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1 Ann Clin Microbiol Vol. 19, No. 1, March, pissn eissn Prevalence and Antimicrobial Susceptibility of Genital Mycoplasmataceae in Korean Women: Correlation between Phenotypic Test and Resistance Genes Jiyoung Chang 1, Jin Kyung Yu 1, Changeun Song 1, In Yang Park 2, Yeon-Joon Park 1 Departments of 1 Laboratory Medicine, 2 Obstetrics and Gynecology, College of Medicine, The Catholic University of Korea, Seoul, Korea Background: While 7.6% of cultured genital Mycoplasmataceae was identified as Ureaplasma urealyticum, most of them were Ureaplasma parvum (8.3%). This is the first study differentiating between these two species. We investigated the prevalence and antimicrobial resistance of genital Mycoplasmataceae in Korean women. Methods: A total of 15 specimens submitted to the laboratory for culture of M. hominis and Ureaplasma spp. were included. Detection and antimicrobial susceptibility tests were performed with the Mycoplasma IST2 kit (biomérieux, France). The identification of Ureaplasma spp. was performed by PCR, and mutations in drug resistance genes were investigated by PCR and sequencing. Results: In total, 66 specimens (44.%) were positive for genital Mycoplasmatacea: U. parvum, 53 (8.3%); U. urealyticum, 5 (7.6%); M. hominis, 2 (3.%); mixed infection, 6 (9.1%). Susceptibilities of Ureaplasma spp. to erythromycin, azithromycin, clarithromycin, and doxycycline were 86.%, 8.7%, 98.2%, and 94.7%, respectively. The susceptibility of Ureaplasma spp. to ofloxacin and ciprofloxacin was 47.4% and 17.5%, respectively. The S83L mutation was found in the ParC subunit of the ofloxacin-resistant (5/7, 71.4%) and the ciprofloxacin-resistant isolates (7/14, 5.%). One M. hominis isolate showed resistance to erythromycin, azithromycin, and clarithromycin but susceptibility to josamycin, pristinamycin, fluoroquinolones, and tetracyclines. Conclusion: The prevalence of genital Mycoplasmataceae in Korean women was 44.%; most of them were identified as U. parvum. As more than 1% of Ureaplasma spp. showed non-susceptibility to erythromycin and azithromycin (15.5%, 2.7%), a susceptibility test is needed prior to use of these antibiotics. Further study is needed about the clinical features of infections caused by U. urealyticum vs. U. parvum and their associated resistance mechanisms. (Ann Clin Microbiol 216;19:13-19) Key Words: Antimicrobial susceptibility, Mycoplasma IST2 kit, Ureaplasma parvum, Ureaplasma urealyticum INTRODUCTION Ureaplasma와 mycoplasma는세포벽이없는균속으로독립적인증식이가능한가장작은미생물체이다 [1]. 