• Title/Summary/Keyword: Virulence PCR

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DD1.5k, the Gene Preferentially Expressed in Bloodstream Isolates of Vancomycin-Resistant Enterococcus faecium

  • Kim, Seung-Han;Lee, Dong-Gun;Yoo, Jin-Hong;Park, Su-Mi;Park, Jung-Hyun;Shin, Wan-Shik;Lee, Kyungwon;Dongeun Yong;Lee, Wee-Gyo
    • Journal of Microbiology
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    • v.42 no.2
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    • pp.143-146
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    • 2004
  • Vancomycin-resistant Enterococcus faecium (VREFM) is becoming a threatening pathogen. We identified a gene called DD1.5K by differential display-PCR, which was preferentially expressed in the bloodstream isolates of VREFM. Due to its amino acid similarity to transfer complex protein, trsE, and tissue-specific expression, this gene may be involved in virulence of VREFM.

Investigation on antimicrobial resistance genes of Salmonella Schwarzengrund isolated from pigs (돼지유래 Salmonella Schwarzengrund의 약제내성 유전자에 관한 연구)

  • Lee, Woo-Won;Kim, Sang-Hyun;Lee, Seung-Mi;Lee, Gang-Rok;Lee, Gi-Heun;Kim, Yong-Hwan
    • Korean Journal of Veterinary Service
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    • v.35 no.1
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    • pp.1-8
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    • 2012
  • To detect the virulence genes (invA and spvC) and antimicrobial resistance genes, polymerase chain reaction (PCR) was carried out using total 67 strains of S. Schwarzengrund isolated from pigs. As results, invA was detected from all 67 strains of S. Schwarzengrund, however, spvC was not at all. All 12 strains with ampicillin resistance, 15 strains with chloramphenicol resistance, 9 strains with kanamycin resistance, 1 strain with sulfamethoxazole/trimethoprim resistance, and 66 (98.5%) of 67 strains with tetracycline resistance carried TEM (${\beta}$-lactamase $bla_{TEM}$), cmlA (nonenzymatic chloramphenicol resistance), aphA1-Iab (aminoglycoside phosphotransferase), sulII (dihydropteroate synthase), and tetA (class A tetracycline resistance), respectively. All 63 strains with streptomycin resistance carried 3 aminoglycoside resistance genes, including aadA (aminoglycoside adenyltransferase), strA, and strB (streptomycin phosphotransferase). With respect to prevalence of antibiotic resistance genes occurred in S. Schwarzengrund, genes for strB (46.0%); strA and strB (30.2%); aadA, strA, and strB (9.5%); strA (7.9%); aadA and strB (3.2%); and aadA (3.2%) were detected by PCR.

Molecular subtyping and antimicrobial susceptibility of Streptococcus dysgalactiae subspecies equisimilis isolates from clinically diseased pigs

  • Oh, Sang-Ik;Kim, Jong Wan;Kim, Jongho;So, Byungjae;Kim, Bumseok;Kim, Ha-Young
    • Journal of Veterinary Science
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    • v.21 no.4
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    • pp.57.1-57.11
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    • 2020
  • Background: Streptococcus dysgalactiae subspecies equisimilis (SDSE) acts as an etiological agent for lameness, neurological signs, and high mortality in pigs. Despite its importance in pig industries and zoonotic potential, little is known about the effects of this pathogen. Objectives: This study aimed to determine the molecular characteristics and antimicrobial resistance of SDSE strains isolated from diseased pigs. Methods: A total 11 SDSE isolates were obtained from diseased pigs. Bacterial identification, PCR for virulence genes, emm typing, and antimicrobial resistance genes, multilocus sequence typing, and antimicrobial susceptibility test were performed. Results: Nine isolates were from piglets, and 8 showed lameness, sudden death, or neurological signs. The isolates were PCR-positive for sla (100%), sagA (100%), and scpA (45.5%), and only 1 isolate amplified the emm gene (stL2764). Eight different sequence types were detected, categorized into 2 clonal complexes and 4 singletons. All the isolates in this study were included in a small cluster, which also contained other strains derived from humans and horses. The minimum inhibitory concentrations for the tested beta-lactams were low, while those for macrolides, tetracyclines, and fluoroquinolones were relatively high. PCR analysis of the macrolide and tetracycline resistance genes demonstrated that the isolates carried erm(B) (18.2%, n = 2), mef(A/E) (9.1%, n = 1), tet(M) (18.2%, n = 2), and tet(O) (90.2%, n = 10). Two isolates presented a mutation in parC, which is associated with fluoroquinolone resistance. Conclusion: This study provided insight into swine-derived SDSE, as it is related to veterinary medicine, and elucidated its zoonotic potential, in the context of molecular epidemiology and antimicrobial resistance in public health.

