• Pediatric Infection and Vaccine
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• v.19 no.2
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• pp.55-60
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• 2012

Purpose: We conducted the immunoassay of pertussis according to ages, in order to evaluate protective immunity against pertussis in Korean populations. Methods: Healthy subjects were enrolled at four university hospitals in Korea. The subjects were grouped as seven age groups (every 10 years). Antibodies against pertussis toxin (PT) in sera were measured by enzyme linked immunosorbent assay (ELISA) kits. Geometric mean concentrations (GMC) of antibodies and the ratios of the subjects with seroprotective antibody levels were determined. The subjects with antibody titers ${\geq}24.0EU/mL$ were considered to seroprotective as the manufacturer's protocol. Results: Total 1,605 subjects (age: 2 months-65 years) participated in this study, and their GMC was $56.16{\pm}50.54EU/mL$. Among seven age groups, age group <11 year showed the highest GMC ($64.78{\pm}53.24EU/mL$) (P<0.001). In the analysis of the ratios of the subjects with seroprotective antibody titers, 68.2% of the subjects were proven to seroprotective, and age group <11 year also showed the highest ratio (76.5%) (P<0.001). Conclusions: We found that adolescences or adults (age group ${\geq}11$ year) showed lower levels of antibody against pertussis and lower ratio of the subjects with seroprotective antibody titers than children (age group <11 year).

• Journal of Veterinary Clinics
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• v.31 no.4
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• pp.267-271
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• 2014

Accurate determination of in vivo oocyte maturation is particularly critical for dog cloning compared to other assisted reproductive technologies because oocytes in metaphase II stage have to be recovered in order to undergo somatic cell nuclear transfer right after its recovery. The aim of present study was to evaluate the reliability and to set a reference range of a chemiluminescence enzyme immunoassay (CLEIA) compared to radioimmunoassay (RIA) method to retrieve in vivo matured oocytes. Serum progesterone concentration during proestrus and estrus was analyzed by RIA and CLEIA to determine ovulation day (Day 0). On Day 3, in vivo oocytes were recovered surgically and evaluated microscopically maturation status after staining nucleus with bisbenzimidazole dye. Mean progesterone concentration by CLEIA ($7.64{\pm}0.06ng/ml$) was significantly higher than by RIA ($6.46{\pm}0.04ng/ml$, P < 0.0001). It was not different between CLEIA ($10.01{\pm}0.34ng/ml$) and RIA values ($7.91{\pm}0.14ng/ml$, P < 0.05) on Day 0, but significantly higher CLEIA level on Day -1 and Day 1 ($6.41{\pm}0.15$ and $14.25{\pm}0.44ng/ml$) was assessed compared to RIA ($4.95{\pm}0.10$ and $11.29{\pm}0.34ng/ml$). However, with both methods, progesterone level was significantly increased from Day -1 to Day 2. To determine oocyte maturation with CLEIA method, a wider and higher reference range has to be considered.

• Clinical and Experimental Pediatrics
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• v.52 no.1
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• pp.56-60
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• 2009

Purpose : This study aimed to test whether rotavirus-associated necrotizing enterocolitis (RV+NEC) produced different clinical findings or outcomes from those of non-rotavirus necrotizing enterocolitis (RV-NEC). Methods : Eight patients from the RV+NEC group and 22 patients from the RV-NEC group diagnosed with modified Bell stage II or higher NEC were selected for this study. Fecal specimens from all infants were tested for rotavirus infection using a monoclonal antibody-based enzyme immunoassay (EIA). Clinical, radiographic, and clinical outcome data were analyzed retrospectively. Results : RV+NEC infants had a significantly higher birth weight and were born at a significantly higher gestational age ($33.5{\pm}3.3$ weeks vs. $29.3{\pm}4.4$ weeks; P=0.01). There were no differences in the occurrence of thrombocytopenia, mural gas, and pneumoperitoneum between the 2 groups. However, portal vein gas was more common in the RV+NEC group (88% vs. 9%; P<0.01). Neither the incidence of Bell stage III (or higher) NEC nor surgical intervention differed between the two groups. The number of complications and mortality rates were also similar. Conclusion : Rotavirus-associated NEC occurs in infants with a higher birth weight and those born at a greater gestational age. However, the severity of the condition and the resulting outcomes did not differ from those for infants affected by non-rotavirus NEC.

