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Colonization of Pathogens in Earphones and Observation of Effective Sterilization Methods and Cycles
Biomed Sci Letters 2022;28:186-191
Published online September 30, 2022;  https://doi.org/10.15616/BSL.2022.28.3.186
© 2022 The Korean Society For Biomedical Laboratory Sciences.

Hyeokjin Kwon*, Myeongguk Jeong*, Shinjee Go**, Yeojin Kim**, Yein Kim**, Yeeun Kim**, Seungjun Roh**, Seonggwang Lee** and Go-Eun Choi†,***

Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Korea
Correspondence to: Go-Eun Choi. Department of Clinical Laboratory Science, College of Health Sciences, Catholic University of Pusan, Busan 46252, Korea.
Tel: +82-51-510-0563, Fax: +82-51-510-0568, e-mail: gechoi@cup.ac.kr
*Graduate student, **Ungraduate student, ***Professor.
Received September 19, 2022; Revised September 23, 2022; Accepted September 23, 2022.
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
 Abstract
The use of earphones has recently been widely used around the world. In currently, students wear earphones a lot in a daily life. The types of earphones are open-earphones, Canalphones, and headphones. Many students don't periodically to sterilization their earphones. Therefore, it can be an incubator that can induced ear infections. The objective of this study was to detect the pathogenic bacteria from the earphones used by the students. A total of 3 type earphones swabs were collected by sterile cotton swabs. The swabs were inoculated onto BHI agar and incubated aerobically 48 hour at 37꼦. 16s rRNA PCR, electrophoresis and sequencing were performed to confirm the identification of all the bacterial isolates. As a result, 24 pathogens were identified in sequencing. Three types of earphones were sterilized in three ways: ultraviolet (UV), 70% ethyl alcohol, and antibacterial wet tissue. If you use earphones for a long time without disinfecting them for a long time, it causes various diseases such as external ear infections. The findings of this study the users periodically to sterilization their respective earphones.
Keywords : Externa, Earphone, PCR, Electrophoresis, Sterilization
꽌 濡

