• Clinical and Experimental Pediatrics
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• 제60권5호
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• pp.151-157
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• 2017

Purpose: Macrolide resistance rate of Mycoplasma pneumoniae has rapidly increased in children. Studies on the clinical features between macrolide susceptible-M. pneumoniae (MSMP) and macrolide resistant-M. pneumoniae (MRMP) are lacking. The aim of this study was to identify the macrolide resistance rate of M. pneumoniae in Korean children with M. pneumoniae penupmonia in 2015 and compare manifestations between MSMP and MRMP. Methods: Among 122 children (0-18 years old) diagnosed with M. pneumoniae pneumonia, 95 children with the results of macrolide sensitivity test were included in this study. Clinical manifestations were acquired using retrospective medical records. Results: The macrolide resistant rate of M. pneumoniae was 87.2% (82 of 94 patients) in children with M. pneumoniae pneumonia. One patient showed a mixed type of wild type and A2063G mutation in 23S rRNA of M. pneumoniae. There were no significant differences in clinical, laboratory, and radiologic findings between the MSMP and MRMP groups at the first visit to our hospital. The time interval between initiation of macrolide and defervescence was significantly longer in the MRMP group ($4.9{\pm}3.3$ vs. $2.8{\pm}3.1days$, P=0.039). Conclusion: The macrolide resistant rate of M. pneumoniae is very high in children with M. pneumoniae pneumonia in Korea. The clinical manifestations of MRMP are similar to MSMP except for the defervescence period after administration of macrolide. Continuous monitoring of the occurrence and antimicrobial susceptibility of MRMP is required to control its spread and establish strategies for treating second-line antibiotic resistant M. pneumoniae infection.

• 대한임상검사과학회지
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• 제41권4호
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• pp.153-157
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• 2009

The production of extended-spectrum ${\beta}$-lactamases ($ESBL_S$) of the TEM or SHV type by bacterial pathogens is a major threat to the use of the clinically important expanded-spectrum cephalosporins. The characterization of the SHV ESBLs producing Klebsiella pneumoniae strains present in clinical isolates is time-consuming processes. We describe here in the development of a novel system, which consists of a real time PCR. We found 11 K. pneumoniae strains to be presumptive strains ESBLs producers by clinical and laboratory standards institute (CLSI) guidelines. The double disk synergy test showed 8 ESBL positive and conventional PCR showed 10 SHV ESBL positive, which were K. pneumoniae strains isolates. By real time PCR analysis, SHV gene in 11 of 11 strains were identified. When sequencing analysis was compared with real time PCR, both analysis were presented 99% similarity. In this study, we used a rapid, sensitive, and specific real-time PCR (RT-PCR) method for detection of the assay SHV ESBL producing K. pneumoniae strains in clinical isolates.

• Archives of Pharmacal Research
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• 제19권3호
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• pp.173-177
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• 1996

The Pneumococcus, Streptococcus pneumoniae, has an ample polysaccharide (PS) capsule that is highly antigenic and is the main virulence factor of the organism. The capsular PS is the source of PS vaccine. This investigation was undertaken to optimize the culture conditions for the production of capsular PS by type 4 pneumococcus. Among several culture media, brain heart infusion (BHI) and Casitone based medium were found to support luxuriant growth of pneumococcus type 4 at the same level. Therefore in this study, the Casitone based medium was used to study optimization of the culture condition because of BHI broth's high cost and complex nature. The phase of growth which accomodated maximum PS production was exponential phase. Concentrations of glucose greater than 0.8% did not enhance growth or PS production. Substitution of nitrogen sources with other resources or supplementation of various concentrations of metal ion (with the exception of calcium, copper, and magnesium ions) had adverse effects on growth and PS production. On the other hand, low level aeration and supplementation of 3 mg/l concentration of asparagine, phenylalanine, or threonine were beneficial for increased PS production. The synergistic effect of all the favorable conditions observed in pneumococcal growth assays provided a two-fold cumulative increase in capsular PS production.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2008년도 International Meeting of the Microbiological Society of Korea
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• pp.23-25
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• 2008