Establishment of PCR to detect Bacillus anthracis in the experimentally infected soil and mice (PCR 기법을 이용한 인공감염토양 및 감염동물 장기로 부터 Bacillus anthracis의 검출)

  • Lee, Ji-youn;Yoo, Han-sang;Kim, Jong-yeom
    • Korean Journal of Veterinary Research
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    • v.38 no.3
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    • pp.574-580
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    • 1998
  • Anthrax caused by Bacillus anthracis is one of the most important zoonotic diseases in the worldwide. To control and prevent the disease effectively, several methods such as development of a fast and specific diagnostic method and vaccine, education etc, have been carried out. However, it still has a problem in the control and prevention. To control, the most important method is the prevention of direct or indirect contact of the causative agent with susceptible host. Therefore, we developed a fast and specific detection method, polymerase chain reaction, of B anthracis from soil and infected animals because the organism could survive long time in the environment including soil due to formation of spore. With the method, virulence genes of B anthracis were successfully amplified from experimentally infected soil and mice. Up to $4.2{\times}10$ of the organisms per gram could be detected with the PCR method from experimentally infected soil. These results suggested that this PCR method could be effectively used not only to detect B anthracis in soil and infected animal but also to provide the information to prevent the disease.

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Various Enterotoxin and Other Virulence Factor Genes Widespread Among Bacillus cereus and Bacillus thuringiensis Strains

  • Kim, Min-Ju;Han, Jae-Kwang;Park, Jong-Su;Lee, Jin-Sung;Lee, Soon-Ho;Cho, Joon-Il;Kim, Keun-Sung
    • Journal of Microbiology and Biotechnology
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    • v.25 no.6
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    • pp.872-879
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    • 2015
  • Many strains of Bacillus cereus cause gastrointestinal diseases, and the closely related insect pathogen Bacillus thuringiensis has also been involved in outbreaks of diarrhea. The diarrheal diseases are attributed to enterotoxins. Sixteen reference strains of B. cereus and nine commercial and 12 reference strains of B. thuringiensis were screened by PCR for the presence of 10 enterotoxigenic genes (hblA, hblC, hblD, nheA, nheB, nheC, cytK, bceT, entFM, and entS), one emetogenic gene (ces), seven hemolytic genes (hlyA, hlyII, hlyIII, plcA, cerA, cerB, and cerO), and a pleiotropic transcriptional activator gene (plcR). These genes encode various enterotoxins and other virulence factors thought to play a role in infections of mammals. Amplicons were successfully generated from the strains of B. cereus and B. thuringiensis for each of these sequences, except the ces gene. Intriguingly, the majority of these B. cereus enterotoxin genes and other virulence factor genes appeared to be widespread among B. thuringiensis strains as well as B. cereus strains.

Characterization of the rcsA Gene from Pantoea sp. Strain PPE7 and Its Influence on Extracellular Polysaccharide Production and Virulence on Pleurotus eryngii

  • Kim, Min Keun;Lee, Sun Mi;Seuk, Su Won;Ryu, Jae San;Kim, Hee Dae;Kwon, Jin Hyeuk;Choi, Yong Jo;Yun, Han Dae
    • The Plant Pathology Journal
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    • v.33 no.3
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    • pp.276-287
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    • 2017
  • RcsA is a positive activator of extracellular polysaccharide (EPS) synthesis in the Enterobacteriaceae. The rcsA gene of the soft rot pathogen Pantoea sp. strain PPE7 in Pleurotus eryngii was cloned by PCR amplification, and its role in EPS synthesis and virulence was investigated. The RcsA protein contains 3 highly conserved domains, and the C-terminal end of the open reading frame shared significant amino acid homology to the helix-turn-helix DNA binding motif of bacterial activator proteins. The inactivation of rcsA by insertional mutagenesis created mutants that had decreased production of EPS compared to the wild-type strain and abolished the virulence of Pantoea sp. strain PPE7 in P. eryngii. The Pantoea sp. strain PPE7 rcsA gene was shown to strongly affect the formation of the disease symptoms of a mushroom pathogen and to act as the virulence factor to cause soft rot disease in P. eryngii.

