Genetic Variations of Candida glabrata Clinical Isolates from Korea using Multi-locus Sequence Typing

Min Ji Kang(강민지),Kyung Eun Lee(이경은),Hyunwoo Jin(진현우) 한국생명과학회 2020 생명과학회지 Vol.30 No.2

Candida albicans가 칸디다 혈증의 주요 원인 균으로 알려져 있지만, 최근에는 non-albicans Candida (NAC)에 의한 심각한 감염이 증가하고 있다. NAC 중에서 C. glabrata는 두 번째로 병원성을 나타내는 원인 균이지만 C. glabrata의 구조, 역학 등의 기본적인 연구는 거의 없는 실정이다. 따라서 본 연구에서는 다양한 유형의 임상 표본으로부터 분리된 총 102개의 C. glabrata균주로 multi-locus sequence typing (MLST)을 수행하였다. FKS, LEU2, NMT1, TRP1, UGP1 및 URA3을 포함한 6개의 하우스키핑 유전자를 증폭하여 염기서열을 분석하였다. 총 3,345개의 DNA 염기서열 중 49개의 가변 염기서열 부위가 발견되었으며, 그 결과 102개의 균주에서 총 12개의 상이한 서열 유형을 확인하였고, 알려지지 않은 서열 유형(USTs) 중 UST1이 가장 많이 나타났다. 이 연구의 결과는 국내 C. glabrata 항생제 처방에 도움이 될 것으로 사료되며, 한국에서 유행하는 C. glabrata에 대한 추가 연구를 위한 기본 역학자료로 사용될 것으로 기대한다. Although Candida albicans is the major fungal pathogen of candidemia, severe infections by non-albicans Candida (NAC) spp. have been increasing in recent years. Among NAC spp., C. glabrata has emerged as the second most common pathogen. However, few studies have been conducted to investigate its structure, epidemiology, and basic biology. In the present study, multi-locus sequence typing (MLST) was performed with a total of 102 C. glabrata clinical isolates that were isolated from various types of clinical specimen. For MLST, six housekeeping genes—FKS, LEU2, NMT1, TRP1, UGP1, and URA3—were amplified and sequenced. The results were analyzed using the C. glabrata database. Out of a total of 3,345 base-pair DNA sequences, 49 variable nucleotide sites were found, and the results showed that 12 different sequence types (ST) were identified from the 102 clinical isolates. The data also demonstrated that the undetermined ST1 was the most predominant ST in Korea. Further, seven undetermined STs (UST) containing UST2-8 were classified at specific loci. The data from this study may provide a fundamental database for further studies on C. glabrata, including its epidemiology and evolution. The data may also contribute to the development of novel antifungal agents and diagnostic tests.

Candida glabrata 감염에 의해 발생한 엉덩허리근 농양 1예

김미,이재갑,정창수,장주현,안병무,송원근 대한감염학회 2010 감염과 화학요법 Vol.42 No.1

Infections due to Candida species are becoming more frequent in several patient population and settings. The proportion of non-albicans Candida spp. causing candidemia has increased during the recent decades. Especially, fungaemia due to Candida glabrata has reduced susceptibility to azoles. We report a case of iliopsoas abscess caused by Candida glabrata. A 51-year-old male diabetic patient was admitted with fever and both hip joint pain. Abdominal CT scan revealed huge left iliopsoas intramuscular abscess and left perinephric abscess. The abscess was drained percutaneously. Cultures of the pus were positive for Candida glabrata. The patient’s condition improved after abscess drainage and was discharged with oral antifungal agent.

Candida glabrata 감염에 의해 발생한 엉덩허리근 농양 1예

김미,정창수,장주현,안병무,송원근,이재갑 대한감염학회 2010 Infection and Chemotherapy Vol.42 No.1

Infections due to Candida species are becoming more frequent in several patient population and settings. The proportion of non-albicans Candida spp. causing candidemia has increased during the recent decades. Especially, fungaemia due to Candida glabrata has reduced susceptibility to azoles. We report a case of iliopsoas abscess caused by Candida glabrata. A 51-year-old male diabetic patient was admitted with fever and both hip joint pain. Abdominal CT scan revealed huge left iliopsoas intramuscular abscess and left perinephric abscess. The abscess was drained percutaneously. Cultures of the pus were positive for Candida glabrata. The patient’s condition improved after abscess drainage and was discharged with oral antifungal agent.