Serum complement proteins comprise an important system that is responsible for several innate and adaptive immune defence mechanisms. There were three well described pathways known to lead to the generation of a C3 convertase, which catalyses the proteolysis of complement component C3, and leads to the formation of C3 opsonins (C3b, iC3b and C3d) that fix to bacteria. A pivotal step in the complement pathway is the assembly of a C3 convertase, which digests the C3 complement component to form microbial-binding C3 fragments recognized by leukocytes. The spleen clears microorganisms from the blood. Individuals lacking this organ are more susceptible to Streptococcus pneumoniae. Innate resistance to S. pneumoniae has previously been shown to involve complement components C3 and C4, however this resistance has only a partial requirement for mediators of these three pathways, such as immunoglobulin, factor B and mannose-binding lectin. Therefore it was likely that spleen and complement system provide resistance against blood-borne S. pneumoniae infection through unknown mechanism. To better understand the mechanisms involved, we studied Specific intracellular adhesion molecule-grabbing nonintegrin (SIGN)-R1. SIGN-R1, is a C-type lectin that is expressed at high levels by spleen marginal-zone macrophages and lymph-node macrophages. SIGN-R1 has previously been shown to be the main receptor for bacterial dextrans, as well as for the capsular pneumococcal polysaccharide (CPS) of S. pneumoniae. We examined the specific role of this receptor in the activation of complement. Using a monoclonal antibody that selectively downregulates SIGN-R1 expression in vivo, we show that in response to S. pneumoniae or CPS, SIGN-R1 mediates the immediate proteolysis of C3 and fixation of C3 opsonins to S. pneumoniae or to marginal-zone macrophages that had taken up CPS. These data indicate that SIGN-R1 is largely responsible for the rapid C3 convertase formation induced by S. pneumoniae in the spleen of mice. Also, we found that SIGN-R1 directly binds C1q and that C3 fixation by SIGN-R1 requires C1q and C4 but not factor B or immunoglobulin. Traditionally C3 convertase can be formed by the classical C1q- and immunoglobulin-dependent pathway, the alternative factor-B-dependent pathway and the soluble mannose-binding lectin pathway. Furthermore Conditional SIGN-R1 knockout mice developed deficits in C3 catabolism when given S. pneumoniae or its capsular polysaccharide intravenously. There were marked reductions in proteolysis of serum C3, deposition of C3 on organisms within SIGN-$R1^+$ spleen macrophages, and formation of C3 ligands. The transmembrane lectin SIGN-R1 therefore contributes to innate resistance by an unusual C3 activation pathway. We propose that in the SIGN-R1 mediated complement activation pathway, after binding to polysaccharide, SIGN-R1 captures C1q. SIGN-R1 can then, in association with several other complement proteins including C4, lead to the formation of a C3 convertase and fixation of C3. Therefore, this new pathway for C3 fixation by SIGN-R1, which is unusual as it is a classical C1q-dependent pathway that does not require immuno globulin, contributes to innate immune resistance to certain encapsulated microorganisms.

• 한국미생물·생명공학회지
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• 제34권4호
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• pp.342-351
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• 2006