理쒓렐 뱾뼱 샎옄 떆媛꾩쓣 蹂대궡뒗 臾명솕媛 以묓솕릺硫댁꽌 二쇰쓽 냼쓬쓣 쉶뵾븯湲 쐞븳 諛⑹븞쑝濡 씠뼱룿 궗슜씠 뒛뼱굹怨 엳떎. 듅엳, 泥냼뀈뱾쓽 씠뼱룿 궗슜 鍮덈룄媛 꽦씤쓽 궗슜 鍮덈룄蹂대떎 뜑 넂寃 굹궗쑝硫, 二쇰줈 뒪留덊듃룿씠굹 깭釉붾┸ PC瑜 씠슜븯뿬 씤꽣꽬 媛뺤쓽굹 쁺긽 벑쓣 留롮씠 떆泥븯뒗 寃껋쑝濡 議곗궗맂 諛 엳떎(Lee, 2018; Koo and Kim, 2020). 씠뼱룿쓣 궗슜븯뒗 궗엺뱾씠 젏李 利앷븯硫댁꽌 愿젴 吏덈퀝 삉븳 옄二 諛쒖깮븯뒗뜲 洹 以 몴쟻씤 吏덈퀝 以 븯굹媛 쇅씠룄뿼씠떎(Abbinay and Bharathi, 2012). 쇅씠룄뒗(Externa) 洹瑜 援ъ꽦븯뒗 遺遺 以묒뿉꽌 洹볥컮댁뿉꽌 怨좊쭑源뚯쓽 湲몄쓣 留먰븯硫, 쇅씠룄뿼 뿬윭 媛吏 썝씤쑝濡 씤빐 諛쒖깮릺뒗뜲 몴쟻쑝濡 쇅씠룄媛 꽭洹좎씠굹 怨고뙜씠 벑뿉 媛먯뿼릺뼱 뿼利앹씠 깮湲대떎. 듅엳 쇅씠룄뿼 뒿븳 뿬由꾩뿉 옒 깮湲곕뒗 吏덊솚씠硫, 닔쁺옣씠굹 紐⑹슃쓣 븳 썑 洹瑜 썑鍮꾨뒗 뻾쐞瑜 넻빐 뵾遺媛 빟빐吏硫댁꽌 꽭洹좎씠굹 怨고뙜씠뿉 쓽빐 媛먯뿼씠 돺寃 諛쒖깮븷 닔 엳떎(Nichols, 1999; Wiegand et al., 2019). 쇅씠룄뿼쓽 빟 90% 씠긽 꽭洹좎뿉 쓽빐 쑀諛쒕릺硫(Roland and Stroman, 2002), 끃냽洹(Pseudomonas aeruginosa) 22~62%, 솴깋룷룄긽援ш퇏(Staphylococcus aureus) 11~34% 몢 媛쒖쓽 洹 醫낆씠 媛옣 쓷븯寃 쇅씠룄뿼쓣 쑀諛쒗븳떎(Ijaz et al., 2014; Rosenfeld et al., 2014; Musso and Crews, 2016). 洹 쇅쓽 빟 10%쓽 洹좎쑝濡쒕뒗 늻猷⑷낡뙜씠냽(Aspergillus) 80~90%媛 엳떎(Boustred, 1999; Sander, 2001). 쇅씠룄뿼쓽 珥덇린 利앹긽 媛젮利앷낵 븿猿 誘몃명븳 넻利앹쓣 룞諛섑븯硫, 씠썑뿉뒗 洹뱀떖븳 넻利앹씠 룞諛섎맂떎(Wiegand et al., 2019). 씠濡 씤빐 떇궗뿉 諛⑺빐瑜 二쇨퀬 닔硫댁씠굹 蹂댄뻾뿉 뼱젮쓣 룞諛섑븳떎. 쇅씠룄뿼쓽 移섎즺 諛⑸쾿 洹瑜 源⑤걮븯寃 냼룆븯怨, 빆깮젣 吏꾪넻젣 벑쓽 빟臾쇰줈 移섎즺븯뿬빞 븳떎. 쇅씠룄뿉 怨좊쫫 二쇰㉧땲媛 삎꽦맂 寃쎌슦뿉뒗 씠瑜 젅媛쒗빐 怨좊쫫쓣 젣嫄고빐빞 븯硫 쇅씠룄뿼 泥냼뒗 諛섎뱶떆 씠鍮꾩씤썑怨 쓽궗뿉 쓽빐 議곗떖뒪읇寃 뻾빐졇빞 븳떎. 쇅씠룄뿼 벑 洹 吏덊솚쓣 삁諛⑺븯젮硫 넀怨 洹, 씠뼱룿 벑쓣 二쇨린쟻쑝濡 냼룆븯뿬빞 븳떎(Sander, 2001). 씠뼱룿쓣 궗슜븯뒗 궗엺 젏李 利앷븯뒗 諛섎㈃, 쇅씠룄뿼 벑 愿젴 吏덈퀝뿉 븳 쐞뿕꽦怨 냼룆 諛⑸쾿뿉 愿빐 븣젮吏 諛붽 嫄곗쓽 뾾뼱 냼룆쓽 븘슂꽦쓣 씤떇떆궎湲 쐞빐 蹂 뿰援щ 吏꾪뻾븯떎. 蹂 뿰援ъ뿉꽌뒗 씠뼱룿 3媛吏 醫낅쪟瑜 궗슜븯뿬 뼱뼚븳 蹂묒썝洹좎씠 엳뒗吏 솗씤븯怨 옄쇅꽑(UV), 70% Ethyl alcohol, 빆洹 臾쇳떚뒋瑜 씠슜븯뿬 냼룆 쟾 썑瑜 鍮꾧탳 遺꾩꽍빐 냼룆쓽 븘슂꽦쓣 솗씤븯怨좎옄 븳떎(Madigan et al., 2006; Oughton et al., 2009; Buonanno et al., 2017).

