[논문]면봉시료에서 세균의 보존을 위한 최적 보관 온도와 채취 시약의 비교

이영주; 유희상; 이송희; 이소립; 이한; 성호중; 강희규; 현성희

doi:10.15324/kjcls.2021.53.4.326

면봉시료에서 세균의 보존을 위한 최적 보관 온도와 채취 시약의 비교
Comparison of Optimal Storage Temperature and Collection Reagents for Living Bacterial Cells in Swab Samples 원문보기

Korean journal of clinical laboratory science : KJCLS = 대한임상검사과학회지, v.53 no.4, 2021년, pp.326 - 332

이영주 (을지대학교대학원 임상병리학과) , 유희상 (을지대학교대학원 BK21 프로그램 시니어헬스케어학과) , 이송희 (을지대학교대학원 임상병리학과) , 이소립 (을지대학교대학원 BK21 프로그램 시니어헬스케어학과) , 이한 (을지대학교 임상병리학과) , 성호중 (을지대학교대학원 임상병리학과) , 강희규 (을지대학교대학원 임상병리학과) , 현성희 (을지대학교대학원 임상병리학과)

초록
AI-Helper

면봉을 사용한 샘플링 방법은 의학, 생태학, 생명공학, 법의학 및 오염 정도 모니터링 시스템과 같은 다양한 연구 분야에서 유용하다. 샘플링에서 채취 시약은 중요한 요소 중 하나이다. 시료를 장기간 보관해야 하는 경우에는 시료 채취 키트와 시료 보관 온도가 매우 중요하다. 이 연구의 목적은 별도의 배지 없이, 세 가지 채취 시약과 세 가지 보관 온도가 살아있는 세균의 생균 수에 미치는 영향을 확인하는 것이다. 대표적인 환경 세균으로 E. coli와 S. aureus를 선정하였다. 증류수(DW), 멸균된 인산염 완충액(PBS), Tris-EDTA (TE) 버퍼를 채취 시약으로 사용하여 샘플링한 후, 22, 4, -70℃에서 보관을 진행했다. 각 채취시약 및 보관 온도가 시료에 미치는 영향은 RLU와 CFU로 결과로 비교하였다. -70℃ 보관 온도와 TE 버퍼를 사용할 때 CFU와 RLU 값은 다른 조건에서보다 일정하게 유지되는 경향이 보였다. 따라서 이 연구는 시료가 채취 직후 -70℃에서 보관되어야 하며, 채취 시약으로 TE 버퍼와 함께 사용하는 것이 좋다는 것을 시사한다.

Abstract ▼ AI-Helper

Swabs are useful and common sampling tools in various research fields, such as medicine, ecology, biotechnology, forensic medicine, and pollutant monitoring systems. Collection reagents are one of the essential components in sampling. It is important to develop a sample collection kit and designate an appropriate storage temperature because samples need to be stored for a long time. The purpose of this study was to identify the effects of three collection reagents and three storage temperatures on the recovery of living bacteria without media. We selected Escherichia coli and Staphylococcus aureus as representative environmental bacteria. Distilled water (DW), phosphate buffered saline (PBS), and Tris-EDTA (TE) buffer were used as collection reagents and stored at 22℃, 4℃, and -70℃ after sampling. The results of using each collection reagent and storage temperature on the bacteria were compared using relative light units (RLU) and the number of colony forming units (CFU). When using -70℃ storage temperature and the TE buffer, the number of living bacteria and the RLU values remained constant. It is therefore recommended that the sample be stored at -70℃ immediately after collection and a TE buffer solution be used as the collection reagent.

주제어

표/그림 (3)

그림 Figure 1. Each bacteria were cultured on BHIA and incubated at 37°C for 24 h. After that, two colonies of E. coli and one colony of S. aureus from BHIA were incubated in LB broth for 24 h at 37°C. 5 mL of E. coli in LB broth and 5 mL of S. aureus in LB broth were mixed. With sterilized cotton, we smeared the bacterial solution on a desk. The samples were swabbed from the 5x5 cm area of the substrates for each condition.
그림 Figure 2. Bar graphs of RLU and CFU values of samples devided by storage temperatures. All resulting values were converted to logarithms. Figure A, C, E are signals detections of E. coli and S. aureus (Log RLU/mL±SD), Figure B, D, F are the number of colonies of E. coli and S. aureus (Log CFU/mL±SD). Each pattern represents the reagents.
그림 Figure 3. Scatterplot of ATP assay values (Log RLU/mL) and colony count (Log CFU/mL) of samples. Graph about correlation between RLU values and CFU values. All resulting values were converted to logarithms.

