• Title/Summary/Keyword: Bacterial

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Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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Measurement of the Levels of IgG Subclasses Reactive to Salmonella typhi in the Sera of Patients with Typhoid Fever (장티푸스환자의 혈청내 Salmonella typhi에 대한 IgG subclass항체의 분포)

  • Kim, Young-Jung;Hwang, Eung-Soo;Kang, Jae-Seung;Cha, Chang-Yong;Chang, Woo-Hyun;Kim, Yoon-Won;Cho, Min-Ki;Min, Chang-Hong
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.447-453
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    • 1986
  • To diagnose the typhoid fever rapidly and accurately in clinically suspected patients, the levels of IgG subclass antibody were measured by enzyme-linked immunosorbent assay(ELISA). With symptom, blood culture and agglutination test, tested persons were categorized into 6 groups as typhoid fever, FUO, paratyphi A or B, other bacterial infctions, cancers, and control. ELISA was performed on the polyvinyl chloride plates coated with killed whole cell($10^8\;cell/ml$) of S. typhi 0901W by poly-L-lysine applied as binding substance (and polyvinyl chloride as solid phase). The distribution of the level of IgG subclass antibodies in each group was analyzed and compared with other groups. The results obtained were summarized as follow: 1. The optimal dilution of the sera from patients with typhoid fever was 1:160, and those of the sheep anti-human IgG subclass and the peroxidase conjugated rabbit anti-sheep IgG were 1:4000 and 1:5000, respectively. 2. The absorbance levels of IgG subclass in the sera of typhoid fever patients were as follows; a) IgG1 value is $0.439{\pm}0.110$ b) IgG2 value is $0.416{\pm}0.165$ c) IgG3 value is $0.449{\pm}0.145$ d) IgG4 value is $0.525{\pm}0.154$ IgG subclass levels in the sera of typhoid patients were much higher than in control group and patient with paratyphi A or B as well as other infectious diseases. The sensitivity and the specificity in differential diagnosis of typhoid fever and other febrile diseases were 92% and 79% in the assay of IgG1 respectively, whereas those in the assay of IgG2 were 97% and 72%, respectively (above absorbance 0.3). 3. The absorbance levels of IgG subclass in the serial sera of typhiod fever patients tend to decrease to the level of absorbance 0.3 in 10 months from the onset of illness. 4. The order of absorbance levels of IgG subclass in the serum of each group were typhoid fever, paratyphi A or B, other infectious diseases, control and cancer. 5. For the serodiagnosis of typhoid fever against other febrile diseases, the sensitivity and the specificity in the assay of IgG2 activity were 76% and 93% in absorbance 0.4, respectively. 6. In the distribution of the level of each IgG subclass in the sera of FUO patients which were suspected of typhoid fever, the positive rate was ranged from 36% to 82%. This suggest that more than 50% of FUO patients are caused by S. typhi.

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Longitudinal Study of the Subgingival Microbial Change after Tetracycline Topical Application (Tetracycline 국소도포가 치은연하 세균분포에 미치는 영향)

