2003~2004년 경남 마산 지역에서 분리된 A군 연쇄구균의 T 혈청형과 emm 유전자형의 계절별 분포

전호상,박화진,이희주,마상혁,차성호,Jeon, Ho-Sang,Park, Hwa-Jin,Lee, Hee-Joo,Ma, Sang-Hyuk,Cha, Sung-Ho 대한소아감염학회 2005 Pediatric Infection and Vaccine Vol.12 No.1

목 적 : 류마티스열이나 사구체신염과 같은 합병증의 집단발생 및 GAS 감염의 역학조사를 위해서는 감염균의 혈청형분석이 필수적이다. T 단백은 GAS의 역학적 상황을 이용하는데 유용하여 선별검사로 가장 많이 이용한다. 혈청형 검사의 단점을 보완하기 위해 최근 분자생물학적 방법을 이용하기 시작하였다. 중합효소연쇄반응으로 emm 유전자를 증폭하여 자동염기서열 분석기를 이용한 emm형별 분석이 최근에 보고가 되고 있다. 그래서 저자들은 emm 유전자를 이용한 자동염기서열 분석과 T 혈청학적 분석을 통하여 마산의 어느 한 센터에서 얻은 GAS의 계절별 분포에 대해서 알고자 하였다. 방 법 : 2003년 6월부터 2004년 2월까지 경남 마산시에 위치한 창원파티마병원에서 급성 인두염이나 성홍열 또는 봉소염이 의심되는 환아에게 인후배양을 시행하였고, 그 중 GAS로 동정된 100개의 균주를 분리하여 미국 미네소타대학의 WHO Collaborating Center on Group A Streptococci에 보내 T 항원형을 혈청학적 동정을 하였고 emm 유전자 분석을 하였다. 이 결과들을 다시 계절별과 질병별로 분류하였다. 결 과 : T단백 혈청형은 총 19종이 분리되었다. 계절별로 분류하면 여름철에는 빈도순으로 T4(27.5%), T1(17.6%), T6(13.7%), T12(13.7%)이 흔히 분리되었고, 겨울철에는 T4(28.3%), T12(15.2%), T12/B3264(8.7%)의 순서로 흔히 발생하였다. T4, T12는 계절별로 변화가 거의 없었다. 질환별로 분리하면 급성 인두염은 89례를 차지하였는데 T 혈청형을 빈도순으로 나열하면 계절별 분포에서와 마찬가지로 T4(26.7%), T12(14.0%), T1(12.8%), T6(11.6%) 순으로 전체 균주의 빈도순과 동일한 양상을 보였다. emm 유전자형은 총 15종류가 분류되었다. emm 1, emm 6, emm 9, emm 44는 겨울철에 많이 감소하거나 없어졌고, emm 3, emm 12, emm 89는 겨울철에 새로 생기거나 빈도가 비교적 많이 증가한 유전자형이었다. 급성 인두염에서 emm 유전자형을 빈도순으로 나열하면 계절별 분포에서와 마찬가지로 emm 4(26.7%), emm 22(18.6%), emm 1(12.8%), emm 6(11.6%)의 순서로 분리되었다. 결 론 : 2003년과 2004년에 걸쳐 조사한 T단백 혈청형과 emm 유전자형의 계절별 분포는 각각 19종과 15종으로 다양하게 분리되었다. 이 중 EM 내성과 관련이 있는 T 혈청형으로는 T12, T28, T4, T1, NT 등으로 알려져 있는데 이들의 비율이 전체의 56.7%를 차지하였다. 따라서 항생제 선택에 있어서 EM 내성률이 높다는 사실을 고려해야 한다. T 혈청형과 emm 유전자형의 분석이 GAS 질병의 역학과 병인을 연구하는데 도움이 되므로 앞으로 매년 감시를 해야하고, 특히 침습성 연쇄구균 감염으로부터 얻은 GAS을 T 혈청형 분석과 emm 유전자형 분석을 통해서 감시해야 하겠다. Purpose : The aim of this study is to know seasonal distribution of group A streptococci obtained from one center using emm genotyping and T serotyping in Masan from 2003 through 2004. Methods : Among children who visited the Changwon Fatima Hospital at Masan, Korea from June 2003 through February 2004, 100 patients who had clinical findings of acute pharyngitis, scarlet fever, and cellulitis were confirmed as GAS by culture, and were enrolled in our study. All obtained GAS were sent to the WHO Collaborative Center for Reference and Research on Streptococci, University of Minnesota, Minneapolis for T serotyping and emm genotyping. We classified these results again according to seasonal and disease's entities. Results : 19 different T serotypes was typed. T4(27.5%), T1(17.6%), T6(13.7%), and T12(13.7%) serotypes were relatively common in summer, while T4(28.3%), T12(15.2%), and T12/B3264(8.7%) were common in winter. T4 and T12 were persistent all year around. Distribution of T serotypes in 89 patients with pharyngotonsillitis were T4(26.7%), T12(14.0%), T1(12.8%), and T6(11.6%) in order of frequency. 15 different emm genotypes was typed. The number of emm 1, emm 6, emm 9, and emm 44 genotypes decreased or disappeared in winter, and the number of emm 3, emm 12, and emm 89 genotypes increased or reappeared in winter. Conclusion : Because T serotyping and emm genotyping are useful tools for evaluating epidemiology and pathogenesis of group A streptococci, we should monitor these strains every year, and should serotype and genotype GAS obtained from the invasive streptococcal infections.