옱猷 諛 諛⑸쾿

떆猷 닔吏 諛 DNA 異붿텧

蹂 뿰援ъ뿉꽌 궗슜맂 씠뼱룿 쇅쓽쓽 젒珥됰㈃뿉 뵲씪 醫낅쪟瑜 굹늻뼱 떎뿕븯떎. 꽑젙맂 3媛吏 醫낅쪟쓽 씠뼱룿 삤뵂 씠뼱룿(Open-earphones), 而ㅻ꼸 씠뼱룿(Canalphones), 뿤뱶룿(Headphones)쑝濡 궗슜븯쑝硫, 븯猷 룊洹 3떆媛 씠긽 궗슜븯뒗 븰깮 긽쓽 씠뼱룿쓣 닔吏묓븯떎. 떆猷 닔吏묒 硫멸퇏 硫대큺쑝濡 씠뼱룿 몴硫댁쓣 湲곸 썑 BHI agar뿉 젒醫낇븯떎. 諛곗뼇 議곌굔 37℃, 48떆媛 諛곗뼇븯쑝硫, 諛곗뼇 썑 珥 24媛쒖쓽 떒씪 吏묐씫쓣 닔吏묓븯떎. 닔吏묓븳 떒씪 吏묐씫 DNA 異붿텧뿉 궗슜븯떎. DNA 異붿텧 Kit (AccuPrep® Genomic DNA Extraction Kit - 3032) 젣議곗궗쓽 吏떆뿉 뵲씪 궗슜븯떎.

PCR, 쟾湲곗쁺룞

16S rRNA 쑀쟾옄 꽌뿴쓽 PCR 利앺룺 Bacteria primer (Table 1)瑜 궗슜븯떎. PCR 怨쇱젙(Marchesi et al., 1998)쓣 李멸퀬 諛 蹂삎븯뿬 諛⑸쾿쓣 궗쁺븯떎. Initial denaturation 95℃뿉꽌 10遺 denaturation 95℃ 30珥, annealing 54℃ 30珥, extension 72℃ 30珥, 32 Cycle 利앺룺 썑 Final extension 72℃ 5遺 룞븞 떎떆븯떎. 1% Agarose gel (Lonza, Basel, Swiss)뿉 NEOgreen (CELLGENTEK, Deajeon, Korea) 떆빟쓣 샎빀 썑 Mupid-2plus (Takara, Kusatsu, Japan)瑜 궗슜븯뿬 떎떆븯怨, ChemiDoc XRS+ (Bio-Rad, CA, USA)瑜 씠슜븯뿬 DNA Band瑜 솗씤븯떎. 쟾湲곗쁺룞 怨쇱젙(Southern, 1975)쓣 李멸퀬 諛 蹂삎븯뿬 떎떆븯떎. 뿼湲곗꽌뿴 遺꾩꽍 Macrogen (Seoul, Korea)뿉 쓽猶고븯뿬 룞젙븯떎.

Primer sequences to amplify 16s rRNA gene

Primer name Sequence Amplicon (bp) References
Forward 27F 5'-AGAGTTTGATCCTGGCTCAG-3' 1,504 (Lane et al., 1985)
Reverse 1492r 5'-GGTTACCTTGTTACGACTT-3'


씠뼱룿 냼룆

씠뼱룿 냼룆 諛⑸쾿 씤泥댁뿉 쑀빐븯吏 븡怨 쓷엳 궗슜릺硫, 湲곌린뿉 넀긽쓣 媛븯吏 븡뒗 諛⑸쾿쓣 궗슜븯떎. 媛 떎뿕援 蹂꾨줈 옄쇅꽑(UV) 泥섎━, 70% Ethyl alcohol 泥섎━, 빆洹 臾쇳떚뒋 泥섎━濡 굹늻뼱 냼룆븯쑝硫 옄쇅꽑(UV) 냼룆 254 nm 뙆옣쑝濡 10遺, 20遺, 30遺꾩쑝濡 吏꾪뻾븯쑝硫, 빆洹 臾쇳떚뒋(BIOMAX, Seoul), 70% Ethyl alcohol쓣 씠슜븯뿬 씠뼱룿 몴硫댁쓣 떐븯떎. 씠 썑 궡洹 슚怨쇰 솗씤븯湲 쐞빐 냼룆 썑 3떆媛 룞븞 李⑹슜븯怨, 떆猷 梨꾩랬뒗 硫멸퇏 硫대큺쓣 씠슜븯뿬 씠뼱룿 몴硫댁뿉꽌 梨꾩랬븯뿬 BHI agar뿉 젒醫낇뻽쑝硫, 洹 썑 CFU 닔瑜 痢≪젙븯떎.