  • Choi, Kwang-Choon;Lee, Young-Hee;Lee, Jin-Yong;Chung, Chong-Pyoung;Son, Seong-Heui
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.503-513
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    • 1986
  • Previous studies have developed the technique of topical application of tetracycline(TC) into the periodontal pockets and examined the change of clinical parameters and subgingival microbial morphotypes. The purpose of this study was to longitudinally examine the clinical and microbiological effects of topically applied TC in a double-blind and split-mouth design. Thirteen patients with moderate periodontitis, who were treated with or without TC application and scaling treatment, were examined. TC gel(3%) was used to apply into the selected periodontal pockets twice a week for 2 weeks. During the experiment, clinical parameters and subgingival microbial morphotypes were examined, and for isolation of black-pigmented Bacteroides(BPB) and streptococci, an anaerobic sample culturing was done at week 0, 2, and 7. In clinical observation the TC-scaled group exhibited a significant decrease of Gingival Inflammatory Index, Plaque Index, Sulcus Bleeding Index, pocket depth, and gingival crevicular fluid when compared to the TC-unsealed, placebo-scaled, and placebo-unsealed groups. The result of microbial morphotype observation showed a significant increase of coccal form and a decrease of spirochetes in the TC-scaled, TC-unscaled, and placebo-scaled groups. The culture study of streptococci revealed that TC with scaling treatment resulted in a significant increase of S. sanguis I at week 2, but its proportion had returned to the base line level. The anaerobic culture study showed that BPB was significantly reduced in the TC-scaled and TC-unsealed groups at week 7. Among BPB species, B. intermedius declined significantly with time treatment(week 2 and 7) in the TC-scaled and TC-unsealed groups. These results suggest that the settled pathogenic microflora can be succeeded by nonpathogenic microflora in periodontal pockets after TC treatment.

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Biological characteristics of Scuticociliate with a polymorphic life cycle of ciliate and cyst isolated from cultured olive flounder, Paralichthys olivaceus (양식넙치, Paralichthys olivaceus에서 분리된 Ciliate와 Cyst의 생활사를 갖는 스쿠티카섬모충의 생물학적 특성)

  • Jin, Chang-Nam;Kang, Hyun-Sil;Park, Geun-Tae;Lee, Je-Hee;Lee, Young-Don;Choi, Woo-Bong;Heo, Moon-Soo
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.19-28
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    • 2005
  • Scuticociliatosis has badly settled one of most damaging diseases during the seedling production process of olive flounder. Paralichthys olivaceus in Korea. We isolated a new type of Scuticociliate from flounder. The parasite metamorphoses to ciliate and cyst phases with each other by environmental changes and survive for a relatively long span. The ciliate was measured average 41.8 ${\mu}m$ in length and 21.0 ${\mu}m$ in width, and cyst was 17.0 ${\mu}m$ and 13.5 ${\mu}m$, respectively. Nutritional condition was determined as a major parameter of metamorphosing between ciliate and cyst stages. The ciliate transforms to a cyst stage because of food shortage, and the cyst returns to a ciliate stage with a favorite environmental condition and shows active growth and reproduction. The ciliate multiplied at the maximal density of $2.9 {\times} 10^5 {m\ell}^{-1}$cells in vitro cultivation at $15 ^{\circ}C$temperature using MS BHI medium and bacterial food sources. The ciliate could be proliferated at a 2.5 to $30 ^ \circ}C$ temperature range, pH 6 to 9, and 1 to 55 ppt salinity. Particularly, it survived over one week at $0 ^{\circ}C$temperature showing a high resistance against unfavorable environmental conditions. And the cyst survived for 320 days in the condition of $5 ^{\circ}C$with no feeding, but its survival period was markedly shortened in higher temperature conditions. The chemotherapeutants (formalin and hydrogen peroxide) were clarified as effective chemicals against the ciliate during in vitro trials, but the effect of therapeutants differed in proportion, depending upon the density and the bathing time of chemical compounds.

Carbon Dynamics of Plankton Communities in Paldang Reservoir (팔당호 플랑크톤 군집의 탄소생물량 동태)