Prevalence and Characterization of Diarrheagenic Escherichia coli Isolated from Raw Chicken and Chilled Chicken in Korea (국내 유통중인 생닭 및 닭가공품에서 병원성 대장균의 분리 및 특성)

  • Cho, Yong-Sun;Lee, Da-Yeon;Kim, Hee-Eon;Lee, Myung-Ki;Lee, Joo-Young
    • Journal of Food Hygiene and Safety
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    • v.32 no.2
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    • pp.129-134
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    • 2017
  • Diarrheagenic Escherichia coli is now recognized as an important cause of diarrhea, hemorrhagic colitis and hemolytic-uremic syndrome (HUS) worldwide. E. coli were isolated from 80 of 356 (22.5%) chicken and chilled chicken products in Korea. Fifteen virulence genes specific for pathogenic E. coli, including Shiga toxin-producing E. coli (STEC), enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC) and enteroaggregative E. coli (EAEC), were examined by multiplex PCR. STEC virulence markers were detected for eaeA (20.0%), escV (21.3%), stx1 (3.8%), ent (2.5%), EHEC-hly (1.3%), stx2 (1.3%), EAEC virulence marker (astA) was detected in 32.5%. ETEC and EIEC were not detected. STEC serotypes O152, O1, O116, O26, O25, O119 and O153 were found in chicken samples. This suggests the importance of diarrheagenic Escherichia coli control in raw chicken and chilled chicken food for food safety.

Detection of plcR-papR Genes by PCR in Identifying Enterotoxin Genes-Harboring Bacillus cereus Strains (장독소 유전자 함유 Bacillus cereus 확인을 위한 독소 전사 조절 유전자 plcR-papR의 PCR 검출법)

  • Yun, Suk-Hyun;Kim, Yong-Sang;So, Soon-Ku;Jeong, Do-Yeon;Hahn, Kum-Su;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.45 no.4
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    • pp.425-429
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    • 2009
  • Identification of virulent Bacillus cereus strains was examined by PCR using primers specific for the detection of plcR-papR, which encode regulatory proteins controlling the transcription of virulence factors in B. cereus. Total 96 strains of B. cereus that carried at least one of diarrheal toxin genes including hblACD, nheABC, and cytK showed all positive PCR products, while other 48 Bacillus strains that lacked the toxin genes were plcRpapR-negative. This PCR method targeting the plcR-papR genes appears to be simple and effective in identifying the enterotoxin genes-harboring B. cereus strains.

Application of the rpoS Gene for Species-Specific Detection of Vibrio vulnificus by Real-Time PCR

  • Kim, Dong-Gyun;Ahn, Sun-Hee;Kim, Lyoung-Hwa;Park, Kee-Jai;Hong, Yong-Ki;Kong, In-Soo
    • Journal of Microbiology and Biotechnology
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    • v.18 no.11
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    • pp.1841-1847
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    • 2008
  • Vibrio vulnificus is a causative agent of serious diseases in humans, resulting from the contact of wound with seawater or consumption of raw seafood. Several studies aimed at detecting V. vulnificus have targeted vvh as a representative virulence toxin gene belonging to the bacterium. In this study, we targeted the rpoS gene, a general stress regulator, to detect V. vulnificus. PCR specificity was identified by amplification of 8 V. vulnificus templates and by the loss of a PCR product with 36 non-V. vulnificus strains. The PCR assay had the 273-bp fragment and the sensitivity of 10 pg DNA from V. vulnificus. SYBR Green I-based real-time PCR assay targeting the rpoS gene showed a melting temperature of approximately $84^{\circ}C$ for the V. vulnificus strains. The minimum level of detection by real-time PCR was 2 pg of purified genomic DNA, or $10^3$ V. vulnificus cells from pure cultured broth and $10^3$ cells in 1 g of oyster tissue homogenates. These data indicate that real-time PCR is a sensitive, species-specific, and rapid method for detecting this bacterium, using the rpoS gene in pure cultures and in infected oyster tissues.