寃 怨

떆猷 닔吏 諛 PCR

3媛吏 醫낅쪟쓽 씠뼱룿(Open-earphones, Canalphones, Headphones) 몴硫댁쓣 硫멸퇏 硫대큺쑝濡 湲곸뼱 BHI agar뿉 젒醫낇븯뿬 37℃, 48떆媛 諛곗뼇 썑 24媛쒖쓽 떒씪 吏묐씫쓣 닔吏묓븯쑝硫, 닔吏묓븳 떆猷뚮뒗 16s rRNA PCR뿉 닔뻾븯떎.

쟾湲곗쁺룞 諛 뿼湲곗꽌뿴 遺꾩꽍

利앺룺맂 PCR 궛臾쇱쓣 쟾湲곗쁺룞 븳 썑(Fig. 1), Macrogen뿉 뿼湲곗꽌뿴 遺꾩꽍쓣 쓽猶고븯떎. 뿼湲곗꽌뿴 遺꾩꽍 寃곌낵뒗 NCBI BLAST (National Center for Biotechnology information Basic Local Alignment Search Tool) 봽濡쒓렇옩쓣 넻빐 뿼湲곗꽌뿴쓽 쑀궗룄瑜 遺꾩꽍븯뿬 24媛쒖쓽 洹 룞젙쓣 솗씤븯떎. Staphylococcus epidermidis 洹좎쥌씠 12嫄 Staphylococcus capitis 洹좎쥌 5嫄, Staphylococcus caprae 洹좎쥌 2嫄, Staphylococcus hominis subsp. Novobiosepticus 洹좎쥌 2嫄 洹몃━怨 Kocuria marina, Micrococcous yunnanensis, Bacillus velezensis 洹좎쥌씠 媛 1嫄댁뵫 遺꾨━릺뿀떎(Table 2).

Sequencing results of identified pathogens in 3 types of earphones

Type No. Binominal nomenclature 16s rRNA Per. Ident
Open earphones 1 Staphylococcus capitis 100%
2 Staphylococcus hominis subsp. Novobiosepticus 99.8%
3 Staphylococcus hominis subsp. Novobiosepticus 99.6%
4 Staphylococcus epidermidis 100%
5 Staphylococcus epidermidis 99.9%
Canalphones 1 Staphylococcus epidermidis 99.7%
2 Staphylococcus epidermidis 99.9%
3 Staphylococcus epidermidis 99.8%
4 staphylococcus caprae 98%
5 Staphylococcus epidermidis 99.82%
6 Staphylococcus epidermidis 99.9%
7 Staphylococcus epidermidis 100%
8 Staphylococcus epidermidis 99.56%
9 Staphylococcus epidermidis 100%
10 Micrococcus yunnanensis 100%
11 Staphylococcus capitis 100%
12 Staphylococcus capitis 100%
13 Staphylococcus epidermidis 100%
14 Bacillus velezensis 100%
15 Staphylococcus capitis 100%
16 Staphylococcus caprae 100%
Headphones 1 Staphylococcus epidermidis 100%
2 Staphylococcus capitis subsp. urealyticus 100%
3 Kocuria marina 99.8%


Fig. 1. PCR gel electrophoresis showing 16S rRNA gene bands (at 1,500 bp) of the isolated bacterial groups. Samples were collected bacteria from earphones and then 1% agarose gel electrophoresis analysis of 16s rRNA gene amp lified. Lane M, 1 kb DNA ladder; lanes 1 to 5 are Open earphones DNA bands, and lanes 6 to 21 are Canalphones DNA bands. The last 22nd to 24th lanes are Headphones DNA bands.

