• 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.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.10
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• pp.1487-1492
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• 2014

This research applied and evaluated a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) using cytochrome b gene to detect pork contamination in meatballs from local markets in Surabaya and Yogyakarta regions, Indonesia. To confirm the effectiveness and specificity of this fragment, thirty nine DNA samples from different meatball shops were isolated and amplified, and then the PCR amplicon was digested by BseDI restriction enzyme to detect the presence of pork in meatballs. BseDI restriction enzyme was able to cleave porcine cytochrome b gene into two fragments (131 bp and 228 bp). Testing the meatballs from the local market showed that nine of twenty meatball shops in Yogyakarta region were detected to have pork contamination, but there was no pork contamination in meatball shops in Surabaya region. In conclusion, specific PCR amplification of cytochrome b gen and cleaved by BseDI restriction enzymes seems to be a powerful technique for the identification of pork presence in meatball because of its simplicity, specificity and sensitivity. Furthermore, pork contamination intended for commercial products of sausage, nugget, steak and meat burger can be checked. The procedure is also much cheaper than other methods based on PCR, immunodiffusion and other techniques that need expensive equipment.

• Journal of Preventive Medicine and Public Health
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• v.38 no.4
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• pp.420-424
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• 2005

Objectives : A few culture-confirmed cases of S. sonnei have been notified from Korean hospitals. The source of epidemic can't be firmly determined in such cases because of the rarity of this illness in the local communities and the timing of the outbreaks. The objective of this study is to estimate the source of epidemic by investigating the patients' lifestyles. Methods : Alibi verification was used to access the presumed source of the epidemic. PCR (Polymerase Chain Reaction) was used to rapidly detect the genes of Shigella in water specimens. Results : The common lifestyle trait among the Shigella infected patients was connected with Mt. Martyr in J city, Korea. The first patient's son had gone on a pilgrimage to Mt. Martyr with 41 friends and he had only eaten rice cakes on April 5th; the second patients had visited Mt. Martyr with their mother for a picnic on April 12th; the third patient had visited Mt. Martyr with 22 friends for a pilgrimage and the patient had only drunk holy water on April 13th. Therefore, the holy water of Mt. Martyr was reckoned to be the source of the epidemic. PCR detected the genes of Shigella two days before the S. sonnei was confirmed. Conclusion : The patients' lifestyles for 7 days before the onset of symptoms should be determined in terms of time, place and contacted people to find the source of infection when cases with food poisoning are seen in the hospital setting.

• Animal Bioscience
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• v.34 no.12
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• pp.1995-2002
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• 2021

Objective: Detection of adulteration in processed meats is an important issue for some countries due to substitution of beef with a cheaper source of protein like poultry. In this study, the presence of chicken meat was investigated using real-time polymerase chain reaction (real-time PCR) and enzyme-linked immunosorbent assay (ELISA) techniques to verify adulteration of fermented sausage samples. Methods: A total of 60 commercial samples were collected from 20 establishments in three replicates including 10 fermented sausage manufacturers and 10 butchers to investigate the presence of chicken meat with the sequential use of real-time PCR and ELISA techniques. In addition, pH, moisture content, water activity and color values of the samples were determined. Results: Both real-time PCR and ELISA showed agreement on the presence or absence of chicken meat in 55 out of 60 fermented sausage samples and chicken meat was identified with both methods in 16 samples. Five samples produced inconsistent results for the presence of chicken meat in the first run. Nevertheless, the presence of chicken meat was verified with both methods when these samples were analyzed for the second time. In addition, the average physico-chemical values of the fermented sausage samples tested positive for chicken meat were not significantly different from some of those fermented sausage samples tested negative for the chicken meat. Conclusion: The sequential use of real-time PCR and ELISA techniques in fermented sausages could be beneficial for the government testing programs to eliminate false negatives for detection of adulteration with chicken meat. Furthermore, consumers should not rely on some of the quality cues including color to predict the adulteration of fermented sausages with chicken meat since there were no statistical differences among some of the samples tested positive and negative for chicken meat.