이들은 Mycoplasmataceae과에속하나, Ureaplasma 균속은 urea를가수분해하는능력이있어 Mycoplasma 균속과구별된다 [2]. Ureaplasma 와 mycoplasma는건강성인의비뇨생식기에서도분리되나, 요도염, 골반염, 불임, 자연유산, 조기분만, 조기진통, 조기양막파수, 저체중아, 조산아의호흡기감염, 패혈증등의원인으로여겨지고있다 [1-3]. Ureaplasma 에는두가지의뚜렷한변종이알 려져있는데, biovar 2 (Ureaplasma urealyticum) 은상기질환들을일으키나 biovar 1 (Ureplasma parvum) 의역할은뚜렷하지않다 [4,5]. 그러나 Martínez 등 [6] 의연구에서는두 biovars 모두양막강내침입하여임신기합병증과관련이있다고하였고 De Francesco 등 [7] 은생식기감염증상이있는여자환자의 44% 에서 U. parvum serovar 3/14가발견되었으며, Kong 등 [8] 은임신부질도말검체 87% 에서 U. parvum이동정되었고이는임신시합병증과관련이있었다고하였다. 한편 Xiao 등 [2] 의연구에따르면변종들의감별이임상적으로 ureaplasma 병원성을구별하는데도움을주지못하며감염된숙주의특이적면역 Received 1 November, 215, Revised 12 January, 216, Accepted 27 January, 216 Correspondence: Yeon-Joon Park, Department of Laboratory Medicine, College of Medicine, The Catholic University of Korea, Seoul St. Mary s Hospital, 222 Banpo-daero, Seocho-gu, Seoul 6591, Korea. (Tel) , (Fax) , ( ) yjpk@catholic.ac.kr c The Korean Society of Clinical Microbiology. This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. 13

2 14 Ann Clin Microbiol 216;19(1):13-19 반응에따라감염양상이다르게나타날것이라고하였다. Ureaplasma와 mycoplasma는세포벽이없어베타락탐 (βlactam) 제제에의해억제되지않고, 단백질합성억제에작용하는마크로라이드 (macrolides), 테트라사이클린 (tetracycline) 또는 DNA복제를억제하는퀴놀론 (quinolone) 등에감수성을보인다. 산모의경우생식기마이코플라즈마감염은조기분만위험이있으므로조기양막박리시주산기를연장하고그와관련된합병증과신생아감염을줄이기위해항균제투여가필요하다. 이때주로마크로라이드계열의약제가쓰이며, 마크로라이드에내성일경우독시싸이클린 (doxycycline) 이사용된다. 이러한마이코플라즈마의배양및감수성검사방법은매우까다로우므로대부분의임상검사실에서는상품화된 kit를사용한다. 본검사실에서는 Mycoplasma IST2 kit (biomérieux, Marcy l Etoile, France) 를사용하는데, 이검사법에서는 U. urealyticum과 U. parvum이모두자라므로이들의감별이필요하다. 그리고마이코플라즈마의 CLSI에서제안하는항생제감수성검사방법과 Mycoplasma IST2 kit (biomérieux) 에서사용되는항균제농도, 균액농도등의차이로실험방법이다르고설정된항생제별기준 (breakpoint) 또한다르다 [9]. Ureaplasma spp. 의항균제내성은마크로라이드의경우항균제작용부위의변이, 즉 23S rrna의 erythromycin 결합부위의유전자변이 (A266G) 가리보좀단백질 L4의변이 [A28C (T7P) or insertion] 또는 L22 부위의변이와동반되어일어나며 [1,11] 테트라싸이클린은내성유전자인 tet(m) 또는 tet(m) transposon의 marker인 int-tn에의한 ribosomal protection에의한다 [12]. 퀴놀론의경우 Type II topoisomerases인 DNA gyrase 와 topoisomerase IV를전사하는 gyra (A2399T, C31A), parc (G244A, C248T, G259A, T8A), pare (G1369A, C149T, C5T, A128T) 유전자의변이에의해초래되는아미노산의변이 gyra (N8I, Q14K), parc (D82N, S83L, E87K, V3E), pare (A457T, S47L, A2V, D427V) 가관여한다 [1,13]. 본연구에서는배양된 ureaplasma 중병원성이보고된 U. urealyticum의비율을알아보고, kit를이용한내성검사결과를내성유전자의변이와비교하고자하였다. MATERIALS AND METHODS 1. 대상 214년 1월부터 214년 11월까지서울성모병원진단검사의학과에 Ureaplasma/Mycoplasma hominis 배양검사가의뢰된 15개의검체들 ( 질도말검체 142, 자궁경부도말검체 5, 양수 3) 을대상으로하였고대상군의나이분포는중앙값 31세로최소 19세부터최대 49세사이였으며그중 118명이산모였다. 