Probe-based qPCR Assay for Rapid Detection of Predominant Candida glabrata Sequence Type in Korea

Jinyoung Bae,Kyung Eun Lee,Hyunwoo Jin 대한의생명과학회 2019 Biomedical Science Letters Vol.25 No.4

Recent years have seen an increase in the incidence of candidiasis caused by non-albicans Candida (NAC) species. In fact, C. glabrata is now second only to C. albicans as the most common cause of invasive candidiasis. Therefore, the rapid genotyping specifically for C. glabrata is required for early diagnosis and treatment of candidiasis. A number of genotyping assays have been developed to differentiate C. glabrata sequence types (STs), but they have several limitations. In the previous study, multi-locus sequence typing (MLST) has performed with a total of 101 C. glabrata clinical isolates to analyze the prevalent C. glabrata STs in Korea. A total of 11 different C. glabrata STs were identified and, among them, ST-138 was the most commonly classified. Thus, a novel probe-based quantitative PCR (qPCR) assay was developed and evaluated for rapid and accurate identification of the predominant C. glabrata ST-138 in Korea. Two primer pairs and hybridization probe sets were designed for the amplification of internal transcribed spacer 1 (ITS1) region and TRP1 gene. Analytical sensitivity of the probe-based qPCR assay was 100 ng to 10 pg and 100 ng to 100 pg (per 1 μL), which target ITS1 region and TRP1 gene, respectively. This assay did not react with any other Candida species and bacteria except C. glabrata. Of the 101 clinical isolates, 99 cases (98%) were concordant with MLST results. This novel probebased qPCR assay proved to be rapid, sensitive, highly specific, reproducible, and cost-effective than other genotyping assay for C. glabrata ST-138 identification.

칸디다균에 감염된 침습성 진균 부비동염으로 인한 안와첨증후군

김수진,오동은 대한안과학회 2024 대한안과학회지 Vol.65 No.8

목적: 칸디다균에 감염된 침습성 진균 부비동염으로 인한 안와첨증후군을 경험하여 보고하고자 한다. 증례요약: 76세 남성이 3주 전 발생한 좌안 시력저하 및 좌안, 측두부에 이르는 통증을 주소로 내원하였다. 당뇨, 신부전과 협심증을 앓고 있었으며, 1개월 전 COVID-19 확진으로 입원 치료한 이력이 있었다. 조영제를 사용한 안와 전산화 단층 촬영 검사에서 양측 부비동염 및 좌측 시신경을 침범하는 조영증강 병변이 관찰되어 이비인후과에서 내시경 부비동수술을 시행하였고 Gömöri methenamine silver로 염색한 병리조직검사에서 칸디다균 소견, 균배양 검사에서 Candida glabrata 소견을 확인하였다. 이에 amphotericin B 정맥주사를 시행하였고, 이어서 itraconazole 경구약을 투약하였다. 치료 6개월 후 호전되어 퇴원했으나, 안와첨증후군으로 인한 좌안 광각무 및 전체 눈근육마비 소견은 남아있었다. 결론: 칸디다 감염으로 인한 침습성 진균 부비동염은 국제적으로 매우 드물게 보고되었으며, 특히 국내에서는 처음 보고되었다. 염증성 병변이 발생한 경우 안과 파급 가능성을 염두에 두고 균주 특성에 따른 평가 및 치료가 필요하다. Purpose: To report a case of orbital apex syndrome resulting from invasive fungal sinusitis attributed to the unusual Candida glabrata species. Case summary: A 76-year-old patient presented with persistent left-sided orbital pain, headaches, and blurred vision for 3 weeks. The patient had a history of uncontrolled diabetes, chronic kidney failure, and angina, and was treated for a COVID-19 infection 1 month prior to admission. Contrast-enhanced computed tomography scans revealed extensive bilateral sinusitis affecting the left optic nerve. The patient underwent endoscopic sinus surgery, which included nasal cavity biopsy, conducted by an otorhinolaryngologist. Histopathological analysis using Gömöri methenamine silver stains identified Candida. Cultures from sinus specimens later grew Candida glabrata. Treatment involved daily IV amphotericin B followed by oral itraconazole. After 6 months, the patient fully recovered from fungal sinusitis but continued to experience negative light perception in the left eye and progressed to complete ophthalmoplegia, consistent with orbital apex syndrome. Conclusions: Reports of invasive fungal sinusitis caused by Candida infections are rare globally. This report marks the first occurrence in this country. Effective management requires careful assessment and appropriate treatment tailored to the specific species, especially considering the risk of orbital extension of the inflammatory lesion.