병원내 항생제 다제 내성을 일으키는 CTX-M형 ESBL을 생성하는 E. coli와 Klebsielia pneumoniae의 생성현황을 조사하고 이들 균주로 인한 감염증치료와 역학적 조사연구에 도움이 되고자 효소의 유전형을 규명하였다. 2005년 7월-12월에 부산에 소재하고 있는 2개의 종합병원에서 분리된 E. coli와 K. pneumoniae 각각 153주, 52주를 수집하였다. 그 중에서 ESBL을 생성 하는 균주를 검출하기 위해 Double disk synergy test를 시행하여서 E. coli 23주와 K. pneumoniae 13주를 분리하였다. 균주의 동정은 Vitek system GNI card(bioMerieux Vitek Inc., Hazelwood, Mo., U.S.A.)로 확인하였고, 항생제감수성시험은 disk diffusion method 와 agar dilution method를 사용하였다. 분리된 균주들의 내성을 일으키는 ESBL유전형을 규명하기 위하여 Isoelectric focusing(IEF), polymerase chain reaction test, DNA sequencing을 시행하였다. A병원의 13주와 B병원의 10주로 총 23주의 E. coli(15.0%)와 A병원의 7주와 B병원의 6주로 K. pneumoniae 13주(25.0%)가 double disk synergy test 양성으로 ESBL 생성균주로 판정하였다. ESBL 생성 36균주를 대상으로 bla$_{TEM}$, bla$_{SHV}$, bla$_{CTX-M}$ 유전자 검출을 위한 PCR을 시행한 결과 bla$_{TEM}$ 유전자는 13주(36.1%), bla$_{SHV}$ 유전자는 13주(36.1%), bla$_{CTX-M}$ 유전자는 32주(88.9%)가 양성반응을 보여서 bla$_{CTX-M}$ 유전자를 가진 균주가 가장 많이 나타났다. 그리고, bla$_{TME}$, bla$_{SHV}$ 두 가지 유전자를 가지고 있는 균주는 1주(2.8%)만 나타났고 bla$_{TEM}$, bla$_{CTX-M}$두 가지 유전자를 가지고 있는 균주는 9주(25.0%), bla$_{SHV}$, bla$_{CTX-M}$ 두 가지 유전자를 가지고 있는 균주가 10주(27.8%)로 나타나 bla$_{CTX-M}$을 포함하는 복합유전자가 많이 증가함을 알 수 있었다. 또한 CTX-M형 ESBL을 생성하는 E. coli와 K. pneumoniae에 대한 cefutaxime의 MIC는 256 $\mu$g/m1 이상으로 ceftazidime의 16-256 $\mu$g/mL 이상보다 높은 분포를 보였다. 즉, CTX-M형 ESBL 유전자를 지닌 균주에 대한 cefotaxim의 MIC는 ceftazidime의 MIC에 비해서 상대적으로 높은 양상을 보였다. 이러한 결과는 국내의 대학병원 뿐 만 아니라 일반종합병원에서도 CTX-M형 ESBL 생성 E. coli와 K. pneumoniae가 존재하며 확산 중임을 시사한다. 앞으로 CTX-M형 ESBL의 만연과 변종 CTX-M형 ESBL의 출연을 감시하기 위한 정기적인 연구와 조사가 필요한 것으로 생각한다.

• 미생물학회지
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• 제45권2호
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• pp.163-169
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• 2009

본 논문은 부산소재 하수처리장 중계시설인 민락오수처리장의 하수에서 광범위 베타 락탐 분해효소(extendedspectrum $\beta$-lactamase, ESBL) 생성균주를 분리 동정하고 이들이 생성한 ESBL 유형을 조사하였다. 민락오수처리장은 부산 남구 수영구의 민락동 일대에 밀집한 횟집의 하수를 모아서 남구 용호동에 있는 하수종말처리장으로 중계시키는 시설이다. 2009 년 1 월 하수 검체로부터 항균제 이중 디스크 확산 시험(double disk synergy test)에 양성반응을 나타낸 19 균주를 1차 시험균주로 선택하였다. 인돌생성 시험 methyl-red, Voges-Proskauer, Simmon's citrate, decarboxylase-dihydrolase 및 당 발효능 생화학 시험을 통하여 Klebsiella pneumoniae (3 균주)와 Escherichia coli (16 균주)가 동정되었다. 이 균들을 공여균주로, Escherichia coli J53 (sodium $azide^r$)를 피전달균주로 하여 접합시험을 실시하여 plasmid로 매개된 4 균주 접합체를 얻었다. 이들로부터 plasmid를 추출하여 PCR로 유전자 증폭을 시켜 유전자를 분석하고 등전점을 조사한 결과 Klebsiella pneumoniae에서 생성된 ESBL 유형은 모두 SHV-12 (3 균주)였고, Escherichia coli에서 생성된 ESBL 유형은 SHV-12/TEM-1 (1 균주)였다.

• Journal of Microbiology and Biotechnology
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• 제31권8호
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• pp.1109-1114
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• 2021

Chlamydia pneumoniae is a type of pathogenic gram-negative bacteria that causes various respiratory tract infections including asthma. Chlamydia species infect humans and cause respiratory infection by rupturing the lining of the respiratory which includes the throat, lungs and windpipe. Meanwhile, the function of interleukin-4 (IL-4) in Ch. pneumoniae respiratory infection and its association with the development of airway hyperresponsiveness (AHR) in adulthood and causing allergic airway disease (AAD) are not understood properly. We therefore investigated the role of IL-4 in respiratory infection and allergy caused by early life Chlamydia infection. In this study, Ch. pneumonia strain was propagated and cultured in HEp-2 cells according to standard protocol and infant C57BL/6 mice around 3-4 weeks old were infected to study the role of IL-4 in respiratory infection and allergy caused by early life Chlamydia infection. We observed that IL-4 is linked with Chlamydia respiratory infection and its absence lowers respiratory infection. IL-4R α2 is also responsible for controlling the IL-4 signaling pathway and averts the progression of infection and inflammation. Furthermore, the IL-4 signaling pathway also influences infection-induced AHR and aids in increasing AAD severity. STAT6 also promotes respiratory infection caused by Ch. pneumoniae and further enhanced its downstream process. Our study concluded that IL-4 is a potential target for preventing infection-induced AHR and severe asthma.