2. Mycoplasma IST kit 를이용한 Ureaplasma 배양동정및항생제감수성검사 Ureaplasma의배양동정과항생제감수성검사를위하여 Mycoplasma IST2 (biomérieux) 를사용하였다. 이를간단히설명하면다음과같다. 환자로부터수송배지에얻어진도말검체를 kit 내에있는시약인 R1배지에접종한후 3 ml를덜어 R2배지에넣고 vortex하여 lyophilized pellet이모두용해될때까지섞는다. 스트립의 22웰에혼합된용액을 55 μl씩분주한후파라핀오일을 2방울씩떨어뜨린다. 37 o C에서 24-48시간배양후각웰에있는용액의색변화로판독한다. 배지가적색이나오렌지색으로바뀌었을때생식기의마이코플라즈마가성장한것이며, 배지가노랑색일경우생식기의마이코플라즈마의성장이없는것을의미한다. 스트립은 M. hominis와 U. urealyticum의배양유무, 검체의균배양농도 ( 1 4 CFU), 항생제감수성 ; doxycycline (DOT), josamycin (JOS), ofloxacin (OFL), erythromycin (ERY), tetracycline (TET), ciprofloxacin (CIP), azithromycin (AZI), clarithromycin (CLA) 그리고 pristinamycin (PRI) 을판독할수있도록되어있고이는제조사의지침을따라하였다. 항생제별 breakpoint 는다음과같고이에따라항생제감수성을판단하였다. [ureaplasma (mg/l): erythromycin S 1, R 4; azithromycin S.12, R 4; clarithromycin S 1, R 4; josamycin S 2, R 8; ciprofloxacin S 1, R 2 ; ofloxacin S 1, R 4; tetracycline, doxycycline S 4, R 8; pristinamycin S 2 mycoplasma (mg/l): erythromycin S 1, R 4; azithromycin S.12, R 4; clarithromycin S 1, R 4; josamycin S 2, R 8; ciprofloxacin S 1, R 2; ofloxacin S 1, R 4; tetracycline, doxycycline S 4, R 8; pristinamycin S 2] 3. DNA 추출과 Ureaplasma 동정을위한 PCR 검사 7 o C에냉동시킨배양용액을해동한후전체 1 ml의용액을 12, revolutions per minute (RPM) 에서 5분간원심분리한후상청액을버리고침사를.1 M의 phosphate-buffered saline (PBS) 5 μl로씻어낸후 12, RPM에서 5분간원심분리하였다. DW 용액으로재부유시킨후 o C water bath에서 1 분간끓인다. 용액을식힌후 12, RPM에서 5분간원심분리후상청액을 PCR template로사용하였다. U. parvum, U. urealyticum 그리고 M. hominis 를구별하여동정하기위해 Stellrecht 등 [14] 이제안한방법을이용하였다. 4. 염기서열분석을통한항생제내성유전자변이검사마크로라이드계열항생제내성표현형의유전자변이를검사하기위하여마크로라이드비감수성균주에대하여 23S rrna domain II, domain V, 16S rrna L4 또는 L22 리보좀단백질유전자부위염기서열분석을시행하였고 [1,15] 이를마크

3 Jiyoung Chang, et al. : Prevalence and Antimicrobial Susceptibility of Genital Mycoplasmataceae 15 로라이드감수성 1개의균주와비교하여분석하였다. 테트라사이클린계열항생제비감수성균주에대하여 tet(m) 유전자와 tet(m) transposon의 marker인 int-tn 존재유무를 PCR로분석하였다 [16]. 퀴놀론계열항생제와관련된내성유전자 gyra,, parc, pare 변이를검사하기위하여퀴놀론내성균주의염기서열분석을시행하였다 [1]. 5. 통계수집된자료는 MedCalc, version (MedCalc Software, Mariakerke, Belgium) 을이용하여균농도별항생제감수성률비교를 Chi-square test로분석하였다. P값이.5 미만인경우통계학적으로의미가있는것으로해석하였다. RESULTS 1. Mycoplasma IST2 kit의배양결과와항생제감수성양상결과 Mycoplamsma IST2 kit (biomérieux) 를사용하여배양한결과균양성률을살펴보면전체생식기마이코플라즈마양성률은 44.% (66/15) 으로그중 Ureaplasma spp. 농도 1 4 이상은 17 검체 (11.3%), Ureaplasma spp. 농도 1 4 이하는 41검체 (27.3%), M. hominis는 1검체 (.7%), U. parvum과 M. hominis 의복합감염은 6검체 (4.%) 였고나머지 1검체 (.7%) 는판독이불가능하였다. PCR을이용하여균종동정한결과전체양성검체 66검체중 53검체에서 U. parvum (8.3%) 으로동정되었고 5검체에서 U. urealyticum (7.6%), 2검체에서 M. hominis (3.