• Title/Summary/Keyword: Screening of Salmonella spp

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Rapid Screening of Salmonella spp. Using PBM BioSignTM Salmonella Test and Evaluation of the PBMS Test

  • Lim, J.Y.;Kwon, N.H.;Kim, J.M.;Jung, W.K.;Park, K.T.;Hong, S.K.;Park, Y.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.17 no.12
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    • pp.1746-1750
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    • 2004
  • The PBM ${BioSign}^{TM}$ Salmonella (PBMS) test kit based on an mmunochromatographic method was evaluated for the screening of Salmonella spp. in pure cultures, and 80, 15, and 10 artificially and naturally contaminated, and negative controlled food samples, respectively. The PBMS test involves presumptive qualitative procedures, detecting the presence of Salmonella spp. in foods within 26 h total testing period and allowing the user to release negative products 70 h earlier than the conventional methods. The PBMS test using Buffered Peptone Water and Rappaport-Vassiliadis broth was evaluated for 10 different food types for various Salmonella spp. It showed detection limits of 1 to 25 colony forming units (CFU)/25 g. No cross-reaction was observed, particularly to other gramnegative bacteria. These results indicate the PBMS test is a rapid and inexpensive procedure for the screening of Salmonella spp. present at low concentrations (1 to 25 CFU/25 g) in foods.

Rapid and Sensitive Detection of Salmonella spp. by Using a Loop-Mediated Isothermal Amplification Assay in Duck Carcass Sample (오리 도체에서 등온유전자증폭기법을 이용한 Salmonella spp. 신속 고감도 검출 기법 연구)

  • Cho, Ae-Ri;Dong, Hee-Jin;Cho, Seongbeom
    • Food Science of Animal Resources
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    • v.33 no.5
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    • pp.655-663
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    • 2013
  • In this study, a rapid and sensitive detection tool for screening Salmonella spp. by using a loop-mediated isothermal amplification (LAMP) assay targeting the genomic sequence of the invA gene was developed. The inclusivity and exclusivity were both at 100% in the analysis, and the limit of detection (LOD) in a pure S. Enteritidis culture suspended in saline was $3.2{\times}10^3$ CFU/mL at 18.17 min ($R^2$ = 0.9446). The LODs of the LAMP assay in buffered peptone water with Salmonella (BPW) and duck carcass swab sample enriched in BPW with Salmonella (BPWS) after 0 and 12 h incubation were $3.2{\times}10^3$ CFU/mL and $3.2{\times}10^0$ CFU/mL, respectively. Comparing the LODs in BPW with those in BPWS, the LAMP assay was less influenced by compounds of duck carcass swab sample than the PCR assay. Sensitivity and specificity of the LAMP assay in 50 duck carcass swab samples enriched in BPW for 6 h were 96% and 84%, respectively, indicating that the LAMP assay is a rapid, simple and sensitive assay, which can be utilized as a potential screening tool of Salmonella spp. in duck carcass sample.

Development of a lateral flow dipstick test for the detection of 4 strains of Salmonella spp. in animal products and animal production environmental samples based on loop-mediated isothermal amplification

  • Wirawan Nuchchanart;Prapasiri Pikoolkhao;Chalermkiat Saengthongpinit
    • Animal Bioscience
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    • v.36 no.4
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    • pp.654-670
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    • 2023
  • Objective: This study aimed to develop loop-mediated isothermal amplification (LAMP) combined with lateral flow dipstick (LFD) and compare it with LAMP-AGE, polymerase chain reaction (PCR), and standard Salmonella culture as reference methods for detecting Salmonella contamination in animal products and animal production environmental samples. Methods: The SalInvA01 primer, derived from the InvA gene and designed as a new probe for LFD detection, was used in developing this study. Adjusting for optimal conditions by temperature, time, and reagent concentration includes evaluating the specificity and limit of detection. The sampling of 120 animal product samples and 350 animal production environmental samples was determined by LAMP-LFD, comparing LAMP-AGE, PCR, and the culture method. Results: Salmonella was amplified using optimal conditions for the LAMP reaction and a DNA probe for LFD at 63℃ for 60 minutes. The specificity test revealed no cross-reactivity with other microorganisms. The limit of detection of LAMP-LFD in pure culture was 3×102 CFU/mL (6 CFU/reaction) and 9.01 pg/μL in genomic DNA. The limit of detection of the LAMP-LFD using artificially inoculated in minced chicken samples with 5 hours of pre-enrichment was 3.4×104 CFU/mL (680 CFU/reaction). For 120 animal product samples, Salmonella was detected by the culture method, LAMP-LFD, LAMP-AGE, and PCR in 10/120 (8.3%). In three hundred fifty animal production environmental samples, Salmonella was detected in 91/350 (26%) by the culture method, equivalent to the detection rates of LAMP-LFD and LAMP-AGE, while PCR achieved 86/350 (24.6%). When comparing sensitivity, specificity, positive predictive value, and accuracy, LAMP-LFD showed the best results at 100%, 95.7%, 86.3%, and 96.6%, respectively. For Kappa index of LAMP-LFD, indicated nearly perfect agreement with culture method. Conclusion: The LAMP-LFD Salmonella detection, which used InvA gene, was highly specific, sensitive, and convenient for identifying Salmonella. Furthermore, this method could be used for Salmonella monitoring and primary screening in animal products and animal production environmental samples.

