• Title/Summary/Keyword: Real samples

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Distribution of Lawsonia intracellularis in livestock transport car of slaughterhouse, Korea (도축장 출하차량에서 Lawsonia intracellularis 분포율 조사)

  • Lee, Su-Ji;Lee, Hee-Seon;Seo, Ji-Soo;Kim, Tae-Gyeom;Jeong, Jae-Kyo
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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    • pp.245-250
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    • 2018
  • Lawsonia intracellularis is the pathogenic agent of porcine proliferative enteritis (PPE). The bacterial pathogen infects the intestinal crypt cells which causes hyperplasia of the infected cells and leads to the process of intestinal pathogenesis. PPE includes some clinical maninfestations, including acute hemorrhagic diarrhea with sudden death in growing pigs and porcine intestinal adenomatosis, to a chronic diarrhea with reduced productivity of the infected pigs. The purpose of the present studies were carried out to determine L. intracellularis in livestock transport car of slaughterhouse. Distribution of L. intracellularis in livestock transport car were conducted using real-time polymerase chain reaction (real-time PCR) testing method, total 300 samples. Of 300 samples, 119 (39.7%) were detected as positive to L. intracellularis in livestock transport car. In seasonal analysis, 42 (28.0%) out of 150 samples in spring and summer season. 77 (51.3%) out of 150 sample in autumn and winter season. In regional analysis, 53 (88.3%) out of 60 cars and the detection ratio showed that regional variation in livestock transport car.

Comparison of the Standard Culture Method and Real-time PCR for the Detection of Vibrio parahaemolyticus in Seafoods and Vegetables (해산식품과 채소에서 Vibrio parahaemolyticus 검출을 위한 배지배양법과 real-time PCR의 비교검증)

  • Chon, Jung-Whan;Hyeon, Ji-Yeon;Hwang, In-Gyun;Kwak, Hyo-Sun;Han, Jeong-A;Chung, Yun-Hee;Song, Kwang-Young;Seo, Kun-Ho
    • Korean Journal of Food Science and Technology
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    • v.42 no.3
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    • pp.355-360
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    • 2010
  • Vibrio parahaemolyticus (V. parahaemolyticus), which is commonly found in raw seafood, causes gastroenteritis in humans. Rapid and effective methods have been developed as culture methods require up to 5-7 days. In this study, real-time PCR was compared with the standard culture method for detecting V. parahaemolyticus in seafood and radish sprout samples. Five hundred grams of the samples were artificially contaminated with V. parahaemolyticus then divided into 20 samples. The samples were incubated in alkaline peptone water and then streaked onto thiosulfate-citrate-bile saltssucrose agar. Biochemical tests for suspicious colonies were performed using the API 20NE strip. In parallel, real-time PCR was performed targeting the toxR gene using the enrichment broth. The real-time PCR was sensitive in discriminating V. parahaemolyticus from other foodborne pathogens. The detection limit of the real-time PCR was $10^3\;CFU/mL$ in phosphate-buffered saline. Although the real-time PCR detected more positive samples (76 out of 180, 42%) than the culture method (66 out of 180, 37%), there was no significant statistical difference (p>0.05) between the two methods. In conclusion, real-time PCR assays could be an alternative to the standard culture method for detecting V. parahaemolyticus in seafood and radish sprouts, which has many advantages in terms of detection time, labor, and sensitivity.

Detection of Plasmodium vivax by Nested PCR and Real-Time PCR

  • Genc, Ahmet;Eroglu, Fadime;Koltas, Ismail Soner
    • Parasites, Hosts and Diseases
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    • v.48 no.2
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    • pp.99-103
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    • 2010
  • Malaria is endemic in the Cukurova region while it is sporadic in other regions of Turkey. Therefore, the laboratory and clinical diagnosis of malaria is important for the treatment of malaria. In this study, 92 blood samples that were taken from the suspected malaria patients for routine diagnosis in a period of 10 years between 1999 and 2009 were analyzed. All of these blood samples were examined by microscopic examinations using Giemsa-stained thick blood films, nested PCR, and real-time PCR. The sensitivity-specificity and positive-negative predictive values for these diagnostic tests were then calculated. It was found that the positive predictive values of microscopic examination of thick blood films, nested PCR, and real-time PCR were 47.8%, 56.5%, and 60.9% for malaria, respectively. The real-time PCR was found to have a specificity of 75% and sensitivity of 100%, while specificity and sensitivity of nested PCR was found 81.2% and 97.7% according to the microscopic examination of thick blood films, respectively.