씠뼱룿 냼룆

옄쇅꽑(UV), 70% Ethyl alcohol, 빆洹 臾쇳떚뒋 3媛吏 諛⑸쾿쑝濡 냼룆븳 寃곌낵 옄쇅꽑(UV) 10遺꾧낵 20遺꾩뿉꽌 냼룆 슚怨쇰 蹂 닔 뾾뿀쑝硫, 30遺꾨꽣 냼룆쓽 슚怨쇰 蹂 닔 엳뿀떎(Table 3). 빆洹 臾쇳떚뒋, 70% Ethyl alcohol 냼룆 諛⑸쾿 옄쇅꽑 10遺, 20遺 蹂대떎 슚怨쇰 蹂 닔 엳뿀떎(Table 3). 냼룆쓽 슚怨쇰뒗 옄쇅꽑(UV) 냼룆 30遺꾩씠 媛옣 醫뗭븯쑝硫 떎쓬 70% Ethyl alcohol, 빆洹 臾쇳떚뒋 닚쑝濡 냼룆뿉 슚怨쇨 엳뿀떎. 옄쇅꽑쓣 10遺꾧컙 끂異쒖떆耳곗쓣 떆 삤뵂삎 씠뼱룿 172 CFU/mL뿉꽌 45 CFU/mL濡 26.1% 궡洹좊릺뿀怨, 而ㅻ꼸삎 씠뼱룿 TNTC (Too Numerous To Count)뿉꽌 TNTC濡 0%, 뿤뱶룿 51 CFU/mL뿉꽌 4 CFU/mL濡 7.84% 궡洹좊릺뿀떎. 20遺 끂異쒖떆耳곗쓣 븣뿉뒗 삤뵂삎 씠뼱룿 1,208 CFU/mL뿉꽌 9 CFU/mL濡 99.25%, 而ㅻ꼸삎 씠뼱룿 TNTC뿉꽌 25 CFU/mL濡 99%, 뿤뱶룿 542 CFU/ml뿉꽌 8 CFU/mL濡 98.5% 궡洹좊릺뿀떎. 30遺 끂異쒖떆耳곗쓣 븣뿉뒗 3醫낅쪟 씠뼱룿 紐⑤몢 100% 궡洹 슚怨쇰 蹂 닔 엳뿀떎. 70% Ethyl alcohol쑝濡 씠뼱룿 몴硫댁쓣 떐븯쓣 븣뿉 삤뵂삎 씠뼱룿 398 CFU /mL뿉꽌 0 CFU/mL濡 100%, 而ㅻ꼸삎 씠뼱룿 TNTC뿉꽌 3 CFU/mL濡 99%, 뿤뱶룿 164 CFU/mL뿉꽌 3 CFU/mL濡 98.17% 궡洹좎씠 릺뿀쑝硫, 빆洹좊Ъ 떚뒋瑜 씠슜빐꽌 씠뼱룿 몴硫댁쓣 떐븯쓣 븣뿉뒗 삤뵂삎 씠뼱룿 326 CFU/mL뿉꽌 3 CFU/mL濡 99.08%, 而ㅻ꼸삎 씠뼱룿 789 CFU/mL뿉꽌 17 CFU/mL濡 97.85%, 뿤뱶룿 141 CFU/mL뿉꽌 2 CFU/mL濡 98.58% 궡洹좊릺뿀떎.

The results of the CFU counts after the sterilization

Type UV
10 min 20 min 30 min
Before (CFU/mL) After (CFU/mL) Sterilization rates (%) Before (CFU/mL) After (CFU/mL) Sterilization rates (%) Before (CFU/mL) After (CFU/mL) Sterilization rates (%)
Open earphones 172 45 26.1% 1,208 9 99.25% 544 0 100%
Canalphones TNTC TNTC 0% TNTC 25 99% 1,592 0 100%
Headphones 51 4 7.84% 542 8 98.5% 327 0 100%
Type 70% Ethyl alcohol Antibacterial wet tissue
Before (CFU/mL) After (CFU/mL) Sterilization rates (%) Before (CFU/mL) After (CFU/mL) Sterilization rates (%)
Open earphones 398 0 100% 326 3 99.08%
Canalphones TNTC 3 99% 789 17 97.85%
Headphones 164 3 98.17% 141 2 98.58%

