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Serratia marcescens 검출을 위한 PCR 기법 개발 및 돼지정액 유래균주에 대한 항생제 감수성 양상
정지아(Ji-A Jung),김애란(Aeran Kim),서병주(Byoung Joo Seo),정석찬(Suk Chan Jung),김인철(In Cheul Kim),정기화(Ki Hwa Chung),정병열(Byeong Yeal Jung) 한국생명과학회 2013 생명과학회지 Vol.23 No.9
돼지 원정액의 채취나 희석정액의 제조과정 중 세균오염이 많이 발생하는데, 이는 정자활력의 감소뿐 아니라 모돈의 수태율 저하 등을 유발한다. 특히 Serratia marcescens는 환경에 널리 존재하며 비위생적으로 제조된 정액에 많이 분리되고 있다. 본 연구에서는 S. marcescens의 신속한 동정을 위하여 PCR 기법을 개발하였으며, 국내 돼지정 액 유래 S. marcescens을 이용하여 최소생육억제농도(MIC) 등을 조사하고 유효 항생제를 선발하고자 하였다. 개발 PCR 기법은 S. marcescens에서만 306 bp의 특이 유전자 증폭산물을 형성하였으며, 기타 돼지정액에서 분리 보고된 균주나 Serratia 속균에서는 유전자 증폭 산물이 형성되지 않아 특이성이 인정되었다. PCR 기법의 민감도는 S. marcescens에서 추출된 DNA 50 pg/μl까지 검출이 가능하였다. 디스크확산법에 의한 국내 돼지정액 유래 S. marcescens의 항생제 감수성을 조사한 결과, gentamicin, ceftiofur, neomycin 등에서 80% 이상의 높은 감수성을 보였 다. 한편 ceftiofur, enrofloxacin, gentamicin, neomycin의 MIC90는 각각 8, 8, 8, 16 μg/ml로 나타났다. 따라서 개발된 PCR 기법은 S. marcescens를 동정하는 유용한 방법이며, gentamicin 등 선발된 항생제는 S. marcescens에 의한 정액 내 세균오염을 관리하기 위한 희석제용 항생제로 추천된다. During the collection of boar semen, bacterial contamination usually occurs. The contamination has deleterious effects both on semen quality and on sow fertility. The majority of contaminants are gram-negative bacteria, especially Serratia marcescens. In this study, we developed a PCR assay for the identification of S. marcescens targeting the luxS gene (GenBank no. EF164926). S. marcescens yielded a specific 306 bp PCR product. However, no amplification was observed in the other strains tested. The detection limit of PCR was 50 pg/μl of template DNA of S. marcescens. The antimicrobial susceptibility patterns of S. marcescens isolated from boar semen were tested using the disk diffusion method. Gentamicin, ceftiofur, florfenicol, and neomycin showed high sensitivity in this test. The minimum inhibitory concentration (MIC) was also determined by the broth microdilution method. The MIC90 values of ceftiofur, enrofloxacin, gentamicin, and neomycin were 8, 8, 8, and 16 μg/ml, respectively. These results indicate that PCR amplification of the luxS gene is a reliable and effective method for the identification of S. marcescens and that ceftiofur, enrofloxacin, gentamicin, and neomycin are effective semen extenders for controlling S. marcescens.
