• Preventive Nutrition and Food Science
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• v.7 no.4
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• pp.373-377
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• 2002

Allergens in processed foods may place persons with food allergies at significant risk when the labels do not Provide sufficient warnings or identification of high-risk ingredients. Because egg proteins are common food allergens, this study was carried out to identify hen's egg albumin (ovalbumin, OVA) in five commercially processed foods containing egg (custayd, cookie and pasta), and chicken meat (sausage and meatball) by immunological methods using commercially produced murine monoclonal immunoglobulin G (M-IgG), immunoblotting and enzyme linked immunosorbent assay (ELISA). Sample buffer with chelating and reducing agents was prepared and used for the preparation of the protein fractions from the foods. Most bands in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) profile (5~15% gradient gel) presented at 75 kDa below. OVA (43 kDa) in the sample lanes could not be visually observed on the gel. However, OVA in solutions prepared from custard and cookie could be detected by M-IgG, but were not detected in sausage and pasta. OVA in all samples could be quantitatively determined by the equation obtained from the standard curve by ELISA. Cookie and custard containing egg white and egg, respectively, contained very high concentrations of OVA. OVA in the other products were present in relatively low concentrations, but sufficiently high to pose possible risk of allergy, ELISA is a very sensitive and precise method for the identification and quantification of allergens in food products including allergy-inducible materials.

• Journal of Microbiology and Biotechnology
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• v.19 no.1
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• pp.108-112
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• 2009

A simplified method for the purification of cholera toxin was developed. The 569B strain of Vibrio cholerae, a recognized hyper-producer of cholera toxin, was propagated in a bioreactor under conditions that promote the production of the toxin. The toxin was separated from the bacterial cells using 0.2-${\mu}m$ crossflow microfiltration, the clarified toxin was passed through the membrane into the permeate, and the bacterial cells were retained in the retentate. The 0.2-${\mu}m$ permeate was then concentrated 3-fold and diafiltered against 10 mM phosphate buffer, pH 7.6, using 30-kDa crossflow ultrafiltration. The concentrated toxin was loaded onto a cation exchange column, the toxin was bound to the column, and most of the impurities were passed unimpeded through the column. The toxin was eluted with a salt gradient of phosphate buffer, pH 7.0, containing 1.0 M NaCl. The peak containing the toxin was assayed for cholera toxin and protein and the purity was determined to be 92%. The toxin peak had a low endotoxin level of $3.1\;EU/{\mu}g$ of toxin. The purified toxin was used to prepare antiserum against whole toxin, which was used in a $G_{M1}$ ganglioside-binding ELISA to determine residual levels of toxin in an oral inactivated whole-cell cholera vaccine. The $G_{M1}$ ganglioside-binding ELISA was shown to be very sensitive and capable of detecting as little as 1 ng/ml of cholera toxin.

• Journal of Veterinary Clinics
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• v.30 no.4
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• pp.230-235
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• 2013

Johne's disease is a chronic inflammatory bowel disease of cattle, sheep, goats and other ruminants. Mycobacterium paratuberculosis is the etiologic agent of this disease. Many studies have been carried out on paratuberculosis from Korean native cattle and dairy cattle in multiple areas around nation, but there is no report in Busan area. The purpose of this study is to investigate the seroprevalence of bovine paratuberculosis in Busan area from March in 2011 to October in 2012. A total of 863 Korean native cattle of 213 farms were tested by ELISA method. The 287 (33.3%) Korean native cattle of 119 (55.9%) farms were positive in ELISA. In regional analysis, 234 (33.6%) out of 696 cows in Kijang-gun, 35 (39.3%) out of 89 cows in Gangseo-gu and 15 (20.8%) out of 72 cows in Geumjeong-gu were positive. In sexual analysis, 277 (33.6%) out of 824 cows in Female and 10 (25.6%) out of 39 cows in Male were positive. In aga-related analysis, 13 (22.4%) out of 58 cows in 1 year, 33 (32.0%) out of 103 cows in 2 years, 87 (34.1%) out of 255 cows in 3 years, 118 (36.6%) out of 322 cows in 4 years, 21 (36.8%) out of 57 cows in 5 years, 8 (29.6%) out of 27 cows in 6 years, 6 (31.6%) out of 19 cows in 7 years and 1 (4.5%) out of 22 cows in 8-11 years were positive.