%), 6검체에서 U. parvum과 M. hominis 의복합감염 (9.1%) 으로동정되었다. Mycoplasma IST2 kit (biomérieux) 와 PCR과의일치율을살펴보면 Ureaplasma spp. 가 1 4 이상으로배양된 17검체중 16 검체가 PCR결과와일치하였고그중 14검체가 U. parvum, 2검체가 U. urealyticum이었으며불일치한 1검체는 U. parvum과 M. hominis 의복합감염으로동정된경우였다. Ureaplasma spp. 가 1 4 이하로배양된 41검체는모두가 PCR과일치하였고 U. parvum 38검체, U. urealyticum 3검체였다. M. hominis는 1검체에서배양되었는데 PCR 결과도동일하였다. U. parvum과 M. hominis이혼합되어자란 6검체중 5검체가일치하였으며 1검체에서는 PCR에서 U. parvum만양성이었다. 판독불가였던 1검체는 PCR에서 M. hominis로동정되었다. Mycoplasma IST2 kit (biomérieux) 를통해알아본항생제감수성검사결과는 Ureaplasma spp. 의감수성률이각각 clarithromycin 98.2% (56/57), erythromycin 86.% (49/57), azithromycin 8.7% (46/57) 였으며 doxycycline 94.7% (54/57), tetracycline 93.% (53/57) 그리고 ofloxacin 47.4% (27/57), ciprofloxacin 17.5% (1/57), josamycin 96.5% (55/57), pristinamycin % (57/57) 였다. 각각의균과그농도별로항생제감수성률 Table 1. Antimicrobial susceptibilities (%) of Ureaplasma spp. and M. hominis by Mycoplasma IST2 ERY AZI CLA JOS CIP OFL DOT TET PRI Isolates 1 4 < < < < < < < < <1 4 (38/38) (14/14) 92.1 (35/38) 92.9 (13/14) 92.1 (35/38) (14/14) 55.3 (21/38) 42.9 (6/14) 23.7* (9/38) 7.1* (1/14) (38/38) 85.7 (12/14) 97.4 (37/38) (14/14) 89.5* (34/38) 64.3* (9/14) 97.4* (37/38) Up 57.1* (8/14) (/3) (/2) (/3) (/2) (/2) Uu 5 (1/2) 66.7 (4/6) 16.7 (1/6) 16.7 (1/6) 16.7 (1/6) (/6) Mixed (Up+Mh) (/1) (/1) Mh (/1) *P<.5. Abbreviations: ERY, erythromycin; AZI, azithromycin; CLA, clarithromycin; JOS, josamycin; CIP, ciprofloxacin; OFL, ofloxacin; DOT, doxycycline; TET, tetracycline; PRI, pristinamycin; 1 4, titer 1 4 CFU in the specimen; <1 4, titer <1 4 CFU in the specimen; Up, U. parvum; Uu, U. urealyticum; Mh, M. hominis.

4 16 Ann Clin Microbiol 216;19(1):13-19 을살펴보면 Table 1과같았다. Erythromycin, azithromycin, ciprofloxacin의감수성률이균농도 1 4 이하보다 1 4 이상에서통계적으로유의하게낮았다 (P=., P=.1, P=.35) (Table 1). 분리된 M. hominis 균주는 erythromycin, azithromycin, clarithromycin 에는내성을나타냈으나 josamycin, pristinamycin, fluoroquinolones 그리고 tetracyclines에서는감수성을보였다. 2. U. parvum 과 U. urealyticum의 Macrolide, Tetracycline 과 Quinolone 내성유전자변이마크로라이드비감수성균주들에대한염기서열분석결과 23S rrna domain II와 L4와 L22 ribosomal proteins에서변이를보인중간내성균주의수는 azithromycin 3균주, erythromycin 4균주였고그변이종류는 U. urealyticum의경우 23S rrna domain II에서 I234V, F23L 을 L22 protein에서 A132V, I152T, I136V를보였고 U. parvum의경우 L22 protein에서 A121S, I136V, T141I를보였다. 그러나이들모두가감수성균주에서도관찰되어내성에는관여하지않는것으로생각되었다. 23S rrna domain V의염기서열분석에서는 double peaks가보여결과에포함시키지않았다 (Table 2). 테트라싸이클린비감수성균주들에대한 tet(m) 유전자와 int-tn PCR검사결과 tetracycline 내성균주 4검체중 3검체, doxycycline 내성균주 3검체중 2검체 doxycycline intermediate 균주 1검체에서 tet(m) 유전자또는 int-tn 유전자가발견되었으며이균주들모두 U. parvum이면서균농도는 1 4 이하였다 (Table 3). 