Real-time PCR법을 이용한 Candida glabrata의 Azole 내성 유전자 발현 분석:Northern 교잡법과의 비교

채명종,송정원,신종희,이진솔,김수현,신명근,서순팔,양동욱 대한의진균학회 2008 대한의진균학회지 Vol.13 No.2

Background: One of main mechanisms responsible for acquired azole resistance of Candida glabrata is the increased drug efflux mediated ABS transporters, which are encoded by CgCDR1 and CgCDR2 genes. Objectives: We compared real-time reverse transcriptase PCR (RT-PCR) with northern hybridization for quantitative analysis of CgCDR1 and CgCDR2 expression in bloodstream isolates of C. glabrata. Methods: Nineteen blood isolates of C. glabrata were selected, including nine fluconazole susceptible (MIC ≤8 μg/ml), nine susceptible dose-dependent (S-DD, MIC 16~32 μg/ml), and one resistant (MIC 128 μg/ml), isolates. The expression of CgCDR1 and CgCDR2 was quantified using real-time RT-PCR with ROTOR Gene 3000 (Corbettet research, Austria). The results were compared with northern hybridization with sequence-specific probes. Results: Correlation of quantification results between real-time RT-PCR and northern hybridization yielded correlation coefficients of 0.92 for CgCDR1 and 0.82 for CgCDR2 gene. By both methods, no significant differences were observed in the levels of expression of CgCDR1 and CgCDR2 between fluconazole-susceptible isolates and S-DD isolates. In contrast, a strain with high fluconazole resistance (MIC 128 μg/ml) revealed a greater abundance of CgCDR1 by both methods, compared to the other isolates. Conclusion: This study show that real-time PCR method for C. glabrata RNA quantification correlates well with traditional northern hybridization and can be a valuable alternative to northern hybridization for rapid quantification of CgCDR1 and CgCDR2 genes in clinical isolates of C. glabrata.

Candida Glabrata에 의한 이하선 농양 1예

이승균,박용수,전은주,김규백 대한이비인후과학회 2004 대한이비인후과학회지 두경부외과학 Vol.47 No.10

The parotid gland is the most commonly infected salivary gland. Parotid absces ocurs mostly in elderly persons who are Staphylococcus aureus and anaerobic bacteria are mostly cultured in aspirates of pus from parotid abscess. We report a case of a parotid abscess that grew Candida glabrata on culture after fine-nedle aspiration biopsy. This organism has not been previously associated with this condition.

Impact of high MIC of fluconazole on outcomes of <i>Candida glabrata</i> bloodstream infection: a retrospective multicenter cohort study

Ko, Jae-Hoon,Peck, Kyong Ran,Jung, Dong Sik,Lee, Ji Yeon,Kim, Hyun Ah,Ryu, Seong Yeol,Jung, Sook-In,Joo, Eun-Jeong,Cheon, Shinhye,Kim, Yeon-Sook,Kim, Shin-Woo,Cho, Sun Young,Ha, Young Eun,Kang, Cheol- Elsevier 2018 Diagnostic microbiology and infectious disease Vol.92 No.2