• Proceedings of the IEEK Conference
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• 2005.11a
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• pp.755-758
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• 2005

This paper describes the novel immunoassay sensing system for a portable clinical diagnosis system. It consists of a bead cage reactor and a CMOS integrated biosensor. It showed the simple and easy antibody coating method on beads by flow-through avidin biotin complex technology in a microfluidic device. It showed just 90 nL sample consumption and good result for the application of alpha feto protein. The bead cage reactor has the role of the antibody coating, antigen binding and enzyme linking for the electrochemical sensing method. The CMOS biosensor consists of ISFET (ion selective field effect transistor) biosensor and temperature sensor for detecting pH that is the byproduct of enzyme reaction. The sensitivity is 8 $kHz/^{\circ}C$ in a temperature sensor and 33 mV/pH in a pH sensor. After filling the 15 um polystyrene beads in bead cage, antibody flowed and reacted to beads. Subsequently, the biotinylated antigen flowed and bound to the antibody and GOD (glucose oxidase)-avidin conjugate flowed and reacted to the biotin of the biotinylated antigen. After this reaction process, glucose solution flowed and reacted to the GOD on beads. The hydrogen was generated by glucose-GOD reaction. And it was detected by the pH sensor.

• Journal of Physiology & Pathology in Korean Medicine
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• v.18 no.6
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• pp.1694-1698
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• 2004

We hope to evaluate the effects of Sabaek-san for the bronchial asthma using assesment on the metrix metalloproteinase-9(MMP-9) after Sabaek-san was intravenously administered OVA-sensitized and -challenged mice. Seventy-two female mice, 8-10 weeks of age and free of murine specific pathogens, were used. Of the seventy-two mice, twenty-four mice were not sensitized and forty-eight mice were sensitized by intraperitoneal injection of OVA. Of the sensitized mice, twenty-four mice didn't administrate Sabaek-san and twenty-four administrated Sabaek-san. Mice were sensitized on days 1 and 14 by intraperitoneal injection of 20 fig OVA. On days 21, 22 and 23 after the initial sensitization, the mice were challenged for 30 minutes with an aerosol of 1% OVA in saline. Sabaek-san administered 200㎎/㎏ in the tail of the mouse, one time per day, for 7 days, beginning 14 days after first sensitization. Bronchoalveolar lavage was performed 72 hours after the last challenge, and total cell numbers in the BAL fluid were count. Also, level of MMP-9 in the BAL fluid were measured by Enzyme immunoassays and Western blot analysis. Enzyme immunoassay revealed that MMP-9 levels in the BAL fluids significantly increased 72 h after OVA inhalation compared with levels in the control group. After administration of the Sabaek-san, the levels of the MMP-9 in BAL fluids 72 h after OVA inhalation reduced dramatically. Western blot analysis revealed that MMP-9 levels increased in the all mice which were challenge with OVA without administered Sabaek-san compared the normal mouse. However, in the groups of the administered Sabaek-san, the MMP-9 level markedly decreased. Sabaek-san might be effect the treatment of the bronchial asthma as a inhibition of the MMP-9.

