• Title/Summary/Keyword: Species detection

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Comparative Evaluation of the VITEK 2 System and Species-specific PCR Methods for the Detection of Vibrio Species Isolated from Shrimp (새우에서 분리된 Vibrio species 동정을 위한 VITEK 2 system방법과 species-specific PCR방법 비교 평가)

  • Lee, Jeong-Min;Lee, Won-Jun;Kim, Min-Ju;Cho, Yong-Sun;Lee, Jin-Sung;Lee, Hyun-Jin;Yoon, Sang-Woo;Kim, Keun-Sung
    • Journal of Food Hygiene and Safety
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    • v.30 no.3
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    • pp.281-288
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    • 2015
  • Vibrio is a genus of Gram-negative, curved, halophilic, and non-spore-forming bacteria. Some of the Vibrio species, such as V. cholerae and V. parahaemolyticus, often contaminate seafood products and occasionally cause human diseases when the seafood products are ingested. A total of 24 Vibrio strains were isolated from shrimp samples on Thiosulphate citrate bile salt sucrose (TCBS) media in this study. All of the 24 isolates were confirmed to belong to the genus Vibrio by using 16S rRNA gene sequence analyses. Vitek 2 system and species-specific polymerase chain reaction (PCR) methods were used to further identify a total of 29 Vibrio strains at the species level, including the 24 shrimp Vibrio isolates and five Vibrio reference strains. The specificities of the two methods to identify Vibrio strains at the species level were compared in this study. The species-specific PCR method was designed to detect five different Vibrio species, such as Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio alginolyticus, and Vibrio mimicus. From the 24 Vibrio shrimp isolates, the Vitek 2 system method could identify 15 (62.5%) strains as Vibrio species and 7 (29.2%) strains as non-Vibrio species, but could not identify the rest 2 (8.3%) strains. But species-specific PCR method could identify 16 (66.7%) strains as Vibrio species and could not identify the rest 8 (33.3%) strains. Among the 24 Vibrio shrimp strains, these two methods could unanimously identify 7 (7/24, 29.2%) strains (2 V. parahaemolyticus, 4 V. alginolyticus, and 1 V. mimicus). Considering that such different identification results were obtained using the two different methods in this study, identification method for Vibrio species must be carefully chosen.

Analysis of Healthcare Convergence on Bacterial Contamination of Radiological Equipment in Emergency Rooms of General Hospitals (종합병원 응급실내의 영상의학과 장비에서의 세균 오염에 관한 보건학적 융합 분석)

  • Hong, Dong-Hee;Park, Mi-Soon
    • Journal of the Korea Convergence Society
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    • v.8 no.1
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    • pp.51-59
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    • 2017
  • It is to establish preliminary data about healthcare convergencr of infection control by performing a bacterial contamination test for CT scanner as well as general radiographic systems in the emergency rooms of general hospitals. The period was December 1 ~ December 31, 2015. It is detect in the emergency rooms of 7 medical facilities including 2 in Seoul, and 1 in Gyeonggi-do Province, and 2 in Jeonla-do Province, and 2 in Chungcheong-do Province. The surface contamination strains detected from CT scanner in the radiology department emergency rooms were Micrococcus species(4,5%), Stenotrophomonas maltophilia(9%), Enterococcus faecium(4.5%), Providencia stuartii(4.5), and Gram negative bacilli(4.5%). Also, the surface contamination strains detected from general radiographic system in the radiology department emergency rooms were Providencia stuartii(11%), Klebsiella pneumonia(3.5%), Stenotrophomonas maltophilia(11%), Pantoea species(11%), Acinetobacter baumannii(3.5%), Micrococcus species(3.5%), Escherichia coli(3.5%), Enterobacter species(3.5%), and Gram negative bacilli(11%). Considering that the regions of most detection were all the places closely related to patients, radiologists would have to thoroughly clean with alcohol before and after a test.

The study on the Fluorescence Characteristics of Several Freshwater Bloom Forming Algal Species and Its Application (수종 담수적조 원인종들의 형광특성과 적용연구)

  • Son, Moon-Ho;Zulfugarov, Ismayil S.;Kwon, O-Seob;Moon, Byoung-Young;Chung, Ik-Kyo;Lee, Choon-Hwan;Lee, Jin-Ae
    • ALGAE
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    • v.20 no.2
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    • pp.113-120
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    • 2005
  • The freshwater blooms mainly blue-green algal blooms occur frequently in the lower Naktong River in summer, which provoke many socio-economical problems; therefore, the early detection of bloom events are demanding through the quantitative and qualitative analyses of blue green algal species. The in vivo fluorescence properties of cultured strains of Microcystis aeruginosa, M. viridis, M. wesenbergii, M. ichthyoblabe, Anabaena cylindrica, A. flos-aquae, and Synedra sp. were investigated. Wild phytoplankton communities of the lower Naktong River were also monitored at four stations in terms of their standing stocks, biomass and fluorescence properties compared with its absorption spectram. The 77K fluorescence emission spectra of each cultured strains normalized at 620 nm was very specific and enabled to detect of blue green algal biomass qualitatively and quantitatively. The relative chlorophyll a concentration determined by chlorophyll fluorescence analysis method showed significant relationship with chlorophyll a concentration determined by solvent extraction method ($R^2$ = 0.906), and the blue-green algal cell number determined by microscopic observation ($R^2$ = 0.588), which gives insight into applications to early detection of blue green algal bloom.