Quantitative Detection of Salmonella typhimurium Contamination in Milk, Using Real-Time PCR

  • JUNG SUNG JE;KIM HYUN-JOONG;KIM HAE-YEONG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.6
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    • pp.1353-1358
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    • 2005
  • A rapid and quantitative real-time PCR was developed to target the invasion A (invA) gene of Salmonella spp. We developed quantitative standard curves based on plasmids containing the invA gene. Based on these curves, we detected Salmonella spp. in artificially contaminated buffered peptone water (BPW) and milk samples. We were able to determine the invA gene copy number per ml of food samples, with the minimum detection limit of $4.1{\times}10^{3}$ copies/ml of BPW and $3.3{\times}10^{3}$ copies/ml of milk. When applied directly to detect and quantify Salmonella spp. in BPW and milk, the present real-time PCR assay was as sensitive as the plate count method; however, copy numbers were one to two logs higher than the colony-forming units obtained by the plate count methods. In the present work, the real-time PCR assay was shown to significantly reduce the total time necessary for the detection of Salmonella spp. in foods and to provide an important model for other foodborne pathogens.

Identification of Hanwoo and Holstein meat using MGB probe based real-time PCR associated with single nucleotide polymorphism (SNP) in Melanocortin 1 receptor (MC1R) gene (소 모색관련 MC1R 유전자의 SNP와 관련한 MGB probe에 기초한 real-time PCR을 이용한 한우육과 Holstein육의 판별)

  • Park, Sung-Do;Kim, Tae-Jung;Lee, Jae-Il
    • Korean Journal of Veterinary Research
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    • v.45 no.1
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    • pp.25-28
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    • 2005
  • The melanocortin 1 receptor (MC1R) plays an important role in regulation of melanin pigment synthesis within mammalian melanocytes. Mutations within the gene encoding MC1R have been shown to explain coat color variations within several mammalian species including cattle. To develope a rapid and accurate method for the identification of Hanwoo meat, we performed a single nucleotide polymorphism (SNP) analysis in Melanocortin 1 receptor (MC1R) gene using TaqMan$^{(R)}$ MGB probe-based real-time PCR. Two specific probes (one for Hanwoo and the other for Holstein and Black angus) were designed. At the 5' end of 2 TaqMan$^{(R)}$ MGB probes, 6-carboxyfluorescein (FAM) was labeled for Hanwoo, and VIC for Holstein and Black angus. As a result, Hanwoo samples showed FAM-positive signal only, whereas other samples showed VIC-positive. This result suggests that the TaqMan$^{(R)}$ MGB probe based real-time PCR technique would be very accurate, easy and reproducible method to discriminate between Hanwoo meat and Holstein/Black angus meat.

Default Prediction for Real Estate Companies with Imbalanced Dataset

  • Dong, Yuan-Xiang;Xiao, Zhi;Xiao, Xue
    • Journal of Information Processing Systems
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    • v.10 no.2
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    • pp.314-333
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    • 2014
  • When analyzing default predictions in real estate companies, the number of non-defaulted cases always greatly exceeds the defaulted ones, which creates the two-class imbalance problem. This lowers the ability of prediction models to distinguish the default sample. In order to avoid this sample selection bias and to improve the prediction model, this paper applies a minority sample generation approach to create new minority samples. The logistic regression, support vector machine (SVM) classification, and neural network (NN) classification use an imbalanced dataset. They were used as benchmarks with a single prediction model that used a balanced dataset corrected by the minority samples generation approach. Instead of using prediction-oriented tests and the overall accuracy, the true positive rate (TPR), the true negative rate (TNR), G-mean, and F-score are used to measure the performance of default prediction models for imbalanced dataset. In this paper, we describe an empirical experiment that used a sampling of 14 default and 315 non-default listed real estate companies in China and report that most results using single prediction models with a balanced dataset generated better results than an imbalanced dataset.