혈액에서 분리된 A군 연쇄구균의 emm 유전자형과 외독소 생산

김선주,김의종 대한임상병리학회 1999 대한임상병리학회지 Vol.19 No.6

A군 연쇄구균의 T 항원형과 emm 유전자형 연관성

정현주,고은하,김선주,맹국영,강성하 대한임상미생물학회 2006 Annals of clinical microbiology Vol.9 No.1

배경:A군 연쇄구균(group A streptococci, GAS) 감염이나 보균자 역학조사를 위해 선별검사로 T 타이핑이 이용되고, 병독성 연구를 위해 M 타이핑이 이루어져 왔다. 그러나 M 항원형 검사가 일반 검사실에서는 시행하기 어렵고, 최근 PCR, 염기서열 분석이 보편화하면서 M 항원의 코딩 유전자인 emm 유전자형 검사가 가능해졌다. 본 연구에서는 T 항원과 emm 유전자형의 연관성을 살펴보았다. 방법: 2002년부터 2004년까지 진주 지역 초등학생에서 분리된 349균주와 인두염 소아 환자에서 분리된 122균주를 대상으로 하였다. 슬라이드 응집법으로 T항원형 검사를 시행하였고, PCR 및 염기서열 분석법으로 emm 유전자형을 조사하였다. 결과: T1/emm1, T3/emm3, T6/emm6, T12/emm12와 emm22, T25/emm75 및 T5/27/44/emm44/61은 90%이상의 높은 연관성을 관찰할 수 있었지만, T 2, 4, 7, 11, B3264 등은 다양한 emm 유전자형 종류를 보였다. 결론:일부 T 항원형은 특정 emm 유전자형과 밀접한 연관성이 있어서 emm 유전자형을 예측하는데 도움을 주었다.

소아과 의원을 방문한 급성인두염 환자의 인후배양과 emm 유전자형을 이용한 A군 연쇄구균의 역학조사

정현주,이남용,권오영,맹국영,김선주,Jung, Hyun Ju,Lee, Nam Yong,Kwon, Oh-Young,Maeng, Kook Young,Kim, Sunjoo 대한소아감염학회 2003 Pediatric Infection and Vaccine Vol.10 No.2