Molecular Detection of Ancylostoma duodenale, Ancylostoma ceylanicum, and Necator americanus in Humans in Northeastern and Southern Thailand

  • Phosuk, Issarapong;Intapan, Pewpan M.;Thanchomnang, Tongjit;Sanpool, Oranuch;Janwan, Penchom;Laummaunwai, Porntip;Aamnart, Witthaya;Morakote, Nimit;Maleewong, Wanchai
    • Parasites, Hosts and Diseases
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    • v.51 no.6
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    • pp.747-749
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    • 2013
  • The 2 principal species of hookworms infecting humans are Necator americanus and Ancylostoma duodenale. Case studies on zoonotic hookworm infections with Ancylostoma ceylanicum and/or Ancylostoma caninum are known mainly from Asian countries. Of these 2 zoonotic species, only A. ceylanicum can develop to adulthood in humans. In the present study, we report a molecular-based survey of human hookworm infections present in southern and northeastern Thailand. Thirty larval hookworm samples were obtained from fecal agar plate cultures of 10 patients in northeastren Thailand and 20 in southern Thailand. Partial ITS1, 5.8S, and ITS2 regions of the ribosomal DNA genes were amplified using PCR. The amplicons were sequenced, aligned, and compared with other hookworm sequences in GenBank database. The results showed that, in Thailand, N. americanus is more prevalent than Ancylostoma spp. and is found in both study areas. Sporadic cases of A. ceylanicum and A. duodenale infection were seen in northeastern Thailand.

Development of multiplex polymerase chain reaction for the detection of vancomycin resistant genotypes and Enterococcus Sp.-specific genes (장구균의 vancomycin 내성 유전자와 종 특이유전자의 검출을 위한 Multiplex polymerase chain reaction 개발)

  • Cho, Yun-Sang;Lee, Hee-Soo;Kim, Jong-Man;Ahn, Jong-Sam;Ryu, Pan-Dong;Park, Yong-Ho;Yoo, Han-Sang;Lee, Mun-Han
    • Korean Journal of Veterinary Research
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    • v.43 no.1
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    • pp.103-112
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    • 2003
  • A multiplex PCR assay, which allows simultaneous detection of vancomycin resistant genotypes and Enterococcus species-specific genes, was developed. Vancomycin resistant enterococci (VRE) from chickens and humans could be detected for vanA, vanB, vanC-1, vanC-2, $ddl_{E.faecium}$ and $ddl_{E.faecalis}$ by multiplex PCR. Eight isolates of VRE from humans (n=11) had $ddl_{E.faecium}$ and vanA, and 3 isolates of the VRE had $ddl_{E.faecium}$ and vanB. One isolate of VRE from chickens (n=6) had $ddl_{E.faecium}$ and vanA, and 5 isolates of the VRE had only vanA. E. faecium, E. faecalis, E. gallinarum and E. casseliflavus were also confirmed for the species-specific gene by multiplex PCR. This multiplex PCR could detect E. faecium, E. faecalis, E. gallinarum, E. casseliflavus, vanA, vanB, vanC-1 and vanC-2, simultaneously. The PCR assay established in the present study can be an alternative to time-consuming biochemical tests and antibiotic susceptibility tests of Enterococcus spp.

Development of the Large-Scale Oligonucleotide Chip for the Diagnosis of Plant Viruses and its Practical Use

  • Nam, Moon;Kim, Jeong-Seon;Lim, Seungmo;Park, Chung Youl;Kim, Jeong-Gyu;Choi, Hong-Soo;Lim, Hyoun-Sub;Moon, Jae Sun;Lee, Su-Heon
    • The Plant Pathology Journal
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    • v.30 no.1
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    • pp.51-57
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    • 2014
  • A large-scale oligonucleotide (LSON) chip was developed for the detection of the plant viruses with known genetic information. The LSON chip contains two sets of 3,978 probes for 538 species of targets including plant viruses, satellite RNAs and viroids. A hundred forty thousand probes, consisting of isolate-, species- and genus-specific probes respectively, are designed from 20,000 of independent nucleotide sequence of plant viruses. Based on the economic importance, the amount of genome information, and the number of strains and/or isolates, one to fifty-one probes for each target virus are selected and spotted on the chip. The standard and field samples for the analysis of the LSON chip have been prepared and tested by RT-PCR. The probe's specific and/or nonspecific reaction patterns by LSON chip allow us to diagnose the unidentified viruses. Thus, the LSON chip in this study could be highly useful for the detection of unexpected plant viruses, the monitoring of emerging viruses and the fluctuation of the population of major viruses in each plant.