<P><B>Abstract</B></P> <P>To evaluate the impacts of fluconazole minimum inhibitory concentration (MIC) according to primary antifungal agents on <I>Candida glabrata</I> bloodstream infection (BSI), a multicenter retrospective cohort study was conducted in Korea, concerning the time period from January 2010 to February 2016. A total of 197 adult patients with <I>C. glabrata</I> BSI were included in the study, and neutropenia (<I>P</I> = 0.026), APACHE II score (<I>P</I> = 0.004), and fluconazole resistance (HR 3.960, 95% CI 1.395-11.246, <I>P</I> = 0.010) were associated with 30-day mortality in multivariate analysis. In subgroup analysis, fluconazole MIC = 32 μg/mL in the azole-treated group (HR 6.691, 95% CI 1.569-28.542, <I>P</I> = 0.010) and fluconazole MIC ≥ 64 μg/mL in the non-azole-treated group (HR 3.337, 95% CI 1.183-9.411, <I>P</I> = 0.023) showed the highest hazard ratio (HR) for 30-day mortality. Increased fluconazole MIC was associated with poor outcome both in azole- and non-azole-treated patients with <I>C. glabrata</I> BSI.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A multicenter cohort study was conducted for <I>C. glabrata</I> BSI. </LI> <LI> Neutropenia, APACHE II, and fluconazole resistance were associated with mortality. </LI> <LI> Fluconazole resistance was associated with mortality even in non-azole treated group. </LI> <LI> Types of antifungal agents were not associated with outcome. </LI> </UL> </P>

Prevalence of yeast‐like fungi and evaluation of several virulence factors from feral pigeons in Seoul, Korea

Jang, Y.H.,Lee, S.J.,Lee, J.H.,Chae, H.S.,Kim, S.H.,Choe, N.H. Blackwell Publishing Ltd 2011 Letters in applied microbiology Vol.52 No.4

<P><B>Abstract</B></P><P><B>Aims: </B> Studies of pigeon‐borne yeasts have tended to focus on species, such as <I>Cryptococcus neoformans</I> and <I>Candida albicans</I>, with scant attention to feral pigeons in Korea. We studied the prevalence of yeasts from faecal samples of feral pigeons obtained in various public places in Seoul, Korea, and assessed their potential capacity as human pathogens.</P><P><B>Methods and Results: </B> Three hundred and six pigeon faeces samples were collected at city squares and parks in 21 localities in Seoul and Seoul Grand Park and analysed for yeast with conventional methods. Of the 306 samples, 126 (41·2%) were positive for yeast. Seventeen species of yeast were identified. The most frequent species were <I>Candida glabrata</I> (34·1%), <I>Candida famata</I> (12·7%), <I>Cryptococcus albidus</I> (14·3%) and <I>Cryptococcus laurentii</I> (7·9%). The yeast isolates were tested for virulence. Of the 116 isolates (ten isolates missing), 70·7% (<I>n</I> = 82) grew at 37°C. All the <I>Cryptococcus</I> spp. isolates possessed a capsule, 16·4% (<I>n</I> = 19) produced melanin, and 33·6% (<I>n</I> = 39) produced proteinase. Two <I>Ca.?glabrata</I>, a <I>Ca.?famata</I> and <I>Ca.?albicans</I> as well as three <I>C.?neoformans</I>, a <I>C.?laurentii</I> and <I>Ca.?albicans</I> isolates had three virulence factors. Accordingly, 29·3% (<I>n</I> = 34) isolates possessed more than two virulence factors except capsule formation.</P><P><B>Conclusions: </B> These results of this study indicate that feral pigeons harbour a variety of yeasts and are a reservoir of human pathogenic fungi.</P><P><B>Significance and Impact of the Study: </B> This study is the first time about the microflora (fungi) presents in faecal samples collected from a variety of public areas throughout Seoul, Korea.</P>

Isolation of Green-Tea Polyphenols and Their Anticandidal Effects

이주희,한용문 동덕여자대학교 종합약학연구소 2002 동덕약학연구지 Vol.6 No.-

In a search of new anticandidal agents, tea polyphenolic compound was examined against Candida albicans and C. glabrata. The compound was extracted in hot water-ethyl acetate. Thin layer chromatography (TLC) analyses revealed that the extract (DTP) contained polyphenolic compound including EGCG as compared with commercially available polyphenols (TP) and epigallocatechin gallate (EGCG). An agar diffusion susceptibility assay showed that the DPT had growth-inhibitory effect on C. glabrata, whereas it did not inhibit C. albicans growth. The anti-C. glabrta effect of DTP at 1.25㎎/㎖ appeared to be equivalent to amphotericin B at 50 ㎍/㎖. However, this growth-inhibition was maintained only for 24 hrs and there was a partial growth-recovery, which was not observed with amphotericin B. Thus, it is considered that tea polyphenols may have fungistatic effect rather than fungicidal one.