목 적 : 소아과 의원을 방문한 인두염 환자에서 A군 연쇄구균 분리율을 살펴보고자 하였다. T 항원형과 emm 유전자형을 동정하여 역학적 특성을 살펴보고, 같은 지역의 보균자 결과와 비교하고자 하였다. 방 법 : 2001년 11월부터 2002년 5월까지 경남 진주시에 위치한 한 소아과 의원에서 인두염 환자 246명(남자 123명, 여자 123명)을 대상으로 인후배양을 시행하였다. 슬라이드 응집법으로 T 항원형을 결정하였고, emm 유전자 증폭 후 염기서열 분석을 시행하여 emm 유전자형을 동정하였다. 결 과 : 인두염 환자 246명 중 130명(52.8%)에서 베타용혈성 연쇄구균이 분리되었으며, 그 중 96.1%가 A군이었다. 연령별로는 4세에서 7세가 70.4%로 가장 많았다. T12가 35.2%로 가장 흔하였고, T non-typeable 30.4%, T28 14.4% 순이었다. emm12(28.5%)가 가장 많이 동정되었고, emm75(18.7%), emm22(13.0%), emm2(12.2%) 및 emm8(8.1%) 순이었다. 결 론 : A군 연쇄구균 인두염은 약 50%에서 분리될 정도로 흔하므로, 개인 소아과 의원에서 급성인두염 환자에 대해 항생제 처방 전에 인후배양이 필요할 것으로 사료된다. T항원 및 emm 유전자형은 급성 인두염 환자와 보균자에서 비슷한 분포를 보였다. Purpose : Group A streptococci(GAS) was isolated from the patients with acute pharyngitis. Epidemiological studies using T typing and emm genotyping was performed for GAS and compared with the results of the carriers. Methods : Throat cultures were taken from 246 children(123 boys, 123 girls) from November, 2001 to May, 2002 who visited a pediatrician's office located in Jinju, Gyeongnam province. T types were identified with slide agglutination and emm genotypes were identified with DNA sequencing after amplification of emm genes. Results : One hundred thirty(52.8%) out of 246 children yielded beta-hemolytic streptococci, of which 96.1% were group A. Children from 4 to 7 years old comprised 70.4% of the GAS positive group. T12 were the most common(35.2%) and T non-typeable strains were the next(30.4%). emm12 was most frequent(28.5%), and emm75(18.7%), emm22(13.0%), emm2(12.2%), and emm8(8.1%) were relatively common. Conclusion : Since GAS is so highly prevalent in acute pharyngitis, indeed being half of the population, good clinical practice dictates the systematic employment of throat culture for acute pharyngitis before prescribing antibiotics in a pediatric setting. The distribution of the T antigens and emm genes showed similar pattern between the acute pharyngitis and the carriers.

Decline in Erythromycin Resistance in Group A Streptococci from Acute Pharyngitis due to Changes in the emm Genotypes Rather Than Restriction of Antibiotic Use

고은하,김선주 대한진단검사의학회 2010 Annals of Laboratory Medicine Vol.30 No.5

Background: Group A streptococcus (GAS) is the most common cause of bacterial pharyngitis in children. Antibiotic resistance rates and emm genotypes of GAS isolated from patients with acute pharyngitis were studied in 2009. Methods: Throat cultures were taken from 499 children with acute pharyngitis in Jinju, Korea, in 2008-2009. A total of 174 strains (34.9%) of GAS were isolated, and antimicrobial susceptibility testing was performed using the disk diffusion method. The phenotypes of macrolide resistance and macrolide resistance genes were determined. The emm genotypes were identified using PCR and sequencing. The data were compared with those acquired in 2002 in the same region. Data on the annual macrolide production were collected between 1999 and 2008. Results: The resistance rates of GAS to erythromycin, clindamycin, and tetracycline were 4.6%, 2.9%, and 2.3%, respectively. The constitutive resistance rate was 62.5% for the erm(B) gene and 37.5% for the M phenotype of the mef(A) gene. emm4 was most frequently detected (28.2%), followed by emm89 (20.1%). Most of the erythromycin resistant strains had the emm28 genotype. We noted a gradual increase in macrolide production during the study period. Conclusions: The erythromycin resistance rate of GAS isolated from children with acute pharyngitis was significantly lower in 2009 (4.6%) than in 2002 (44.8%). We observed a remarkable change in the distribution of emm genotypes during the 7-yr period. The significant decline in erythromycin resistance in 2009 might be associated with a prominent decrease in the resistant genotype emm12 (3.4% in 2009 vs. 28.0% in 2002) rather than restriction of macrolide use. (Korean J Lab Med 2010;30:485-90)