퀴놀론비감수성균주들에대한 gyra,, parc, pare 유전자부위염기서열분석결과 ofloxacin 내성인 U. parvum 7균주중 5균주, ofloxacin 중간내성인 18균주중 2균주, ciprofloxacin 내성인 14균주중 7균주에서 quinolone 내성에관여하는 S83L 아미노산변이가관찰되었다. Quinolone 내성또는중간내성인 U. urealyticum 5균주중에서는내성에관여하는변이가관찰되지않았고, serovar에따라다른아미노산의차이만을보였다 (Table 4). Table 3. Presence of tet(m) or int-tn from tetracycline resistant, doxycycline non-susceptible isolates of Ureaplasma spp. Up<1 4 (n) DISCUSSION 상용화되어있는 Mycoplasma IST2 kit (biomérieux) 를사용하여배양된한국인여성의생식기마이코플라즈마의분리율은 44.% ( 균농도 1 4 이상 ; 11.3%, 1 4 이하 ; 27.3%) 로매우높게나타났으며이는생식기마이코플라즈마가한국인여성의생식기내집락화를이루고있음을보여주는결과였고과거에보여준 Koh 등 [17] 의연구결과와유사하였다. 그러나, U. parvum과 U. urealyticum의균종을감별한것은이연구가국내에서처음시행된것으로대부분이 U. parvum (8.3%) 으로동정되었고병원성이보고된 U. urealyticum은 7.6% 로관찰되었다. Mycoplasma IST2 kit (biomérieux) 과 PCR검사와의균검출의일치율은높았지만 Ureaplasma spp. 의감수성검사결과균의농도에따라 erythromycin, azithromycin, ciprofloxacin에서 TET-R (3) DOT-R (2) DOT-I (1) tet(m) 2 1 int-tn Abbreviations: TET, tetracycline; DOT, doxycycline; R, resistant; I, intermediate; <1 4, titer <1 4 CFU in the specimen; Up, U. parvum. Table 2. Amino acid changes in 23S rrna domain II, L4 and L22 ribosomal proteins from erythromycin intermediate, azithromycin intermediate isolates of Ureaplasma spp. Amino acid related to resistance Uu 1 4 < <1 4 Up ERY-I S rrna II I234V, F23L (1) L4 L22 A132V, I152T (1) A121S, I136V, T141I (2) A121S, T141I (1) AZI-I S rrna II F23L (1), I234V, F23L (1) L4 L22 A132V, I136V (1) A132V, I152T (1) A121S, T141I (1) Abbreviations: ERY, erythromycin; AZI, azithromycin; I, intermediate; 1 4, titer 1 4 CFU in the specimen; <1 4, titer <1 4 CFU in the specimen; Uu, U. urealyticum; Up, U. parvum.

5 Jiyoung Chang, et al. : Prevalence and Antimicrobial Susceptibility of Genital Mycoplasmataceae 17 Table 4. Amino acid changes in GyrA, GyrB, ParC and ParE from ofloxacin non-susceptible, ciprofloxacin non-susceptible isolates of Ureaplasma spp. Genes related to resistance Uu 1 4 < <1 4 Up OFL-R 3 4 gyra parc S83L (1) S83L (4) pare A429S (1), P446S (1) OFL-I gyra D112E (2) D112E (3) parc A125T, A136T (2) A125T, A136T (3) S83L (1) S83L (1) pare CIP-R gyra D112E (2) D112E (2) parc A125T, A136T (2) A125T, A136T (2) S83L (2) S83L (5) pare CIP-I gyra D112E (1) parc A125T, A136T (1) A136S (1) pare Abbreviations: OFL, ofloxacin; CIP, ciprofloxacin; R, resistant; I, intermediate; 1 4, titer 1 4 CFU in the specimen; <1 4, titer <1 4 CFU in the specimen; Uu, U. urealyticum; Up, U. parvum. 