TNTC: Too Numerous To Count


寃곕줎 諛 怨좎같

씠뼱룿 냼룆 쇅씠룄뿼씠굹 뿼利앹쓣 삁諛⑺븯怨 蹂묒썝媛먯뿼쓣 愿由ы븯뒗뜲 엳뼱 媛꾨떒븯硫댁꽌룄 媛옣 以묒슂븳 諛⑸쾿씠떎. 슚怨쇱쟻씤 냼룆쓣 븯湲 쐞빐꽌뒗 냼룆 諛⑸쾿, 떆媛꾩씠 以묒슂븯硫, 씠뼱룿 湲곌린 넀긽씠 뾾뼱빞 븳떎. 쇅씠룄뿼 뿬윭 媛吏 썝씤쑝濡 쇅씠룄뿉 꽭洹좎씠굹 怨고뙜씠 벑뿉 媛먯뿼릺뼱 뿼利앹씠 깮湲곕뒗 寃껋쓣 留먰븳떎. 쇅씠룄쓽 빟 90% 씠긽 꽭洹좎뿉 쓽빐 깮湲곕뒗 寃껋씠硫 떖븷 寃쎌슦 븙꽦 쇅씠룄뿼씠 諛쒖깮븷 닔 엳떎. 蹂 뿰援щ뒗 씠뼱룿쓣 넻븳 쇅씠룄뿼 諛 뿼利앹뿉 떖媛곸꽦怨 븞쟾꽦 솗蹂대 쐞빐 씠뼱룿뿉 엳뒗 蹂묒썝洹좎쓣 李얘퀬 삁諛⑷낵 씠뼱룿 냼룆쓽 以묒슂꽦쓣 븣由ш퀬옄 븯떎.

쓷엳 궗슜븯뒗 씠뼱룿 3醫낅쪟(Open earphones, Canalphones, Headphones)瑜 긽쑝濡 뿼湲곗꽌뿴 遺꾩꽍 寃곌낵 Staphylococcus epidermidis 洹좎쥌씠 12嫄 Staphylococcus capitis 洹좎쥌 5嫄, Staphylococcus caprae 洹좎쥌 2嫄, Staphylococcus hominis subsp. Novobiosepticus 洹좎쥌 2嫄 洹몃━怨 Kocuria marina, Micrococcous yunnanensis, Bacillus velezensis 洹좎쥌씠 媛 1嫄댁뵫 遺꾨━릺뿀떎. S. epidermidis, S. capitis, S. caprae, S. hominis 洹좎쥌 Coagulase Negative Species (CoNS)씠硫, 뵾遺 諛 젏留됱뿉 꽌떇븯뒗 긽옱洹좎씠떎. 嫄닿컯븳 궗엺뿉寃뚮뒗 吏덈퀝쓣 빞湲고븯吏 븡吏留, 硫댁뿭젰씠 븯맂 궗엺뿉寃 蹂듬쭑뿼, 씤怨 뙋留 떖궡留됱뿼, 닔留됱뿼, 怨⑥닔뿼 벑 뿬윭 吏덈퀝쓣 빞湲고븷 닔 엳떎. K. marina 뵾遺 援ш컯뿉 꽌떇븯뒗 洹좎쥌씠硫, 鍮꾨퀝썝꽦쑝濡 媛꾩<릺吏留 슂濡쒓컧뿼, 떞궘뿼, 媛곷쭑뿼 벑 씪遺 吏덈퀝쓣 쑀諛쒗븷 닔 엳떎.

