for 60 min. The detection limits of the LAMP assay for vanA, and vanB were 2 × 10² copies/reaction. Compared to RT-PCR, the sensitivities and specificities of LAMP for 16S rRNA, vanA, and vanB were 100/100%, 100/100%, and 100/100%, respectively. The vanA genotype-vanB phenotype accounted for 57.5% (46/80) of the vancomycin-resistant Enterococci samples collected from 2016 to 2019. In conclusion, the LAMP assay developed in this study showed high sensitivity and specificity for vancomycin-resistant genes. Moreover, due to the simplicity and rapidity of the LAMP assay, its use can be very useful in clinical microbiology laboratories.

• Biomedical Science Letters
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• v.26 no.4
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• pp.275-287
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• 2020

Since its introduction in the forensic field, quantitative PCR (qPCR) has played an essential role in DNA analysis. Quality of DNA should be evaluated before short tandem repeat (STR) profiling to obtain reliable results and reduce unnecessary costs. To this end, various human DNA quantification kits have been developed. Among these kits, the PowerQunat® System was designed not only to determine the total amount of human DNA and human male DNA from a forensic evidence item, but also to offer data about degradation of DNA samples. However, a crucial limitation of the PowerQunat® System is its high cost. Therefore, to minimize the cost of DNA quantification, we evaluated kit performance using a reduced volume of reagents (1/2-volume) using DNA samples of varying types and concentrations. Our results demonstrated that the low-volume method has almost comparable performance to the manufacturer's method for human DNA quantification, human male DNA quantification, and DNA degradation index. Furthermore, using a reduced volume of regents, it is possible to run 2 times more reactions per kit. We expect the proposed low-volume method to cut costs in half for laboratories dealing with large numbers of DNA samples.

• Biomedical Science Letters
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• v.25 no.4
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• pp.381-389
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• 2019

The physiological function test is the only patient contact area in the field of clinical laboratory. We need to recruit and encourage the experts due to requiring the expertise and long time for examination. However, there is currently no rule for estimating optimal workforce in the field of physiological function tests. The purpose of this study is to establish the basis of the studies for mid- to long-long term job creation in the field of ultrasound by evaluating the number of appropriate tests and appropriate workforce. We calculated the quantitative and qualitative workload for the number of appropriate tests and appropriate workforce using online questionnaire. All statistical analyses were performed using SPSS version 18.0 (SPSS Inc., Chicago, IL, USA). A total of 216 respondents were 48 (22.2%) male and 168 (77.8%) female. A total of 157 laboratories were 62 (39.5%) for echocardiography, 91 (58.0%) for the transcranial Doppler (TCD) and 4 (5.7%) for the carotid ultrasound. The mean number of appropriate tests was 10 ± 2 in the echocardiography, 9 ± 2 in TCD and 11 ± 2 in the carotid ultrasound. In addition, the number of laboratories required to recruit employees for appropriate workforce was 19 in echocardiography, 18 in TCD, and 0 in carotid ultrasound. The number of hospital required to recruit workforce were 7 primary hospitals, 22 secondary hospitals, 7 third hospitals. This study can be used as an important data as the first study at present time when the data on the workforce status and work environment of the ultrasonic laboratories is insufficient. Based on the quantitative and qualitative workloads, the number of appropriate tests and appropriate workforce can support mid- to long-long term job creation in the field of ultrasound.

• Biomedical Science Letters
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• v.22 no.2
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• pp.29-36
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• 2016

Among the several agents causing carbapenem resistance of Acinetobacter baumannii, the most common cause is OXA-23 ${\beta}$-lactamase, which is known to hydrolyze carbapenem. To effectively control dissemination of carbapenem-resistant Acinetobacter baumannii (CRAB), development of both rapid and easy-to-use detection methods are required. The aim of this study is to develop a novel immunochromatographic assay (ICA) for rapid detection of OXA-23 ${\beta}$-lactamase. Of the seven monoclonal antibodies (mAbs) screened by ELISA, four mAbs (4G6, 4H6, 6G4, 9A4) exhibited high reactivity. Of these four specific antibodies, the combination of 6G4/4G6 showed the greatest reactivity and this combination of mAbs (6G4/4G6 mAbs) was used to develop the OXA-23 ${\beta}$-lactamase ICA. Of 102 A. baumannii isolates tested, the OXA-23 ${\beta}$-lactamase ICA results were consistent with PCR analysis except one false positive and one false negative isolate. The overall sensitivity and specificity were 98.36% and 97.56%, respectively. In conclusion, to the best of our knowledge, we have developed the first specific antibody set to detect OXA-23 ${\beta}$-lactamase using an ICA kit. This novel ICA can be used as a reliable and easy-to-use immunological assay for detection of OXA-23 ${\beta}$-lactamase producing CRAB in clinical laboratories.

• Biomedical Science Letters
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• v.26 no.2
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• pp.114-119
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• 2020

Of the various types of mice used for genome editing, conditional knock-out (cKO) mice serve as an important model for studying the function of genes. cKO mice can be produced using loxP knock-in (KI) mice in which loxP sequences (34 bp) are inserted on both sides of a specific region in the target gene. These mice can be used as KO mice that do not express a gene at a desired time or under a desired condition by cross-breeding with various Cre Tg mice. Genome editing has been recently made easy by the use of third-generation gene scissors, the CRISPR-Cas9 system. However, very few laboratories can produce mice for genome editing. Here we present a more efficient method for producing loxP KI mice. This method involves the use of an HDR vector as the target vector and ssODN as the donor DNA in order to induce homologous recombination for producing loxP KI mice. On injecting 20 ng/µL of ssODN, it was observed that the target exon was deleted or loxP was inserted on only one side. However, on injecting 10 ng/µL of the target HDR vector, the insertion of loxP was observed on both sides of the target region. In the first PCR, seven mice were identified to be loxP KI mice. The accuracy of their gene sequences was confirmed through Sanger sequencing. It is expected that the loxP KI mice produced in this study will serve as an important tool for identifying the function of the target gene.