Molecular Epidemiological Features and Antibiotic Susceptibility Patterns of Streptococcus dysgalactiae subsp. equisimilis Isolates from Korea and Japan

김선주,변정현,박현웅,이재현,Haruno Yoshida,Akiyoshi Shibayama,Tomohiro Fujita,Yuzo Tsuyuki,Takashi Takahashi 대한진단검사의학회 2018 Annals of Laboratory Medicine Vol.38 No.3

Background: The molecular characterization of Streptococcus dysgalactiae subsp. equisimilis (SDSE) has not yet been performed in Korea. This study aimed to find the differences or similarities in the clinical features, molecular epidemiological findings, and antimicrobial resistance patterns of SDSE from two countries (Korea and Japan). Methods: SDSE isolates were collected from Korea (N=69) from 2012-2016 and Japan (N=71) from 2014-2016. Clinical characteristics, emm genotypes, and sequence types (STs) were compared. Microdilution tests were performed using different antimicrobials, and their resistance determinants were screened. Results: Median ages were 69 years in Korea and 76 years in Japan. The most common underlying diseases were diabetes and malignancy. Blood-derived isolates comprised 36.2% and 50.7% of Korean and Japanese isolates, respectively; mortality was not different between the two groups (5.8% vs 9.9%, P =0.53). Among Korean isolates with 20 different combined ST-emm types, ST127-stG245 (N=16), ST128-stG485 (N=10), and ST138- stG652 (N=8) were prevalent. Among Japanese isolates with 29 different combined types, ST17-stG6792 (N=11), ST29-stG485 (N=7), and ST205-stG6792 (N=6) were prevalent. Resistance rates to erythromycin, clindamycin, and minocycline were 34.8%, 17.4%, and 30.4% in Korea and 28.2%, 14.1%, and 21.4% in Japan, respectively. Conclusions: SDSE infections commonly occurred in elderly persons with underlying diseases. There was a significant difference in the distribution of ST-emm types between the two countries. Antimicrobial resistance rates were comparable with different frequencies of resistance determinants in each country.

Prevalence of Complement-Mediated Cell Lysis-like Gene (sicG) in Streptococcus dysgalactiae subsp. equisimilis Isolates From Japan (2014–2016)

Takashi Takahashi,Tomohiro Fujita,Akiyoshi Shibayama,Yuzo Tsuyuki,Haruno Yoshida 대한진단검사의학회 2017 Annals of Laboratory Medicine Vol.37 No.4

Background: Streptococcus dysgalactiae subsp. equisimilis (SDSE; a β-hemolytic streptococcus of human or animal origin) infections are emerging worldwide. We evaluated the clonal distribution of complement-mediated cell lysis-like gene (sicG) among SDSE isolates from three central prefectures of Japan. Methods: Group G/C β-hemolytic streptococci were collected from three institutions from April 2014 to March 2016. Fifty-five strains (52 from humans and three from animals) were identified as SDSE on the basis of 16S rRNA sequencing data.; they were obtained from 25 sterile (blood, joint fluid, and cerebrospinal fluid) and 30 non-sterile (skin-, respiratory tract-, and genitourinary tract-origin) samples. emm genotyping, multilocus sequence typing, sicG amplification/sequencing, and random amplified polymorphic DNA (RAPD) analysis of sicG-positive strains were performed. Results: sicG was detected in 30.9% of the isolates (16 human and one canine) and the genes from the 16 human samples (blood, 10; open pus, 3; sputum, 2; throat swab, 1) and one canine sample (open pus) showed the same sequence pattern. All sicG-harboring isolates belonged to clonal complex (CC) 17, and the most prevalent emm type was stG6792 (82.4%). There was a significant association between sicG presence and the development of skin/soft tissue infections. CC17 isolates with sicG could be divided into three subtypes by RAPD analysis. Conclusions: CC17 SDSE harboring sicG might have spread into three closely-related prefectures in central Japan during 2014–2016. Clonal analysis of isolates from other areas might be needed to monitor potentially virulent strains in humans and animals.