참고문헌 (29)

Jansson L, Akel Y, Eriksson R, Lavander M, Hedman J. Impact of swab material on microbial surface sampling. J Microbiol Methods. 2020;176:106006. https://doi.org/10.1016/j.mimet.2020.106006

상세보기
Panisello Yague D, Mihaljevic J, Mbegbu M, Wood CV, Hepp C, Kyman S, et al. Survival of Staphylococcus aureus on sampling swabs stored at different temperatures. J Appl Microbiol. 2021;131:1030-1038. https://doi.org/10.1111/jam.15023

상세보기
Zasada AA, Zacharczuk K, Wonznica K, Glowka M, Ziolkowski R, Malinowska E. The influence of a swab type on the results of point-of-care tests. AMB Express. 2020;10:46. https://doi.org/10.1186/s13568-020-00978-9

상세보기
You HS, Lee SH, Ok YJ, Kang HG, Sung HJ, Lee JY, et al. Influence of swabbing solution and swab type on DNA recovery from rigid environmental surfaces. J Microbiol Methods. 2019;161:12-17. https://doi.org/10.1016/j.mimet.2019.04.011

상세보기
Song YB, Ko SR, Kim CS, Kwak YS. Rapid microbiological assessment method by using ATP-bioluminescence in Ginseng powder. J Ginseng Res. 2001;25:127-129.
Turner DE, Daugherity EK, Altirer C, Maurer KJ. Efficacy and limitations of an ATP-based monitoring system. J AM Assoc Lab Anim Sci. 2010;49:190-195.
Kanoatov M, Krylov SN. Analysis of DNA in phosphate buffered saline using kinetic capillary electrophoresis. Anal Chem. 2016; 88:7421-7428. https://doi.org/10.1021/acs.analchem.6b02117

상세보기
Ross KS, Haites NE, Kelly KF. Repeated freezing and thawing of peripheral blood and DNA in suspension: effects on DNA yield and integrity. J Med Genet. 1990;27:569-570. https://doi.org/10.1136/jmg.27.9.569

상세보기
Lee HM, Yang JH, Gwon SY, Kang HG, Hyun SH, Lee JY, et al. Development of novel extraction reagents for analyzing dried blood spots from crime scenes. Forensic Sci Int. 2020;317:110531. https://doi.org/10.1016/j.forsciint.2020.110531

상세보기
Gosselt HR, Griffioen PH, van Zelst BD, Oosterom N, de Jonge R, Heil SG. Global DNA (hydroxy)methylation is stable over time under several storage conditions and temperatures. Epigenetics. 2021;16:45-53. https://doi.org/10.1080/15592294.2020.1786318

상세보기
Howlett SE, Castillo HS, Gioeni LJ, Robertson JM, Donfack J. Evaluation of DNA stable for DNA storage at ambient temperature. Forensic Sci Int Genet. 2014;8:170-178. https://doi.org/10.1016/j.fsigen.2013.09.003

상세보기
Saliba R, Zahar JR, El Allaoui F, Carbonnelle E, Lescat M. Impact of freeze/thaw cycles and single freezing at -80 degrees on the viability of aerobic bacteria from rectal swabs performed with the ESwab(TM) system. Diagn Microbiol Infect Dis. 2020;96:114895. https://doi.org/10.1016/j.diagmicrobio.2019.114895

상세보기
Delacour H, Van Cuyck H, Dubrous P, Soullie B, Leroy P, Koeck JL. Efficacy of a swab transport system in maintaining long-term viability of Staphylococcus aureus. Diagn Microbiol Infect Dis. 2009;65:345-346. https://doi.org/10.1016/j.diagmicrobio.2009.07.011

상세보기
Smith PW, Sayles H, Hewlett A, Cavalieri RJ, Gibbs SG, Rupp ME. A study of three methods for assessment of hospital environmental cleaning. Healthcare Infection. 2013;18:80-85. https://doi.org/10.1071/HI13001