닔吏묐맂 誘몄깮臾쇱쓣 긽쑝濡 궡洹 슚怨쇰 솗씤븯湲 쐞빐 옄쇅꽑(UV), 70% Ethyl alcohol, 빆洹 臾쇳떚뒋 3媛吏 諛⑸쾿쑝濡 냼룆븯떎. 떎뿕 쟾怨 썑쓽 誘몄깮臾 遺꾪룷룄瑜 鍮꾧탳 遺꾩꽍븯湲 쐞빐, 3醫낅쪟 씠뼱룿쓣 냼룆 쟾怨 냼룆 썑 3떆媛 룞븞 李⑹슜븳 씠뼱룿쑝濡 洹몃9솕 븯뿬 鍮꾧탳 遺꾩꽍븯떎. 떆猷 梨꾩랬뒗 硫멸퇏 硫대큺쓣 씠슜븯뿬 씠뼱룿 몴硫댁뿉꽌 梨꾩랬븯뿬 BHI agar뿉 젒醫낇뻽쑝硫, 洹 썑 CFU 닔瑜 痢≪젙븯떎. 洹 寃곌낵 옄쇅꽑쓣 10遺꾧컙 끂異쒖떆耳곗쓣 떆 삤뵂삎 씠뼱룿 26.1% 궡洹좊릺뿀怨, 而ㅻ꼸삎 씠뼱룿 0%, 뿤뱶룿 7.84% 궡洹좊릺뿀떎. 20遺 끂異쒖떆耳곗쓣 븣뿉뒗 삤뵂삎 씠뼱룿 99.25%, 而ㅻ꼸삎 씠뼱룿 99%, 뿤뱶룿 98.5% 궡洹좊릺뿀떎. 30遺 끂異쒖떆耳곗쓣 븣뿉뒗 3醫낅쪟 씠뼱룿 紐⑤몢 100% 궡洹 슚怨쇰 蹂 닔 엳뿀떎. 70% Ethyl alcohol쑝濡 씠뼱룿 몴硫댁쓣 떐븯쓣 븣뿉 삤뵂삎 씠뼱룿 100%, 而ㅻ꼸삎 씠뼱룿 99%, 뿤뱶룿 98.17% 궡洹좎씠 릺뿀쑝硫, 빆洹 臾쇳떚뒋瑜 씠슜빐꽌 씠뼱룿 몴硫댁쓣 떐븯쓣 븣뿉뒗 삤뵂삎 씠뼱룿 99.08%, 而ㅻ꼸삎 씠뼱룿 97.85%, 뿤뱶룿 98.58% 궡洹좊릺뿀떎.

寃곕줎쟻쑝濡 씠뼱룿 냼룆 쟾怨 썑 李⑥씠瑜 蹂댁뿬二쇰ʼn, 냼룆 쟾 씠뼱룿뿉꽌 뿬윭 誘몄깮臾쇱씠 梨꾩랬맂 寃껋쓣 솗씤븷 닔 엳뿀떎. 씠뼱룿쓣 옣떆媛 李⑹슜븷 寃쎌슦 洹볦냽 솚湲곕 뼱졄寃 븯뿬, 洹볦냽 뒿룄瑜 넂씠怨 誘몄깮臾 利앹떇쓽 썝씤씠 맂떎. 듅엳, 삤뵂삎 씠뼱룿怨 떖由 而ㅻ꼸삎 씠뼱룿쓽 寃쎌슦 李⑹슜떆 쇅씠룄 吏곸젒 젒珥됲븯湲 븣臾몄뿉, 二쇨린쟻쑝濡 냼룆븯湲곕 沅뚯옣븳떎.

떆猷 梨꾩랬 諛⑸쾿뿉꽌 뿰援ъ옄 븳 紐낆씠 룞씪븳 遺쐞뿉꽌 諛섎났븯뿬 寃泥대 梨꾩랬븯뜑씪룄 臾몄瑜대뒗 媛뺣룄굹 梨꾩쭛 遺쐞媛 씪移섑븯吏 븡쓣 媛뒫꽦씠 엳뿀떎. 븯吏留 蹂 뿰援 寃곌낵뿉꽌 궗슜븳 3媛吏 냼룆 諛⑸쾿 솚寃쎄由щ 泥댄븯湲 쐞븿씠 븘땲씪 씠뼱룿 궗슜옄뱾뿉寃 醫뗭 젣븞씠 맆 寃껋쑝濡 궗猷뚮맂떎.

ACKNOWLEDGEMENT

None.

CONFLICT OF INTEREST

No potential conflict of interest relevant to this article was reported.

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