Modified sorbitol MacConkey agar for the rapid isolation of Escherichia coli O157:H7

  • Jung, Byeong-yeal;Jung, Suk-chan;Lee, Na-kyung;Cho, Seong-kun;Cho, Dong-hee;Her, Moon;Yoon, Yong-dhuk;Kim, Bong-hwan
    • Korean Journal of Veterinary Research
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    • v.39 no.4
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    • pp.765-771
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    • 1999
  • Unlike most Escherichia coli strains, E coli O157 : H7 didn't ferment sorbitol within 24h of incubation and showed a negative reaction for $\beta$-glucuronidase. We developed a new medium for the rapid isolation of E coli O157 : H7 using sorbitol MacConkey agar with cefixime, potassium tellurite and 4-methylumbelliferyl-${\beta}$-D-glucuronide (MUG) as a primary plating medium. The addition of $20{\mu}g/ml$ of vancomycin in enrichment broth for E coli O157 : H7 inhibited lots of Gram positive bacteria. Three strains (10.3%) of 29 non-O157 E coli strains and 3 strains (8.3%) of 36 Salmonella spp were inhibited at the $0.05{\mu}g/ml$ of cefixime and 23 strains (79.3%) of 29 non-O157 E coli strains and 12 strains (33.3%) of 36 Salmonella spp were inhibited at the $2.0{\mu}g/ml$ of potassium tellurite. But none of the E coli O157 : H7 was affected at these concentration. The addition of MUG at $100{\mu}g/ml$ level to sorbitol MacConkey agar with cefixime and potassium tellurite (CTM-SMAC) aided in the rapid isolation of E coli O157 : H7 from samples by checking sorbitol-negative and $\beta$-glucuronidase negative phenotypes simultaneously. In conclusion, inoculation of a positive in the O157 screening test from enrichment broth on CTM-SMAC appeared to be a rapid, cost-effective and sensitive method for the isolation of E coli O157 : H7.

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Screening of Bifidobacterium spp. for the Development of Infant Probiotics (유아용 생균제 개발을 위한 Bifidobacterium spp.의 선발)

  • Yang, Hyun-Ju;Jang, Keum-Il;Kim, Chung-Ho;Lee, Yoon-Bok;Sohn, Heon-Soo;Kim, Kwang-Yup
    • Korean Journal of Food Science and Technology
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    • v.36 no.5
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    • pp.790-794
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    • 2004
  • Bifidobacterium spp. exhibits the highest number of counts among species of microflora in breast-feeding infant intestines and has been used as probiotics. From infant groups with different diets, 42 Bifidobacterial strains were isolated by selective plate, Gram-staining, and morphology using method of Mitsuoka, among which seven isolates were identified as Bifidobacterium spp. by F6PPK test, MIDI, and PCR. B. bifidum PBH-30, selected for development of probiotics, showed high resistance against low pH and oxgall treatment, and inhibition against pathogens such as Salmonella typhimurium and Staphylococcus aureus. B. bifidum PBH-30 could be applicable to dairy products as probiotic strains due to its excellent growth in raw milk.

Development and Evaluation of an Immunochromatographic Assay for Screening Listeria spp. in Pork and Milk