• Korean Journal of Veterinary Service
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• v.38 no.1
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• pp.43-49
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• 2015

Bovine viral diarrhea virus (BVDV) is a very important viral disease virus in cattle, domestic and wild ruminants. The purpose of this study is to investigate the positive rate of bovine viral diarrhea virus antigen by ELISA from Korean native and beef cattle reared in Busan area from March in 2013 to October in 2014. A total of 1,129 bovine blood samples were collected from 140 farms, 1,111 Korean native cattle of 135 farms and 18 beef cattle of 5 farms. Test for antigen was carried out by ELISA method. In general analysis, the positive rate of bovine viral diarrhea virus antigen were 0.7% (8/1,129) cattle and 5.0% (7/140) farm. In regional analysis, the positive rate of BVDV antigen of farm in Kijang-gun, Gangseo-gu, Geumjeong-gu, Saha-gu and Dongnae-gu were 1.4% (2/94), 3.6% (5/37), 0% (0/7), 0% (0/1) and 0% (0/1), respectively, and the positive rate of BVDV antigen of cattle were 0.4% (3/770), 1.5% (5/333), 0% (0/24), 0% (0/1) and 0% (0/1), respectively. The positive rate of BVDV antigen according to sex were 0.6% (6/1,085) female cattle and 4.6% (2/44) male cattle. According to the age of cattle, the positive rate of BVDV antigen in 1 year, 2 years, 3 years and 5 years old were 1.9% (4/215), 0.4% (1/265), 0.9% (2/234) and 1.0% (1/103), respectively, but 4 years (0/198), 6 years (0/55), 7 years (0/24), 8 years (0/14), 9 years (0/10), 10 years (0/7) and 11-15 years (0/3) old were negative, respectively.

• Food Science of Animal Resources
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• v.32 no.6
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• pp.706-712
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• 2012

The present study was aimed to produce a chicken polyclonal antibody against Cronobacter muytjensii and to develop an immunoassay for its detection. Purification of anti-C. muytjensii IgY from egg yolk was accomplished using various methods such as water dilution and salt precipitation. As a result, sodium dodecyl sulfate-polyacrylamide gel electrophoresis produced two bands around 30 and 66 kDa, corresponding to a light and a heavy chain, respectively. Indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was performed to determine the effectiveness of the chicken IgY against C. muytjensii. The optimum conditions for detecting C. muytjensii by indirect ELISA and checkerboard titration of the antigen revealed an optimum average absorbance at the concentration of 18 ${\mu}g/mL$, having ca. $10^8$ coated cells per well. The anti-C. muytjensii IgY antibody had high specificity for C. muytjensii and low cross-reactivity with other tested pathogens. In this assay, no cross-reactivity was observed with the other genera of pathogenic bacteria including Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, Bacillus cereus, Enterobacter aerogenes, Salmonella Enteritidis and Listeria monocytogenes. In addition, detection of C. muytjensii in infant formula powder showed a low matrix effect on the detection curve of IC-ELISA for C. muytjensii, with similar detection limit of $10^5$ CFU/mL as shown in standard curve. These findings demonstrate that the developed method is able to detect C. muytjensii in infant formula powder. Due to the stable antibody supply without sacrificing animals, this IgY can have wide applications for the rapid and accurate detection of C. muytjensii in dairy foods samples.

• Parasites, Hosts and Diseases
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• v.40 no.2
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• pp.75-81
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• 2002

We investigated population densities of mosquitoes infected with sporozoites in three highly epidemic areas of Josan-ri and Jangpa-ri (Paju City) and Dongjung-ri (Yeoncheon County) in Korea. Anopheline mosquitoes were collected front both indoors and outdoors by human baiting collection method during the period of the first week of June to the second week of September 1999. Total 13,296 female mosquitoes were collected and 8,650 (65.1%) were Anophelines. Thirty seven percent (3,199) of the Anopheline mosquitoes were captured outdoors and 63.9% (5,531) indoors. Employing a sandwich enzyme-linked immunosorbent assay (ELISA), we analyzed a total of 7,820 Anopheline mosquitoes and found that 7 Anopheline mosquitoes were infected with sporozoites. The positive rate in Josan-ri was 0.14% (5/3,500) and 0.15% (2/1,370) in Jangpa-ri. The total positive rate in all three surveyed areas was 0.09% (7/7,820). The mosquitoes infected with the sporozoites were detected on June $28^{th}$ (n=2), July $5^{th}$ (n=1), July $19^{th}$ (n=1), August $9^{th}$ (n=1), September $6^{th}$ (n=1), and the last one on September $13^{th}$ (n=1). They were all classified as Anopheles sinensis, which showed positive reaction in ELISA test. Therefore it might be concluded that Anopheles sinensis plays an important role in re-emerging malaria transmission in Korea.