  • Noh, Seong-You;Han, Myung-Soo
    • Korean Journal of Ecology and Environment
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    • v.41 no.2
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    • pp.174-187
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    • 2008
  • In an effort to identify structure and function of microbial loop in Paldang reservoir, we monitored environmental and biological factors at Kyungan stream (station K), Paldang dam (station P) and the confluence of North and South Han River (station M) from March to December, 2005. DOC concentration was higher in March to May and November than the others. Nutrient concentration in station K detected relatively higher than that of two stations. Both of phosphate and silicate gradually increased at all stations until September, after then decreased. The highest Chl-$\alpha$ concentration was observed at all stations in April, and November. The carbon biomass of bacteria and HNF were relatively higher in March, May and August than the others, whereas that of the ciliate showed no significant difference in monthly fluctuation. Nevertheless, the significant relationships revealed between ciliate (P<0.001) and HNF (P<0.05) and bacterial density. Tintinnopsis cratera, Didinium sp., Vorticella sp., Paramecium sp. and Strombidium sp. were dominant species in ciliate community. The dominant species of phytoplankton were Stephanodiscus hantzschii and Cyclotella meneghiniana at almost stations in Spring, Summer and Autumn. However, Aulacoseira granulata accounted for >95% of phytoplankton biomass at station P and M in Autumn. The carbon biomass of zooplankton was highest at station P and M in June, and relatively higher biomass observed at all stations in August, October and November. Diaphanosoma brachyurum and Bosmina longirostris were dominant in stations P and M of June and in all stations of October and November, respectively. The maximum growth (A. granulata: $0.17\;d^{-1}$, S. hantzschii: $0.14\;d^{-1}$) and grazing rate (A. granulata: 1.93 preys $d^{-1}$, S. hantzschii: 1.63 preys $d^{-1}$) of Bosmina longirostris revealed in algal preys as Aulacoseira granulata and Stephanodiscus hantzschii. In conclusion, these results suggest that bacteria and phytoplankton can play the most crucial source as prey within microbial food chain in Spring and Summer and grazing food chain in Autumn, respectively.

A Suvey on Satisfaction Measurement of Automatic Milking System in Domestic Dairy Farm (자동착유시스템 설치농가의 설치 후 만족도에 관한 실태조사)

  • Ki, Kwang-Seok;Kim, Jong-Hyeong;Jeong, Young-Hun;Kim, Yun-Ho;Park, Sung-Jai;Kim, Sang-Bum;Lee, Wang-Shik;Lee, Hyun-June;Cho, Won-Mo;Baek, Kwang-Soo;Kim, Hyeon-Shup;Kwon, Eung-Gi;Kim, Wan-Young;Jeo, Joon-Mo
    • Journal of Animal Environmental Science
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    • v.17 no.1
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    • pp.39-48
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    • 2011
  • The present survey was conducted to provide basic information on automatic milking system (AMS) in relation to purchase motive, milk yield and quality, customer satisfaction, difficulties of operation and customer suggestions, etc. Purchase motives of AMS were insufficient labor (44%), planning of dairy experience farm (25%), better performance of high yield cows (19%) and others (6%), respectively. Average cow performance after using AMS was 30.9l/d for milk yield, 3.9% for milk fat, 9,100/ml for bacterial counts. Sixty-eight percentage of respondents were very positive in response to AMS use for their successors but 18% were negative. The AMS operators were owner (44%), successor (44%), wife (6%) and company worker (6%), respectively. The most difficulty (31%) in using AMS was operating the system and complicated program manual. The rate of response to system error and breakdown was 25%. The reasons for culling cow after using AMS were mastitis (28%), reproduction failure (19%), incorrect teat placement (12%), metabolic disease (7%) and others (14%), respectively. Fifty-six percentages of the respondents made AMS maintenance contract and 44% did not. Average annual cost of the maintenance contract was 6,580,000 won. Average score for AMS satisfaction measurement (1 to 5 range) was 3.2 with decrease of labor cost 3.7, company A/S 3.6, increase of milk yield 3.2 and decrease of somatic cell count 2.8, respectively. Suggestions for the higher efficiency in using AMS were selecting cows with correct udder shape and teat placement, proper environment, capital and land, and attitude for continuous observation. Systematic consulting was highly required for AMS companies followed by low cost for AMS setup and systematization of A/S.