• Journal of Microbiology
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• v.42 no.3
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• pp.211-215
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• 2004

Toxoplasmosis has been well known as an important human infection to consider especially in pregnant women. Although many serologic methods are available, the diagnosis of toxoplasmosis can be extremely difficult. The presence of increased levels of Toxoplasma-specific IgG antibodies indicates an infection, but it does not differentiate between a recent and past infection. The purpose of our study was to compare the performance of the ELISA T. gondii IgG/IgM test, a widely used enzyme-linked immunosorbent assay, to the ELISA IgG avidity method. One hundred and four serum samples (from 38 males and 66 females) were tested and evaluated from symptomatic patients (chorioretinitis, lymphadenopathy), and from women in their first trimester of pregnancy who were suspected of having toxoplasmosis, The high IgG avidity and ELISA IgG antibody levels were in agreement for 51 of the specimens (49.0％). Thirty-eight discrepant (borderline) results from the IgG avidity method were positive for IgM (3 specimens) and IgG (37 specimens). Interestingly, out of the eight serum samples that were positive for both IgG and IgM antibodies, two samples were low IgG avidity, and three samples were borderline. There was no statistically significant relation observed between the results of the IgG avidity method and the ELISA IgG test, and the IgG avidity method and ELISA IgM test (X$^2$=1.987; p=0.370 and X$^2$=2.152; p=0.341, respectively). The IgG avidity method was considered easy to perform and an acceptable approach for the differentiation of discrepant results (recent/chronic) and for the current detection of T. gondii antibodies. We concluded that the determination of IgG avidity is a helpful tool for the diagnosis of the ocular form of toxoplasmosis and it is a safe method for screening this disease in the first trimester of pregnancy.

• Clinical and Experimental Pediatrics
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• v.49 no.11
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• pp.1125-1139
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• 2006

In 1991, the Ministry of Health & Social affairs adopted a nationwide service program for neonatal screening of phenylketonuria, galactosemia, maple syrup urine disease, homocystinuria, histidinemia & congenital hypothyroidism for newborns delivered from low class pregnant women registered in health centers. Government decreased the test items from six to two, PKU & congenital hypothyroidism to increase test numbers with same budget from 1995. Government decided to test PKU & hypothyroidism for all newborns from 1997. 78 laboratories wanted to participate for neonatal screening test in 1999. Government didn't decide laboratory center for a certain district and placed responsibility on free competition. Government are planning to test 573,000 newborns from 1998, Government decided to screen 6 items PKU, congenital hypothyroidism, maple syrup urine disese, homocystinuria, galactosemia and congenital adrenal hyperplasia from 2006. 17 laboratores are participating now. The cost of screening test is supported by both the federal government and local government on a 40-60 basis. In case a patient with an inherited metabolic disease is diagnosed by screening of government program, special milk is provided at government's expense. Interlaboratory quality control was started 6 times a year from 1994. According to the government project, 3,707,773 newborns were screened. 86 PKU, 718 congenital hypothyroidism were detected. So incidence of PKU is 1/43,114 and congenital hypothyroidism is 1/4,612. Maeil dairy company produced new special formula for PKU, MMA and PA, MSUD, urea cycle disorder, homocystinuria, isovaleric acidemia from Oct. 1999. The cost benefit of performing screening procedures coupled with treatment has been estimated to be as high as 1.77 times in PKU, 11.11 times in congenital hypothyroidism than cost without screening. We are trying to increase the budget to test all newborns for Tandem mass sereening & Wilson disease from 2008. Now it is a very important problem to decrease laboratory numbers of neonatal screening in Korea. So we are considering 4-5 central laboratories which cover all newborns and are equipped with tandem mass spectrometer & enzyme immunoassay for TSH, 17OHP & enzyme colorimetric assay for galactose.