• The Korean Journal of Food And Nutrition
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• v.13 no.1
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• pp.1-5
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• 2000

Detection for heat-labile enterotoxin(LT) of enterotoxigenic Escherichia coli(ETEC, EC81, O148:H28) and enterotoxin of enterotoxigentic Clostridium perfringents type A(CP, NCTC8238, Hobbs serotype 2) by use of a polymerase chain reaction (PCR) assay were positive reaction, which using LT gene-specific primers of ETEC with a detection limit equivalent from 100ng/${\mu}\ell$ to 1 pg of a DNA fragment of 417-bp in EC81 and enterotoxin gene-specific primers of CP with a detection limit equivalent from 100ng/${\mu}\ell$ to 10pg of a DNA fragment of 364-bp in NCTC8238. Detection for a LT gene of ETEC highly appeared 10-fold sensitivity than an enterotoxin gene of CP.

• Clinical and Experimental Reproductive Medicine
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• v.21 no.3
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• pp.281-284
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• 1994

Sex determination in genomic DNA from human blood leucocytes was performed by amplification of human Y chromosome-specific DNA sequences using PCR technique. A clear DNA fragment(154 nucleotides long) was appeared only in the male genomic DNA, but no specific band was observed in the case of female genomic DNA and negative control. To know the sensitivity of this method, the amplification reaction was performed in genomic DNA diluted to 2pg equivalent to the amonut present in the single human cell, and clear band also observed. The PCR amplification was so succesfully performed in the single leucocyte separated from human blood using micromanipulator that this techniqe is assumed to be applied to single blstomere before embryo transfer.

• Korean Journal of Environmental Biology
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• v.35 no.4
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• pp.468-475
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• 2017

This study is for the consideration of the existence tendency of Kudoa septempunctata in olive flounder. In general, muscle has shown a strong PCR positive reaction in spores containing tissues rather than non-containing tissues. However, blood PCR results showed opposed tendency. In various organs of the tested fish containing spores in muscle tissue, heart had shown positive reaction along with muscle at PCR analysis. Muscle fiber necrosis was observed at the histological observation, and this degeneration was common in both samples. The one sample was the PCR positive muscle containing spore and the other was the PCR positive muscle non-containing spore. Both of muscle tissues indicated a positive reaction at ISH (in-situ hybridization) against K. septempunctata.

• BMB Reports
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• v.35 no.2
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• pp.248-250
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• 2002

Discrimination between the amplification of mRNA and contaminating genomic DNA is a common problem when performing a reverse transcriptase-polymerase chain reaction (RT-PCR). Even after treatment of the samples with DNAse, it is possible that negative controls (samples in which no reverse transcriptase was added) will give positive results. This indicates that there was amplification of DNA, which was not generated during the reverse transcriptase step. The possibility exists that Taq DNA polymerase acts as a reverse transcriptase, generating cDNA from RNA during the PCR step. In order to test this hypothesis, we incubated samples with a DNAse-free RNAse after the cDNA synthesis. Comparison of the results that were obtained from these samples (incubated with or without DNAse-free RNAse) confirms that the reverse transcriptase activity of Taq DNA polymerase I is a possible source of false positive results when performing RT-PCR from intronless genes. Moreover, we describe here a simple and rapid method to overcome the false positive results that originate by this activity of Taq polymerase.

• Biomedical Science Letters
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• v.4 no.2
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• pp.129-135
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• 1998

We have carried out polymerase chain reaction (PCR) to investigate if retropseudogene for CYP2El is present in human genome. PCR primers were designed based on the structure of functional CYP2El gene and used to amplify both functional gene and retropseudogene in one reaction. From the repeated experiments we were able to amplify a previously unidentified CYP2El retropseudogene that was present in human genome. Its detailed structure was confirmed by Southern blotting and DNA sequencing. Nucleotide sequence of this retropseudogene was completely matched up to human liver CYP2El mRNA suggesting that the development of this retropseudogene might be a relatively recent event.

• Journal of Korean Society of Forest Science
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• v.81 no.4
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• pp.320-324
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• 1992

It has been reported that there are two distinct phenotypes in Fraxinus mandshurica Rupr. growing in Korea. Recently developed polymerase chain reaction(PCR) was used to detect DNA sequence polymorphism in the species. Using a thermostable DNA polymerase and synthetic DNA primers, unknown DNA sequences from the species were randomly amplified. The two types of the species produced different DNA amplification pattern with three different primers tested. Although DNA polymorphism was detected among individuals within types, each type has its own distinct pattern. The two types could be easily differentiated by trier characteristic predominant bands.