유의한차이를보여상품화된 kit의감수성검사의정확도가 inoculum의영향을받는것으로생각되었다. 하지만균농도를달리하여감수성시험을비교하는추가적인실험이필요할것으로생각된다. 또한, 동일계열의약제에대해서도감수성률이약제에따라차이가있어서 clarithromycin 98.2%, erythromycin 86.%, azithromycin 8.7% 순으로높았고퀴놀론계열의감수성률은 ofloxacin이 47.4% 로 ciprofloxacin (17.5%) 에비해높았다. 이러한차이는동일한검사방법으로시행한 Koh 등 [17] 의연구에서도항생제감수성률이 clarithromycin 88.6%, erythromycin 82.9%, azithromycin 75.2% 그리고 ofloxacin 56.2%, ciprofloxacin 15.2% 로유사하였다. 터어키의연구 [18] 에서는마크로라이드와퀴놀론계열모두 clarithromycin 88.9%, erythromycin 59.3%, azithromycin 55.6% 그리고 ofloxacin 11.1%, ciprofloxacin 7.4% 로더낮은감수성률을보였고, 중국에서의연구 [19] 와는마크로라이드계열은유사하였으나퀴놀론계열은 ofloxacin 22.1%, ciprofloxacin 5.8% 로더낮은감수성률을보였다. 이태리의연구 [2] 와는 clarithromycin 79.5%, erythromycin 69.1%, azithromycin 73.7% 그리고 ofloxacin 38.7%, ciprofloxacin 11.7% 로비교적유사하였지만, 스위스의연구 [21] 에서는퀴놀론계열이 ofloxacin 9.3%, ciprofloxacin 8.6% 로감수성률이높았다. Kit를이용하여살펴본표현형으로나타난내성양상과염기서열분석을통하여항생제내성유전자변이검사로본유전자형내성양상과의상호연관성을살펴본결과, 마크로라이드비감수성균주에서관찰된 23S rrna domain II와 L4와 L22 ribosomal protein 에서보이는모든아미노산변이가마크로라이드감수성균주에서도보여항균제내성과의연관성은없는것으로생각되었다. 테트라싸이클린내성균주에서는 75% (3/4) 에서내성유전자 tet(m) 과 int-tn을가지고있었다. 퀴놀론내성 U. urealyticum균주에서는 gyra D112E, parc A125T, A136T가관찰되었으나이들변이는 serovar에따른아미노산의차이로보고된바있는변이여서 [22] 내성과의연관성은없는것으로생각되었다. 이에비해퀴놀론내성 U. parvum에서는 parc 내성유전자변이로알려진 S83L 아미노산변이가 ofloxacin 내성균주에서는 7검체중 5균주 (71.4%) 에서 ciprofloxacin 내성균주에서는 14검체중 7균주 (5.%) 에서관찰되었으며, 1 4 이하이면서내성을보인경우에내성관련유전자변이가있는비율이더높은경향을보였다. Schneider 등 [21] 의연구에서는 Mycoplasma IST2 kit (biomérieux) 결과 ciprofloxacin 비감수성균주 13균주중 2균주만이 CLSI 지침에따른미세농도희석법으로시험했을때비감수성이었으며, 내성에관여하는유전자변이도 1균주에서만관찰되었다. 따라서, Mycoplasma IST2 kit (biomérieux) 를

6 18 Ann Clin Microbiol 216;19(1):13-19 이용한 quinolone 감수성결과의정확도에대해서는향후더많은검체로연구가필요하다. 또한, 2균주에서는 pare A429S, P446S 아미노산변이가관찰되었는데, 이는 serovar에따른차이는아니어서이변이의의미에대해서는추후연구가필요하다. 결론적으로 Mycoplasma IST2 kit (biomérieux) 를이용한생식기마이코플라즈마양성률은과거 29년진주에서시행한결과와유사한결과를보였고 [17] kit와 PCR검사간의일치율도높게나타났다. Ureaplasma spp. 가배양되는검체중대다수 (8.3%) 가 U. parvum이어서 U. urealyticum과 U. parvum을감별하여보고하는것이필요하며, 두가지균종이분리되는환자들의임상양상의차이에대한연구가필요하다. 또한, Ureaplasma 균종의 1% 이상에서 erythromycin과 azithromycin 에비감수성 (15.5%, 2.7%) 을보이므로이약제들을사용하기전감수성검사가필요할것으로생각된다. 염기서열분석을통한유전자변이와배양법에기반한항균제내성과의일치도는비교적낮았으나, 본연구에서는 CLSI guideline에따른감수성시험결과가아닌점과 efflux pump를비롯한다양한내성기전에대한유전자적검사는시행하지못한제한점이있다. 추후 ureaplasma의항생제내성에대한지속적인관심과연구가필요할것으로보인다. ACKNOWLEDGMENTS 본연구는 214년도대한임상미생물학회의연구비지원을받아수행되었음. REFERENCES 1. Waites KB, Katz B, Schelonka RL. Mycoplasmas and ureaplasmas as neonatal pathogens. Clin Microbiol Rev 25;18: Xiao L, Paralanov V, Glass JI, Duffy LB, Robertson JA, Cassell GH, et al. 