  • Kim, Seong-Hee;Kim, Jin-Young;Han, Woong;Jung, Byeong-Yeal;Chuong, Pham-Due;Joo, Hae-Jin;Ba, Hoa-Van;Son, Won-Geun;Jee, Young-Heun;Yoon, Byoung-Su;Lee, Yong-Soon;Lim, Yoon-Kyu
    • Food Science and Biotechnology
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    • v.16 no.4
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    • pp.515-519
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    • 2007
  • Rapid immunochromatographic assay (ICA) kits were developed using flagella-specific monoclonal antibodies (MAbs) and rabbit polyclonal antibodies for screening Listeria spp. in food. The establishment of different formats, MAb 2B1 as capture antibody and MAb 7A3 or rabbit polyclonal antibodies as detector antibody, was compared. The 2 formats of the ICA kit were shown to have specific reactions with Listeria and no cross-reactivity with any of the non-Listeria including Escherichia coli O157:H7 and Salmonella enteritidis. The detection limits of the ICA kit using the combination of gold-labeled MAb 7A3 and MAb 2B1 showed $1{\times}10^5$ and $1{\times}10^6\;CFU/0.1\;mL$ at 22 and $30^{\circ}C$, respectively. The other format of the ICA kit using the combination of gold-labeled rabbit polyclonal antibodies and MAb 2B1 showed $1{\times}10^6\;CFU/0.1\;mL$ at $22^{\circ}C$ but weak signal at 30 culture. The format utilizing MAb was more sensitive than the one using polyclonal antibodies for capture antibody. Samples contaminated with L. monocytogenes 4b culture (9-10, 5-6, and 1-2 CFU/mL) on pork and pasteurized milk were confirmed as positive results. Current data suggests that this ICA kit is a rapid, simple and effective tool to screen for Listeria spp. in food.

Microbiological Hazard Analysis of Botanical Raw Materials used for Functional Health Foods and Preliminary Screening for Irradiation of Aloe Powder Products (건강기능식품 중 주요 식물성 원료에 대한 미생물학적 위해 분석과 알로에 제품에 대한 방사선 조사 가능성 검지)

  • Sung, Dong-Eun;Lee, Jee-Hye;Oh, Sang-Suk
    • Journal of Food Hygiene and Safety
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    • v.22 no.1
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    • pp.15-22
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    • 2007
  • Health functional foods refer to food which manufactured in the form of a tablet, capsule, granule or pill, using materials and ingredients with useful function for a human body in Korea. It needs to be confirmed as safe. Microbiological analyses on 37 samples of botanical raw materials used in the health functional food were performed. Microbiological analyses and probability of irradiation using PSL on 4 samples of aloe powder products were studied. In health functional food ingredients APC was $10^{3}-10^{6}\;CFU/g$ and coliform counts were $10^{2}-10^{5}\;CFU/g$. Among 37 samples B. cereus were found in 12 samples. Four samples of aloe powder products were tested for possible irradiation using preliminary PSL, Two samples of aloe powder products showed positive on preliminary PSL test for irradiation.

Analysis on Hazard Microorganisms in Raw Materials and Processing Environment for Sunsik Manufacture (선식용 곡류원료 및 제조공정에 따른 유해미생물 오염도 분석)

  • Kim, Jin-Hee;Lee, Yu-Keun;Yang, Ji-Young
    • Journal of Food Hygiene and Safety
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    • v.26 no.4
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    • pp.410-416
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    • 2011
  • Cereals are the main raw material for sunsik manufacture. As the fundamental processing step, it is very important to confirm the level of the microorganism contamination in the cereals. This study was carried out a micrbiological screening of cereal samples for sunsik from 19 companies located in South Korea. Ten kinds of cereals which were glutinous rice, barley, brownrice, blackbean, blackrice, blacksesame, sorghum, millet, perilla seed, and adlay were investigated. As the results, the contaminations of general bacterial were 3.1~8.6 log(CFU/g). The results of Escherichia coli were 1.0~3.4 log(CFU/g). There was no contamination of Salmonella. spp in any cereal samples except black sesame and mold was detected in barley. The experiment for microbiological contamination during sunsik processing was also investigated in this study. The results of general bacteria were detected as 5.1~8.5, 4.4~7.5, 1.0~2.3, 2.4~4.2, 1.0~4.0, 3.4~4.2, 4.3~5.2, and 3.3~5.5 log(CFU/g) during environment of warehousing, washing, steaming, 1st cooling, drying, 2nd cooling, grinding, and packaging process, respectively. The results of coliform were 1.0~2.0 log(CFU/g) during warehousing respectively. Mold was found in warehousing. In case of the instruments, the contaminations of general bacterial were 4.2~7.5, 0.1~2.0, 0.1~3.2, 3.7~4.0, 2.5~3.0, and 3.8~5.2 log(CFU/g) in cereals tanks, washing machines, grinding machines, packaging machines, and workrooms. The results of coliform were 2.4~4.0, 0.0~4.1 log(CFU/g) in cereals tanks and grinding machines, respectively. Mold were only found in cereals tanks, grinding machines, and workings. Therefore, the risk of hazard microorganisms contmination might be decrased as the exhaustive management is applied to the whole sunsik process.