• The Journal of Pediatrics of Korean Medicine
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• v.20 no.1
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• pp.181-194
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• 2006

Objective : Author evaluate the effects of Citri Reticulatae Viride Pericarpium for the bronchial asthma using assesment of the vascular endothelial growth factor after Citri Reticulatae Viride Peicarpium was intravenously administered Ovalbumine(OVA)-sensitized and -challenged mice. Material and Methods : Twenty-four female mice, 8-10 weeks old and free of murine specific pathogens, were sensitized by intraperitoneal injection of OVA. Of the sensitized mice, ten mice didn't administrate Citri Reticulatae Viride Pericarpium and the other ten mice administrated Citri Reticulatae Viride Pericarpium. Mice were sensitized on the first and fourteen days introspectional injection of 20 ${\mu}g$ OVA. After$21^{st}\;22^{th}\;and\;23^{rd}$, the initial sensitization, the mice were challenged for 30 minutes with an aerosol of 1% OVA in saline. Citri Reticulatae Viride Pericarpium administered 200mg/kg in the tail of the mouse, one a day, for 7 days and beginning 14 days after first sensitization. Bronchoalveolar lavage was performed 72 hours after the last challenge, and total cell numbers in the BAL fluid were count. Also, lever of VEGF in the BAL fluid was measured by Enzyme immunoassays and Western blot analysis. Results: Total cell numbers in BAL fluid were significantly greater than from 72 hrs after OVA inhalation compared with cell numbers in the control group. However, there was no difference of the total cell numbers between OVA-challenge groups without Citri Reticulatae Viride Pericarpium, and OVA-challenge with Citri Reticulatae Viride Pericarpium. Enzyme immunoassay revealed that VEGF levels in the BAL fluids were significantly increased 72 hrs after OVA inhalation compared with levels in the control group. After administration of the Citri Reticulatae Viride Pericarpiu, the levels of the VEGF in BAL fluids 72 hrs after OVA inhalation reduced dramatically. Western blot analysis revealed that VEGF protein levels were increased in the all mice which were challenge with OVA, without administered Citri Reticulatae Viride Pericarpium, compared the normal mouse. However, in the groups of the administered Citri Reticulatae Viride Pericarpium, the VEGF protein level markedly decreased. Conclusion: Citri Reticulatae Viride Pericarpium might affect the treatment of the bronchial ashma as a inhibition of the VEGF.

• Journal of The Korean Society of Inherited Metabolic disease
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• v.14 no.1
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• pp.1-9
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• 2014

Many inborn errors of metabolism can be completely cured with early detection and early treatment. This is why neonatal screening on inborn errors of metabolism is implemented worldwide. In 1991, the Ministry of Health & Social affairs adopted a nationwide service program for neonatal screening of phenylketonuria, galactosemia, maple syrup urine disease, homocystinuria, histidinemia and congenital hypothyroidism for newborns delivered from low class pregnant women registered in health centers. Government decreased the test items from six to two, PKU and congenital hypothyroidism to increase test numbers with same budget from 1995. 78 laboratories wanted to participate for neonatal screening test in 1999. Government decided to screen six items of PKU, congenital hypothyroidism, maple syrup urine disease, homocystinuria, galactosemia and congenital adrenal hyperplasia from 2006. In 2014, thirteen laboratories are participating. Inter laboratory quality control was started 6 times a year from 1994. In case a patient with an inherited metabolic disease is diagnosed by screening of government program, special milk is provided at government's expense. According to the government project, from 1997 to 2013, 7,080,569 newborns were screened. 144 PKU, 2.451 congenital hypothyroidism were detected. So incidence of PKU is 1/49,170 and congenital hypothyroidism is 1/2,888. The cost benefit of performing screening procedures coupled with treatment has been estimated to be as high as 1.77 times in PKU, 11.11 times in congenital hypothyroidism than cost without screening. By January 2007, many European countries had expanded of their newborn screening programs by inclusion of Tandem mass spectrometry. We are trying to increase the budget to test all newborns for Tandem mass spectrometry from 2016. We are considering four to five central laboratories which cover all newborns and are equipped with tandem mass spectrometer & enzyme immunoassay for TSH, 17OHP & enzyme colorimetric assay for galactose. And I hope to expand test including Wilson disease screening test and lysosomal storage diseases.

• 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.