A Late-Maturing and Whole Crop Silage Rice Cultivar 'Mogwoo' (만생종 총체사료용 벼 신품종 '목우')

  • Lee, Sang-Bok;Yang, Chang-Ihn;Lee, Jeom-Ho;Kim, Myeong-Ki;Shin, Young-Seop;Lee, Kyu-Seong;Choi, Yong-Hwan;Jeong, O-Young;Jeon, Yong-Hee;Hong, Ha-Cheol;Kim, Yeon-Gyu;Jung, Kuk Hyun;Jeung, Ji-Ung;Kim, Junhwan;Shon, Ji-Young
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.33 no.2
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    • pp.81-86
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    • 2013
  • 'Mogwoo', a new high yield and whole crop silage rice (Oryza sativa L.) cultivar, was developed by the rice breeding team of the National Institute of Crop Science, RDA, Suwon, Korea, from 1999 to 2009, and was released in 2010. It was derived in 1999 from a cross between Dasanbyeo, having a high yield, and Suweon431/IR71190-45-2-1. A promising line, SR25848-C99-1-2-1, selected by the pedigree breeding method, was designated the name of 'Suweon 519' in 2007. This cultivar has about 155 days of growth period from seeding to heading, and is tolerance to lodging, with erect pubescent leaves as well as a long and thick culm. This cultivar has the same number of tillers per hill and higher spikelet numbers per panicle compared to Nokyang. 'Mogwoo' has longer leaves compared with other Tongil-type varieties. This new variety is resistant to grain shattering, leaf blast, bacterial leaf blight, and small brown planthopper. The biomass yield of 'Mogwoo' was 1,956 kg/10a in a regional test over three years. The result shows that 'Mogwoo' is adaptable to central and south-east plain areas of Korea.

Rheology and Sensory Properties of Mozzarella Cheese Made using Concentrated Milk from Ultrafiltration (한와여과 농축유로 제조한 모짜렐라 치즈의 물성 및 관능적 품질)

  • Song, Kwang-Young;Lee, Jong-Ik;Kim, Dong-Hyeon;Park, Jun-Ho;Chon, Jung-Whan;Hyeon, Ji-Yeon;Seo, Kun-Ho;Yoon, Yoh-Chang
    • Journal of Dairy Science and Biotechnology
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    • v.30 no.1
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    • pp.1-10
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    • 2012
  • The objective of this study was to evaluate the rheology and sensory properties of Mozzarella cheese manufactured by the traditional method and by an ultrafiltration technique and to attempt to develop a new type of Mozzarella cheese. We noted 3 observations. (1) The initial pH observed for raw milk and retentate of ultrafiltration (CF 2 and CF 3) did not vary much. However, there was a significant difference in the pH at stretching. The initial TAwas significantly different between the raw milk and retentate obtained by ultrafiltration (CF 2 and CF 3). However, no differences were observed in TA at stretching. (2) The predicted yield (Y50), adjusted to 50% total solids (TS), was affected by the concentration factor, fat content, and addition of starter and rennet. The Y50 increased with the concentration factor. 3) Hardness, elasticity, cohesiveness, and brittleness were affected by fat content, and addition of starter and rennet. The hardness, elasticity, and brittleness of Mozzarella cheese made from the retentate of ultrafiltration (CF 2 and CF 3) were higher than in Mozarella cheese made by the traditional method; however, the cohesiveness decreased during storage at $4^{\circ}C$ over a period of 3 months. (4) Flavor, body texture, appearance, color, and firmness decreased with bacterial count, fat content, and addition of starter and rennet. The results of the sensory evaluation of the Mozzarella cheese made from the retentate of ultrafiltration (CF 2 and CF 3) changed during storage at $4^{\circ}C$ over a 3-month period.