상세보기
Kang HM, Eom YS, An HS, Kim CJ, Choi KH, Chung CI. Application of ATP Bioluminescence method for measurement of microbial contamination in raw meat, meat and dairy processing line. J Food Hyg Saf. 2000;15:252-255.
Romeo L, Lanza Cariccio V, lori R, Rollin P, Bramanti P, Mazzon E. The α-cyclodextrin/moringin complex: a new promising antimicrobial agent against Staphylococcus aureus. Molecules. 2018;23:2097. https://doi.org/10.3390/molecules23092097

상세보기
Al-Qadiri HM, Al-Alami NI, Al-Holy MA, Rasco BA, Using fourier transform infrared (FT-IR) absorbance spectroscopy and multivariate analysis to study the effect of chlorine-induced bacterial injury in water. J Agric Food Chem. 2008;56:8992-8997. https://doi.org/10.1021/jf801604p

상세보기
Aiken ZA, Wilson M, Pratten J, Epidemiology H. Evaluation of ATP bioluminescence assays for potential use in a hospital setting. Infect. Control Hosp Epidemiol. 2011;32:507-509. https://doi.org/10.1086/659761

상세보기
Sule P, Wadhawan T, Wolfe A, Pruss BJPn. Use of the BacTiter-Glo TM microbial cell viability assay to study bacterial attachment in biofilm formation. Promega Notes. 2008;99:19-21.
Hedman J, Akel Y, Jansson L, Hedell R, Wallmark N, Forsberg C, et al. Enhanced forensic DNA recovery with appropriate swabs and optimized swabbing techniqe. Forensic Sci Int Genet. 2021;53:102491. https://doi.org/10.1016/j.fsigen.2021.102491

상세보기
Verdon TJ, Mitchell RJ, van Oorschot RAH. Swabs as DNA collection devices for sampling different biological materials from different substrates. J Forensic Sci. 2014;59:1080-1089. https://doi.org/10.1111/1556-4029.12427

상세보기
Low SXZ, Aw ZQ, Loo BZL, Lee KC, Oon JSH, Lee CH, et al. Viability of Escherichia coli ATCC 8739 in nutrient broth, luriabertani broth and brain heart infusion over 11 weeks. Electronic physician. 2013;5:576-581. https://doi.org/10.14661/2013.576-581

상세보기
Calvert R, Hopkins H, Reilly M, Forsythe S. Caged ATP-an internal calibration method for ATP bioluminescence assays. Lett Appl Microbiol. 2000;30:223-227. https://doi.org/10.1046/j.1472-765x.2000.00703.x

상세보기
Song-im N. Benson S. Lennard C. Evaluation of different sampling media for their potential use as a combined swab for the collection of both organic and inorganic explosive residues. Forensic Sci Int. 2012;222:102-110. https://doi.org/10.1016/j.forsciint.2012.05.006

상세보기
Gibbs SG, Sayles H, Colbert EM, Hewlett A, Chaika O, Smith PW. Evaluation of the relationship between the adenosine triphosphate (ATP) bioluminescence assay and the presence of bacillus anthracis spores and vegetative cells. Int J Environ Res Public Health. 2014;11:5708-5719. https://doi.org/10.3390/ijerph110605708

상세보기
Salsgiver E, Bernstein D, Simon MS, Greendyke W, Jia H, Robertson A, et al. Comparing the bioburden measured by adenosine triphosphate (ATP) luminescence technology to contact plate-based microbiologic sampling to assess the cleanliness of the patient care environment. Infect Control Hosp Epidemiol. 2018;39:622-624. https://doi.org/10.1017/ice.2018.39

상세보기
La Duc MT, Dekas A, Osman S, Moissl C, Newcombe D, Venkateswaran K. Isolation and characterization of bacteria capable of tolerating the extreme conditions of clean room environments. Appl Environ Microbiol. 2007;73:2600-2611. https://doi.org/10.1128/AEM.03007-06

상세보기
Oz P, Arun OOJF, Health. Evaluating the performance of ATP bioluminescence method by comparison with classical cultural method. Food and Health. 2019;5:77-82. https://doi.org/10.3153/FH19008
Smith S, Morin PA. Optimal storage conditions for highly dilute DNA samples: a role for trehalose as a preserving agent. J Forensic Sci. 2005;50:1101-1108. https://doi.org/10.1520/JFS2004411

상세보기

저자의 다른 논문 :

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증