Seol, Aeran,Shim, Yoo Jin,Kim, Sung Woo,Kim, Seul Ki,Lee, Jung Ryeol,Jee, Byung Chul,Suh, Chang Suk,Kim, Seok Hyun The Korean Society for Reproductive Medicine 2020 Clinical and Experimental Reproductive Medicine Vol.47 No.2
Objective: The purpose of this study was to determine the effect of vaginal progesterone for luteal phase support (LPS) on the clinical pregnancy rate (CPR) in natural frozen embryo transfer (FET) cycles via a meta-analysis. Methods: We performed a meta-analysis of randomized controlled trials (RCTs) and retrospective studies that met our selection criteria. Four online databases (PubMed, Embase, Medline, and the Cochrane Library) were searched between January 2017 and May 2017. Studies were selected according to predefined inclusion criteria and meta-analyzed using R software version 2.14.2. The main outcome measure was CPR. Results: A total of 18 studies were reviewed and assessed for eligibility. One RCT (n = 435) and three retrospective studies (n = 3,033) met the selection criteria. In a meta-analysis of the selected studies, we found no significant difference in the CPR (odds ratio [OR], 0.96; 95% confidence interval [CI], 0.60-1.55) between the vaginal progesterone and control groups. An analysis of the two retrospective cohort studies that reported the live birth rate (LBR) following FET showed a significantly higher LBR in the vaginal progesterone group (OR, 1.72; 95% CI, 1.21-2.46). A subgroup meta-analysis of FET conducted 5 days after injection of human chorionic gonadotropin showed no significant differences between the two groups with regard to the CPR (OR, 1.18; 95% CI, 0.90-1.55) or miscarriage rate (OR, 0.73; 95% CI, 0.36-1.47). Conclusion: The results of this meta-analysis of the currently available literature suggest that LPS with vaginal progesterone in natural FET cycles does not improve the CPR.
Rayamajhi, Nabin,Jung, Byeong Yeal,Cha, Seung Bin,Shin, Min Kyung,Kim, Aeran,Kang, Min Su,Lee, Kang Mu,Yoo, Han Sang American Society for Microbiology 2010 Applied and environmental microbiology Vol.76 No.14
<B>ABSTRACT</B><P>Fifteen nonrepetitive ampicillin-resistant <I>Salmonella</I> spp. were identified among 91 <I>Salmonella</I> sp. isolates during nationwide surveillance of <I>Salmonella</I> in waste from 131 chicken farms during 2006 and 2007. Additional phenotyping and genetic characterization of these 15 isolates by using indicator cephalosporins demonstrated that resistance to ampicillin and reduced susceptibility to cefoxitin in three isolates was caused by TEM-1 and DHA-1 β-lactamases. Plasmid profiling and Southern blot analysis of these three DHA-1-positive <I>Salmonella</I> serovar Indiana isolates and previously reported unrelated clinical isolates of DHA-1-positive <I>Salmonella</I> serovar Montevideo, <I>Klebsiella pneumoniae</I>, and <I>Escherichia coli</I> from humans and swine indicated the involvement of the large-size plasmid. Restriction enzyme digestion of the plasmids from the transconjugants showed variable restriction patterns except for the two <I>S</I><I>almonella</I> serovar Indiana isolates identified in this study. To the best of our knowledge, this is the first report of the presence of the DHA-1 gene among <I>Salmonella</I> spp. of animal origin.</P>
Byeong Yeal Jung(정병열),Bum Soo Park(박범수),Ha-Young Kim(김하영),Jae Won Byun(변재원),Aeran Kim(김애란),Albert Byungyun Jeon(전병윤),In Cheul Kim(김인철),Ki Hwa Chung(정기화) 한국생명과학회 2012 생명과학회지 Vol.22 No.8