• Korean Journal of Microbiology
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• v.43 no.3
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• pp.179-185
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• 2007

In this study, we used the method of guanidin isothiocyanate and boiling with Chelex-100 resin to extract genomic DNA of Ralstonia solanacearum from soil. It is more efficient than general protocols to remove inhibitory compounds in soil and R. solanacearum on. Then, we applied polymerase chain reaction and DNA enzyme-linked immunosorbent assay (ELISA) to identify and detect pathogen. The fliC gene of R. solanacearum was selected for specific detection of pathogen and primer sets were designed. Among the primer sets, two specific and sensitive primer sets, RsolfliC(forward: 5-GAACGCCAACGGTGCGAACT-3 and reverse; 5-GGCGGCCTTCAGGGAGGTC-3, designed by J. $Sch\ddot{o}nfeld$ et al.) and RS_247 (forward: 5-GGCGGTCTGTCGGCRG-3 and reverse; 5-CGGTCGCGTTGGCAAC-3 designed by this study), were designed to perform nested PCR. Nested PCR primer was labeled with biotin for hybridization between nested PCR product and probe to analyze with DNA ELISA.

• Korean Journal of Veterinary Service
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• v.37 no.1
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• pp.45-49
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• 2014

Pullorum disease and fowl typhoid are septicemic avian diseases transmitting through egg by transovarian infections. A series of tests has been performing in breeding flocks of chickens and test plans for proper inspection have been modified by government veterinary institute to control of such diseases. To improve inspection plans, different test methods were compared using fowl typhoid positive samples from a poultry farm located in Jeonbuk state in 2012. Based on first inspection, 11 samples among total 200 samples were positive by rapid slide agglutination (RSA) test and 7 samples among RSA positive samples were finally diagnosed as Salmonella Gallinarum infection by ELISA, bacterial isolation, PCR, and histopathologic examination. In the second inspection, 20 samples among total 100 samples were positive by RSA test. Among RSA positive ones, 19 samples were positive by ELISA, S. Gallinarm were successfully isolated in 3 samples, and 16 samples were positive by PCR in the cecal tonsils where were not successful for bacterial isolation. Based on histopathologic examination, severe inflammation in the 13 cecal tonsils and infiltration of lymphocytes and heterophils in the 11 livers were observed. Therefore, we suggest that bacterial isolation, PCR, and histopathologic examination methods in the third inspection need to be further used in various tissues for correct diagnosis and for final eradication of pullorum disease and fowl typhoid in breeding flocks of chickens.

• Korean Journal of Veterinary Research
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• v.45 no.1
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• pp.55-61
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• 2005

Serum samples of 1,175 pigs from 148 Korean swine farms not using Mycoplasma hyopneumoniae (M. hyo) vaccines were collected for seroepidemiological study of M. hyo infection by indirect ELISA method. Informations of each farm were provided about province where the farm was located and season when blood samples were collected. Then, the selected farms were divided into farm units which had 5 serum samples according to production stages : sow, suckling piglet (<30 days old), nursery pig (30-70 days old), and growing pig (>70 days old). Seroprevalence of M. hyo infection according to production stages, province, and season was investigated by using ELISA-positve rate of the selected samples for each study. This study showed that 85.34% (78.94-91.78%, 95% CI) of farms were positive to M. hyo infection and 34.81% (32.09-37.53%, 95% CI) among pigs were sero-positive to M. hyo infection in Korean swine farms. In the study of seroprevalence by production stage, most farms had sows and growing pigs which were sero-positive to M. hyo infection (sow: 83.05%, growing pigs: 87.72%) and most pigs seemed to be naturally infected by M. hyo at 8-10 weeks of age. Also, M. hyo infection showed seasonal pattern that most pigs were infected in late fall to early winter. However, in the study of seroprevalence by province, there was no significant correlation between province and M. hyo sero-positive rate.

• Journal of Microbiology and Biotechnology
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• v.21 no.7
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• pp.719-727
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• 2011

Botulism is a neuroparalytic disease caused by Clostridium botulinum, which produces seven (A-G) antigenically diverse neurotoxins (BoNTs). BoNTs are the most poisonous substances known to humans, with a median lethal dose ($LD_{50}$) of approximately 1 ng/kg of body weight. Owing to their extreme potency and lethality, they have the potential to be used as a bioterrorism agent. The mouse bioassay is the gold standard for the detection of botulinum neurotoxins; however, it requires at least 3-4 days for completion. Attempts have been made to develop an ELISA-based detection system, which is potentially an easier and more rapid method of botulinum neurotoxin detection. The present study was designed using a synthetic gene approach. The synthetic gene encoding the catalytic domain of BoNT serotype B from amino acids 1-450 was constructed with PCR overlapping primers (BoNT/B LC), cloned in a pQE30 UA vector, and expressed in an E. coli M15 host system. Recombinant protein production was optimized at 0.5 mM IPTG final concentration, 4 h post induction, resulting in a maximum yield of recombinant proteins. The immunogenic nature of the recombinant BoNT/B LC protein was evaluated by ELISA. Antibodies were raised in BALB/c mice using various adjuvants. A significant rise in antibody titer (p<0.05) was observed in the Alum group, followed by the Titermax Classic group, Freund's adjuvant, and the Titermax Gold group. These developed high-titer antibodies may prove useful for the detection of botulinum neurotoxins in food and clinical samples.