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.

Bacterial Community Monitoring of Commercial Kimchi Produced in Korea and China with Evidence of Bacilli Spore Formation during Fermentation (한국산 및 중국산 김치의 Bacteria 군집 분석 및 발효과정 중 Bacilli 포자 형성 규명)

  • An, Doohyun;Kim, Hye-Rim;Jeong, Do-Won;Caldwell, Jane M.;Lee, Jong-Hoon
    • Microbiology and Biotechnology Letters
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    • v.42 no.2
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    • pp.121-130
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    • 2014
  • Terminal Restriction Fragment Length Polymorphism (T-RFLP) analysis was adopted to explore rapid differentiation in the diversity and dynamics of bacteria in kimchi made in Korea and China for future application in kimchi origin discrimination. T-RFLP analysis supported the reproducible and rapid detection of major lactic acid bacteria known to be involved in kimchi fermentation. The taxonomic resolution level of this T-RFLP analysis was between the species and genus level, but was not specific enough for the detection of a bacterium found only in one origin, either Korea or China. The bacterial community structure successions in kimchi samples from Korea and China analyzed by T-RFLP analysis occurred with a similar pattern. Bacillus spp. which were not detected in the early microbial studies of kimchi were constantly detected until the late fermentation stage of kimchi in our T-RFLP analysis and their existence was proved by culture-based identification. Additionally, sporulation of Bacillus spp. during kimchi fermentation was discovered.

Specific Detection of Root Rot Pathogen, Cylindrocarpon destructans, Using Nested PCR from Ginseng Seedlings (Nested PCR 기법을 이용한 인삼 뿌리썩음병원균의 특이적 검출)

  • Jang, Chang-Soon;Lee, Jung-Ju;Kim, Sun-Ick;Song, Jeong-Young;Yoo, Sung-Joon;Kim, Hong-Gi
    • Research in Plant Disease
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    • v.11 no.1
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    • pp.48-55
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    • 2005
  • Cylindrocarpon destructans is a soil-borne plant pathogenic fungus causing root rot on ginseng and trees. Rapid and exact detection of this pathogen was practiced on ginseng seedlings by nested PCR using speciesspecific primer set. The second round of PCR amplification by Dest 1 and Dest 4 primer set formed 400 bp of species-specific fragment of C. destructans from the product of first round of amplification by ITS 1 and ITS 4 primer set. In the PCR sensitivity test based on DNA density, nested PCR detected to the limit of one fg and it meant the nested PCR could detect up to a few spores of C. destructans. Also, nested PCR made it possible to detect the pathogen from ginseng seedlings infected by replantation on artificial infested soil. Our nested PCR results using species-specific primer set could be utilized for diagnosis of root rot disease in ginseng cultivation.

Evaluation of SERS Nanoparticles to Detect Bacillus cereus and Bacillus thuringiensis

  • Hong, Jeehwa;Qin, Jianwei;Van Kessel, Jo Ann S.;Oh, Mirae;Dhakal, Sagar;Lee, Hoonsoo;Hwang, Chansong;Chan, Diane E.;Kim, Dongho;Cho, Hyunjeong;Kim, Moon S.
    • Journal of Biosystems Engineering
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    • v.43 no.4
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    • pp.394-400
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    • 2018
  • Purpose: This research evaluated five types of nanoparticles to develop a surface-enhanced Raman spectroscopy (SERS) method for the rapid detection of two Bacillus species (Bacillus cereus and Bacillus thuringiensis) that are commonly found on fresh produce, which can cause food poisoning. Methods: Bacterial concentrations were adjusted to a constant turbidity, and a total of $30{\mu}L$ of each Bacillus cell suspension was prepared for each nanoparticle. A point-scan Raman system with laser light source of wavelength 785 nm was used to obtain SERS data. Results: There was no qualitative difference in the SERS data of B. cereus and B. thuringiensis for any of the five nanoparticles. Three gold nanoparticles, stabilized in either citrate buffer or ethanol, showed subtle differences in Raman intensities of two Bacillus species at $877.7cm^{-1}$. Conclusions: Among the three types of nanoparticles, the gold nanoparticles stabilized in citrate buffer showed the lowest standard deviation, followed by gold nanoparticles stabilized in ethanol. This result supports the potential application of gold nanoparticles for SERS-based detection of B. cereus and B. thuringiensis.