돼지 정액의 세균오염은 정자활력 감소나 수태율 저하 등을 유발하는데, 특히 Stenotrophomonas maltophilia는 자연환경에 널리 존재하여 비위생적으로 채취된 정액에 많이 오염될 뿐 아니라 사람에게도 병원성을 일으킨다. 본 연구에서는 S. maltophilia의 검사 시간 단축과 정확도를 높이고자 PCR 기법을 개발하였으며, 돼지 원정액에서 이들 세균의 오염율을 조사하고 유효 항생제를 선발하고자 하였다. 돼지 원정액에서 분리 빈도가 높은 18 균종을 대상으로 PCR을 적용한 결과, S. maltophilia에서만 445 bp의 특이 유전자 증폭산물이 확인되었다. 또한 순수 배양된 S. maltophilia는 1.5×10³ CFU/ml까지, 원정액에 혼합된 S. maltophilia에 대해서는 1.5×10⁴ CFU/ml까지 검출이 가능하였다. 2009년에 116건과 2011년 324건 등 총 440건의 원정액 중 26건(5.9%)에서 S. maltophilia가 분리되었으며, 여름과 가을철에 오염율이 상대적으로 높게 나타났다. 균분리법과 PCR간의 일치율은 98.9%, kappa value는 0.906이었으며, PCR의 민감도와 특이도는 각각 100%, 98.7%로 나타났다. 분리균주의 항생제 감수성 시험 결과, enrofloxacin (100%)과 florfenicol (92.3%)에 높은 감수성을 보였으나 amoxicillin/clavulanic acid, apramycin, ceftiofur, penicillin, spectinomycin에는 내성율이 높게 나타났다. 결론적으로 개발된 PCR기법은 민감도, 특이도, 일치율이 높아 균분리법을 보조하고 신속?정확하게 정액 내 오염세균을 확인할 수 있어 효율적인 정액관리가 가능할 것으로 기대된다. Bacteria are frequently contaminated during the collection and processing procedures of boar semen. Of the contaminants, Stenotrophomonas (S.) maltophilia is a Gram-negative bacterium that is widely distributed in a variety of habitats. Although PCR assays have been developed for the detection of S. maltophilia, they cross-react with some species of Xanthomonas. In this study, we designed a primer set for the detection of S. maltophilia in order to target the chiA (GenBank accession no. NC_010943) gene. The specific PCR products were amplified from S. maltophilia only, not from other tested strains that are frequently found in semen. The detection limit of the PCR was 1.5×10³ CFU/ml with pure-cultured S. maltophilia and 1.5×10⁴ CFU/ml with S. maltophilia spiked in semen. Twenty-six (5.9%) S. maltophilia were isolated from 440 semen samples. The PCR results exhibited 98.9% agreement with a comparison of S. maltophilia isolation. Also, the sensitivity and specificity of the PCR were 100% and 98.7%, respectively. In the antimicrobial susceptibility test, S. maltophilia isolates were highly susceptible to enrofloxacin and florfenicol, while the majority of them were resistant to amoxicillin/clavulanic acid, apramycin, ceftiofur, penicillin, and spectinomycin. These results indicated that the PCR using the chiA gene was proven to be reliable and effective for the detection of S. maltophilia with high levels of sensitivity and specificity.
Lee, Ji Hyun,Jung, Aeran,Park, Han Na,Lee, Changhee,Mandava, Suresh,Lim, Sung-jun,Lim, Byoung-bok,Park, Sung-Kwan,Lee, Jongkook,Kang, Hoil Elsevier 2018 Forensic Science International Vol.291 No.-
<P><B>Abstract</B></P> <P>Illicit psychoactive substances have threatened public health worldwide. An active metabolite of ADB-CHMINACA and MDMB-CHMINACA was identified for the first time in a powder-type product found in an airmail package. The structure of compound 1 was elucidated by a combination of gas chromatography-mass spectrometry (GC–MS), liquid chromatography-high resolution mass spectrometry (LC-HRMS), infrared (IR) spectroscopy, and nuclear magnetic resonance (NMR) spectroscopy. Compound 1 was proven to be an analogue of MDMB-CHMINACA, an indazole-based synthetic cannabinoid. The methyl ester group in MDMB-CHMINACA was replaced with a carboxylic acid group in compound 1. Compound 1 was determined as 2-[1-(cyclohexylmethyl)-1<I>H</I>-indazole-3-carboxamido]-3,3-dimethylbutanoic acid and named as DMBA-CHMINACA.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A new indazole-3-carboxamide class synthetic cannabinoid was identified. </LI> <LI> Compound 1 was named as DMBA-CHMINACA according to the EMCDDA guideline. </LI> <LI> DMBA-CHMINACA is a hydrolysed form of MDMB-CHMINACA. </LI> <LI> DMBA-CHMINACA is a potential CB1 and CB2 agonist. </LI> <LI> DMBA-CHMINACA can work as a synthetic precursor of several synthetic cannabinoids. </LI> </UL> </P>