• 제목/요약/키워드: tryptic soy-serum-bacitracin-vancomycin medium

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Identification of Non-Aggregatibacter actinomycetemcomitans Bacteria Grown on the Tryptic soy-Serum-Bacitracin-Vancomycin Medium

  • Jo, Eojin;Park, Soon-Nang;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • 제41권4호
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    • pp.199-208
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    • 2016
  • The aim of this study was to identify the non-Aggregatibacter actinomycetemcomitans bacteria grown on the tryptic soy-serum-bacitracin-vancomycin (TSBV) medium, an A. actinomycetemcomitans selective medium. A total of 82 unidentified bacterial isolates from the oral cavities of a Korean population were kindly provide by the Korean Collection for Oral Microbiology. All the clinical isolates were grown on TSBV medium and bacterial DNA purified from each isolate was subjected to PCR with universal primers specific for bacterial 16S rRNA genes (16S rDNAs) sequence. The each bacterial 16S rDNA was amplified by PCR and the nucleotide sequences of it was determined by the dideoxynucleotide chain termination method. They were identified by 16S rDNA sequence comparison method at the specie-level. The data showed that Neisseria spp. (42 strains), Fusobacterium spp. (10 strains), Capnocytophaga spp. (8 strains), Propionibacterium acnes (5 strains), Aggregatibacter aprophilus (4 strains), Campylobacter spp. (5 strains), Veillonella dispar (3 strains), Streptococcus sp. (1 strain), Haemophilus parainfluenzae (1 strain), Leptotrichia wadei (1 strain), Morococcus sp./Neisseria sp. (1 strain), and Staphylococcus sp. (1 strain) were identified. These results could be used to develop a new A. actinomycetemcomitans-selective medium which is more effective than the TSBV medium in future studies.

한국인 Actinobacillus actinomycetemcomitans 균주의 특이 독성 clone형과 혈청형 및 백혈구독성과의 관계 (INTERRELATIONSHIP BETWEEN VIRULENT CLONAL TYPES, SEROTYPES AND LEUKOTOXICITY OF KOREAN STRAINS OF A. ACTINOMYCETEMCOMITANS)

  • 구영
    • Journal of Periodontal and Implant Science
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    • 제25권3호
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    • pp.487-496
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    • 1995
  • Previous studies have demonstrated that not all A. actinomycetemcomitans produced significant level of leukotoxic factor and its leukotoxicity have associated with serotype and genetic variation. Our aim was to investigate on the interrelationship between serotype and leukotoxicity of an A. actinomycetemcomitans consisting of 13 clinically well characterized. Korean isolates and to evaluate if particular virulent clonal types of A. actinomycetemcomitans are associated with periodontal disease. For this study, 13 strains of A. actinomycetemcomitans from 6 patients with periodontal disease were isolated and identified by using a selective medium(tryptic soy agar supplemented with 10% serum, $75{\mu}g$ of bacitracin and $5{\mu}g$ of vancomycin per ml) in 10% C02 incubator for 3days with routine Gram staining, colony morphology and biochemical test..For serotyping, antisera were prepared from reference strains of 5 serotypes. (ATCC 29523,Y4, SUNY aB 67, IDH 781, IDH 1705) and then ammonium sulfate precipitation, immunoabsorption and indirect immunofluoroscent procedures were done. For analysis of leukotoxicity, sonic extract of A. actinomycetemcomitans exposed to PMN, and trypan blue was stained for counting the cell viability. Finally Southern blot analyses of genomic DNA digested with the restriction enzyme Tag I was done and the Southern blots were hybridized with the 530bp fragment, termed delta 530, originating from the ltx promoter of strain 652 and deleted from strain JP2. Also ltxA-3.1 and SC2 probe from strain JP2 were hybridized with genomic DNA fragments. Results reveal that strains isolated showed approximately equal proportions of 3 serotypes(b, d, e) and serotype b was not detected. 2 patients harbored 2 different serotypes in the same disease site. The prevalence of leukotoxic strain was 23% and there was no relationship between serotype, leukotoxicity and clinical observations. Especially virulent clonal types of Actinobacillus actinomycetemcomitan (JP2 strain) could not found. Further studies are necessary on the genetic polymorphism of leukotoxin and its relations to clinical status.

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한국인 국소 유년성 치주염환자의 Actinobacillus(Haemophilus) Actinomycetemcomitans 혈청형 및 백혈구독성 균주 분포 (Serotype and Leukotoxic Strain Distribution of Actinobacillus(Haemophilus) Actinomycetemcomitans in Korean Localized Juvenile Periodontitis)

  • 정현주;정종평;손성희
    • 대한미생물학회지
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    • 제21권4호
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    • pp.487-501
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    • 1986
  • 국소적 유년성 치주염의 원인균으로 중요시되고 있는 Actinobacillus Actinomycetemcomitans(Aa)는 구미인의 병소에서 혈청형 b형이 주종을 이루는 것으로 보고되었으나, 한국인에서의 부분적인 분리균주는 c형이 빈번한 젓으로 관찰되었다. 이에 본 연구는 16명의 국소적 유년성 치주염 환자에서 Aa의 발현빈도를 조사하고 혈청형별로 분류하여 그 분포 및 백혈구 독성을 평가하기 위하여 시행되었다. 치주낭 및 건강치은 열구에서 보존치료용 paper point를 이용하여 치은연하 치태세균을 채취하여 Aa의 선택배지에 도말한 후 10% 탄산가스 배양기에서 $3{\sim}5$일간 배양하였으며, 집락의 형태, catalase검사, Gram염색,생화학검사로써 분리 동정하였다. 가토에서 3가지 혈청형의 표준균주인 ATCC 29523(a) Y4(b) SUNYaB67(c)에 대한 항혈청을 얻은 후 환산암모늄 침전법과 면역흡착법에 의하여 특이성을 갖는 감마글로블린액을 얻어서 ELISA법에 의해 분리균주의 혈청형을 분류하였다. 백혈구 독성은 다형핵 백혈구와 Aa분리균주를 함께 배양한 후 상층액에 대한 lactate dehydrogenase의 활성을 측정함으로써 평가하였다. 그 결과는 다음과 같다. 1. Aa균은 국소적 유년성 치주염 환자 16명중 75%에서 발견되었으며, 병소부위의 71%, 그리고 정상부위의 6%에서 나타났다. 2. 국소적 유년성 치주염 병소의 임상적 양상은 병소내 Aa의 존재여부에 따른 차이를 보이지 않았다. 3. 3가지 혈청형의 환자별 분포는 9명의 환자에서 유사하게 관찰되었으며, 3명의 환자에서는 동일한 구강내 또는 동일한 치주낭에서 다른 2가지 혈청형이 함께 분리되었다. 4. 백혈구 독성검사를 시행한 45개 균주중 22%에서 독성을 나타냈으며, 채취부위의 69%에서 백혈구 독성균이 존재하였다. 또한, 동일한 병소에서 독성균과 비독성균이 함께 관찰되었다. 5. 백혈구 독성균의 분포는 3가지 혈청형간에 차이를 인정할 수 없었다.

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