Development of a real-time PCR method for detection and quantification of the parasitic protozoan Perkinsus olseni

  • Gajamange, Dinesh;Yoon, Jong-Man;Park, Kyung-Il
    • The Korean Journal of Malacology
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    • v.27 no.4
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    • pp.387-393
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    • 2011
  • The objective of this study was to develop a real-time PCR method for the rapid detection and quantification of the protozoan pathogen Perkinsus olseni using a TaqMan probe. For the standard, genomic DNA was extracted from $10^5$ in vitro-cultured P. olseni trophozoites, and then 10-fold serial dilutions to the level of a single cell were prepared. To test the reliability of the technique, triplicates of genomic DNA were extracted from $5{\times}10^4$ cells and 10-fold serial dilutions to the level of 5 cells were prepared. The standards and samples were analyzed in duplicate using an $Exicycler^{TM}$ 96 real-time quantitative thermal block. For quantification, the threshold cycle ($C_T$) values of samples were compared with those obtained from standard dilutions. There was a strong linear relationship between the $C_T$ value and the log concentration of cells in the standard ($r^2$ = 0.996). Detection of DNA at a concentration as low as the equivalent of a single cell showed that the assay was sensitive enough to detect a single cell of P. olseni. The estimated number of P. olseni cells was similar to the original cell concentrations, indicating the reliability of P. olseni quantification by real-time PCR. Accordingly, the designed primers and probe may be used for the rapid detection and quantification of P. olseni from clam tissue, environmental water, and sediment samples.

Temporal Changes in Abundances of the Toxic Dinoflagellate Alexandrium minutum (Dinophyceae) in Chinhae Bay, Korea

  • Park, Tae-Gyu;Kang, Yang-Soon
    • Journal of Environmental Science International
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    • v.18 no.12
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    • pp.1331-1338
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    • 2009
  • Marine dinoflagellate Alexandrium minutum producing paralytic shellfish toxins is responsible for paralytic shellfish poisoning (PSP). To investigate its temporal distributions in Chinhae Bay where PSP occurs annually, SYBR Green I based A. minutum-specific real-time PCR probe was developed on the LSU rDNA region. Assay specificity and sensitivity were tested against related species, and its specificity was further confirmed by sequencing of field-derived samples. Ten months field survey in 2008 (a total 100 surface water samples) by using the real-time PCR probe showed that A. minutum was detected at very low densities of 1-4 cells $L^{-1}$ in May and June being spring in Chinhae Bay, Korea.

Disjoint Particle Filter to Track Multiple Objects in Real-time

  • Chai, YoungJoon;Hong, Hyunki;Kim, TaeYong
    • KSII Transactions on Internet and Information Systems (TIIS)
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    • v.8 no.5
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    • pp.1711-1725
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    • 2014
  • Multi-target tracking is the main purpose of many video surveillance applications. Recently, multi-target tracking based on the particle filter method has achieved robust results by using the data association process. However, this method requires many calculations and it is inadequate for real time applications, because the number of associations exponentially increases with the number of measurements and targets. In this paper, to reduce the computational cost of the data association process, we propose a novel multi-target tracking method that excludes particle samples in the overlapped predictive region between the target to track and marginal targets. Moreover, to resolve the occlusion problem, we define an occlusion mode with the normal dynamic mode. When the targets are occluded, the mode is switched to the occlusion mode and the samples are propagated by Gaussian noise without the sampling process of the particle filter. Experimental results demonstrate the robustness of the proposed multi-target tracking method even in occlusion.

Detection of Heterotrophic Dinoflagellate Pfiesteria piscicida (Dinophyceae) in Surface Water Samples Using Real-time PCR

  • Park, Tae-Gyu;Kang, Yang-Soon;Seo, Mi-Kyung;Park, Young-Tae
    • Fisheries and Aquatic Sciences
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    • v.11 no.4
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    • pp.209-211
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    • 2008
  • Heterotrophic dinoflagellate Pfiesteria piscicida (Dinophyceae) has been claimed to produce potent ichthyotoxins that cause disorientation and eventually death of fish and other marine animals. A real-time PCR probe targeting for SSU rRNA gene was used for detection of P. piscicida in Chinhae Bay, Korea. PCR inhibitors were successfully removed by dilution of template DNA. Positive detections were shown from surface water samples indicating the presence of P. piscicida in Chinhae Bay.