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In vitro antimicrobial profile of Ureaplasma urealyticum from genital tract of childbearing-aged women in Nothern and Southern Italy. APMIS 214;122: Schneider SC, Tinguely R, Droz S, Hilty M, Donà V, Bodmer T, et al. Antibiotic susceptibility and sequence type distribution of Ureaplasma species isolated from genital samples in Switzerland. Antimicrob Agents Chemother 215;59: Beeton ML, Chalker VJ, Kotecha S, Spiller OB. Comparison of

7 Jiyoung Chang, et al. : Prevalence and Antimicrobial Susceptibility of Genital Mycoplasmataceae 19 full gyra,, parc and pare gene sequences between all Ureaplasma parvum and Ureaplasma urealyticum serovars to separate true fluoroquinolone antibiotic resistance mutations from non-resistance polymorphism. J Antimicrob Chemother 29;64: = 국문초록 = 한국인가임기여성에서 Ureaplasma Species 의빈도와항생제감수성 : 유전자변이와의일치 가톨릭대학교의과대학 1 진단검사의학교실, 2 산부인과학교실장지영 1, 유진경 1, 송창은 1, 박인양 2, 박연준 1 배경 : Mycoplasma IST2 kit (biomérieux, France) 로검출된생식기 mycoplasma 중 Ureaplasma urealyticum은 7.6% 관찰된반면대부분이 Ureaplasma parvum으로동정되었다 (8.3%). U. parvum과 U. urealyticum을감별한것은본연구가처음으로저자들은한국인여성의 ureaplasma 분리율과항생제내성을알아보고자하였다. 방법 : 214년 1월에서 11월까지서울성모병원에서 Mycoplasma IST2 kit (biomérieux) 를사용하여배양된 15검체를대상으로하였다. Mycoplasma IST2 kit로배양과항균제감수성검사를하였고 Ureaplasma spp. 동정을위하여 PCR 검사를, 내성유전자변이를보기위하여 PCR과 sequencing을시행하였다. 결과 : 생식기 mycoplasma의양성률은 44.% (66/15) 였고그중 53 (8.4%) 검체가 U. parvum으로동정되었으며 5 (7.6%) 검체는 U. urealyticum, 2 (3.%) 검체는 Mycoplasma hominis, 6 (9.%) 검체는복합감염을보였다. Ureaplasma spp. 에대한항균제감수성률은 erythromycin, azithromycin, clarithromycin과 doxycycline에서각각 86.%, 8.7%, 98.2%, 94.7% 였고 ofloxacin, ciprofloxacin에서는각각 47.4%, 17.5% 였다. Ofloxacin 내성균주와 (5/7, 71.4%), ciprofloxacin 내성균주에서 (7/14, 5.%) ParC 부위의 S83L 아미노산변이가각각관찰되었다. 한균주의 M. hominis 는 erythromycin, azithromycin, clarithromycin에는내성을, josamycin, pristinamycin, quinolone, tetracycline에는감수성을각각보였다. 결론 : 한국인여성의생식기 mycoplasma의분리율은 44.% 였고대부분은 U. parvum이었다. Ureaplasma spp. 의 erythromycin 과 azithromycin의비감수성률이 1% 이상보이므로 (15.5%, 2.7%) 이들약제를사용하기전항균제감수성검사가필요할수있다. U. urealyticum과 U. parvum 감염으로인한임상적양상차이와내성기전에대한추후연구가필요하겠다. [Ann Clin Microbiol 216;19:13-19] 교신저자 : 박연준, 6591, 서울시서초구반포대로 222 가톨릭대학교서울성모병원진단검사의학과 Tel: , Fax: yjpk@catholic.ac.kr

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( )Kju225.hwp 비임균성비뇨생식기감염에서 의진단적효용성 Usefulness of the Mycofast Test (MYCOFAST Evolution 2) for the Diagnosis of Nongonococcal Genitourinary Infections Hang Ro Park, Yang Hyun Kim, Ho Jae Lee, Jea Sang Oh, Hyoung Jin