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Dry Matter Yield of Japanese Millet (Echinochloa crusgalli var. Frumentacea 〈Roxb.〉 Wight), Chemical Properties and Microbial Population of Soil as Affected by the Application of Fermented Sawdust Swine Manure in Cheju Volcanic Ash Soil Area (제주화산회토양에서 발효 톱밥 돈분 시용이 피의 건물수량, 토양의 이화학적 성질 및 토양 미생물에 미치는 영향)

  • Kim M. C.;Kim T. G.;Lee J. E.;Moon B. C.
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.25 no.3
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    • pp.159-168
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    • 2005
  • This experiment was carried out during the period from May to October 1998 to determine the effect of fermented sawdust swine manure application (SSM) on the herbage production of Japanese millet and soil properties in the Cheju brown volcanic ash soil. The randomized block design (T1 : basic chemical fertilizer, N 200 kg/ha + $P_2O_5\;300 kg/ha+K_2O$ 200kg/ha ; T2 : 1/2 basic chemical fertilizer, N $100+P_2O_5\;150+K_2O$ 100kg/ha; T3 : 1/2 basic SSM, N 100kg/ha, T4:basic SSM, N 200kg/ha; T5:2 times basic SSM, N 400 kg/ha ; T6:4 times basic SSM, N 800 kg/ha) was used. At the same application level of nitrogen 200kg/ha, the application of $100\%$ chemical fertilizer (T1) had significantly lower dry matter yield than that of $50\%$ chemical fertilizer and $50\%$ SSM (T2) or $100\%$ SSM (T4)(p<0.01). Dry matter yield increased with an increase of SSM application to N 400kg/ha level but decreased at N 800 kg/ha level. P, K and Ca contents of Japanese millet tended towards to decrease with an increase of SSM level. The application of chemical fertilizer lowered P and K content of Japanese millet in comparison with that of SSM (p<0.05). pH, available phosphorus. exchangeable potassium, Ca and Mg content of soil studied showed a significant increase with an increase of SSM application level (p<0.05). However, at the same application level of N 200kg/ha, there was statistically no significant difference between chemical fertilizer and SSM in those contents of soil after experiment. The bacterial number of soil among microbial population increased with an increase of SSM level, in June, 1998 (p<0.05), but there was no regular tendency, in October. The rate of bacteria to fungus in soil had a tendency to decrease with an increase of SSM level. In conclusion, it is recommended to use N 400kg/ha of SSM or N 100 kg/ha of chemical fertilizer+N 100 kg/ha of SSM for Japanese millet.

Combination of Hydrophobic Filtration and Enrichment Methods for Detecting Bacillus cereus in Fresh-Cut Cabbage

  • Lee, Sujung;Choi, Yukyung;Lee, Heeyoung;Kim, Sejeong;Lee, Jeeyeon;Ha, Jimyeong;Oh, Hyemin;Lee, Yewon;Kim, Yujin;Yoon, Yohan;Lee, Soomin
    • Journal of Food Hygiene and Safety
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    • v.33 no.5
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    • pp.325-329
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    • 2018
  • This study developed a rapid detection method for Bacillus cereus in fresh-cut cabbages. Fresh-cut cabbage samples were inoculated at 1-, 2- and 3-Log CFU/g, and pathogens were enriched in tryptic soy broth containing 0.15% polymyxin B at $30^{\circ}C$, $37^{\circ}C$, and $42^{\circ}C$ to determine the detection limit and appropriate enrichment temperature for multiplex PCR detection. Enriched bacterial cells in enrichment broth were collected in a hydrophobic filter prior to DNA extraction for multiplex PCR. Filters were resuspended in distilled water, and DNA was extracted from the suspension. DNA samples were further analyzed by multiplex PCR. Detection limit of multiplex PCR was 5-Log CFU/mL. B. cereus cell counts were higher (P < 0.05) at $42^{\circ}C$ than other temperatures. Detection rate of 1-, 2-, and 3-Log CFU/g inoculated samples were 60%, 80%, and 100% after enrichment respectively. However, when enriched samples were filtered with hydrophobic membrane filter, detection rates became 100%, regardless of inoculation level. Results indicate a combination of enrichment with hydrophobic filtration improves rapid detection efficiency of B. cereus in fresh-cut cabbage by multiplex PCR.