• 생명과학회지
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• 제20권5호
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• pp.676-682
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• 2010

본 연구는 임상검체만이 아닌 주변 생활환경에도 광범위 베타 락탐분해효소를 생성하는 균주(extended-spectrum $\beta$-lactamase, ESBL)가 존재하는지를 확인하고 만약 존재할 경우 그 균주를 분리하고 ESBL유형을 알아보기 위하여 실시되었다. 부산 광안리 하수처리 방류수에서 이중 디스크 확산 검사 결과 양성반응을 나타낸 29균주를 선별하였다. 이중 sodium azide에 내성을 가진 피전달 균주인 Escherichia coli J5에 교차접합이 이루어진 15균주를 대상으로 indole, methyl-red, Voges-Proskauer, Simmon's citrate 시험과 decarboxylase-dihydrolase 및 여러 종류의 당 발효 시험 등 생화학 검사를 실시한 결과 Klebsiella pneumoniae(13균주)와 Escherichia coli(2균주)가 동정되었다. 등전점, PCR, 유전자서열 분석을 실시하여 ESBL 유형을 결정하였다. Klebsiella pneumoniae의 ESBL 생성유형은 SHV-12(4균주)와 SHV-12/TEM-1(9균주)의 2종류로 구분되었고, Escherichia coli의 ESBL 생성유형은 TEM-1(2균주)로 판명되었다.

• 미생물학회지
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• 제46권1호
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• pp.38-45
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• 2010

본 연구는 부산 환경관리공단 수영사업소 산하 수영공공하수종말처리장 하수로부터 광범위 베타락탐 분해효소(ESBL, extended-spectrum ${\beta}$-lactamase)의 유형을 파악하기 위하여 이루어졌다. 수영공공하수처리시설은 부산광역시의 동북부 생활하수와 수세식변기의 오수가 표준활성슬러지공법으로 처리되며 일 생활하수 처리량은 총 550,000톤이다. 이중디스크 확산 검사, 제 3세대 세파계열 항균제에 대한 최소억제농도 시험을 통하여 14균주를 선별하였다. Indole, methyl-red, Voges-Proskauer, Simmon's citrate, decarboxylase-dihydrolase 시험과 당 발효 시험 등 생화학 검사를 통하여 동정한 결과 Escherichia coli (10균주), Klebsiella pneumoniae (4균주)가 동정되었다. 이를 전달균주로, sodium azide에 내성을 가진 피전달 균주인 E. coli J53에 교차접합을 시켜 11균주 (E. coli 9균주, K.pneumoniae 2균주)가 접합이 이루어졌다. 접합자중 2균주는 ESBL 유전자를 전달받지 못하였고 9균주는 부모로부터 ESBL 유전자를 전달받았다. 등전점, 유전자서열과 단백질서열 분석 등을 통하여 피전달균주인 E. coli J53에 전달된 유전자유형은 TEM형의 모형인 TEM-1과 SHV-12형으로 규명되었다.

• Journal of Microbiology and Biotechnology
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• 제21권7호
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• pp.734-738
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• 2011

Streptococcus pneumoniae is a major cause of invasive infection in young infants and older adults. There are currently 90 capsular serotypes identified and 23 serotypes (1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19F, 19A, 20, 22F, 23F, and 33F) are responsible for about 90% of invasive disease. Among the more than 90 different S. pneumoniae serotypes, serotype 19A is globally very prevalent. A simplified purification procedure including adjustment of cell lysate pH to 4.5, fractionation with 50. 80% ethanol, and dialysis rendered capsular polysaccharide (CPS) in a yield of $31.32{\pm}3.11$ mg from 1 l culture (75% recovery after lyses). The product contained only 69.6 ${\mu}g$ of protein (99.78% purity) and 0.8mg (sum of the precipitants from 50~60%, 60~70%, and 70~80%) of nucleic acid (97.45% purity). The purified CPS was conjugated with bovine serum albumin; the product size ranged from 100 to 180 kDa.