• ANNALS OF ANIMAL RESOURCE SCIENCES
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• v.28 no.2
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• pp.78-96
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• 2017

Foot-and-mouth disease (FMD) is a infection that can easily spread when it occurs and causes serious economic damage because of the existence of multiple serotypes of the virus and extreme contagiousness. The most effective method in preventing the transmission of FMD virus (FMDV) is the culling of livestock and additional vaccination in the other areas depending on the spreading rate and situation. Diagnostic methods are utilized not only for the definite diagnosis of FMD but also for identification of serotype, and confirmation of antibody production after vaccination. Although many methods have been developed to diagnose, they are not still enough to detect accurately the disease in a short time. Therefore, it has been needed new diagnostic methods improved from existing methods. Previous methods were based on the enzyme-linked immunosorbent assay (ELISA) as a serological diagnostic method, or polymerase chain reaction (PCR), which is a molecular genetic method. The recent technology has been performing about the combination of both methods and how to make it faster, less costly, more sensitive and accurate way.

• Journal of Veterinary Science
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• v.23 no.3
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• pp.50.1-50.12
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• 2022

Background: There is an urgent need to find reliable and rapid bovine tuberculosis (bTB) diagnostics in response to the rising prevalence of bTB worldwide. Toll-like receptor 2 (TLR2) recognizes components of bTB and initiates antigen-presenting cells to mediate humoral immunity. Evaluating the affinity of antigens with TLR2 can form the basis of a new method for the diagnosis of bTB based on humoral immunity. Objectives: To develop a reliable and rapid strategy to improve diagnostic tools for bTB. Methods: In this study, we expressed and purified the sixteen bTB-specific recombinant proteins in Escherichia coli. The two antigenic proteins, MPT70 and MPT83, which were most valuable for serological diagnosis of bTB were screened. Molecular docking technology was used to analyze the affinity of MPT70, MPT83, dominant epitope peptide of MPT70 (M1), and dominant epitope peptide MPT83 (M2) with TLR2, combined with the detection results of enzyme-linked immunosorbent assay to evaluate the molecular docking effect. Results: The results showed that interaction surface Cα-atom root mean square deviation of proteins (M1, M2, MPT70, MPT83)-TLR2 protein are less than 2.5 A, showing a high affinity. It is verified by clinical serum samples that MPT70, MPT83, MPT70-MPT83 showed good diagnostic potential for the detection of anti-bTB IgG and M1, M2 can replace the whole protein as the detection antigen. Conclusions: Molecular docking to evaluate the affinity of bTB protein and TLR2 combined with ELISA provides new insights for the diagnosis of bTB.

• Research in Plant Disease
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• v.20 no.3
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• pp.173-181
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• 2014

The early and accurate detection of plant viruses is an essential component to control those. Because the globalization of trade by free trade agreement (FTA) and the rapid climate change promote the country-to-country transfer of viruses and their hosts and vectors, diagnosis of viral diseases is getting more important. Because symptoms of viral diseases are not distinct with great variety and are confused with those of abiotic stresses, symptomatic diagnosis may not be appropriate. From the last three decades, enzyme-linked immunosorbent assays (ELISAs), developed based on serological principle, have been widely used. However, ELISAs to detect plant viruses decrease due to some limitations such as availability of antibody for target virus, cost to produce antibody, requirement of large volume of sample, and time to complete ELISAs. Many advanced techniques allow overcoming demerits of ELISAs. Since the polymerase chain reaction (PCR) developed as a technique to amplify target DNA, PCR evolved to many variants with greater sensitivity than ELISAs. Many systems of plant virus detection are reviewed here, which includes immunological-based detection system, PCR techniques, and hybridization-based methods such as microarray. Some of techniques have been used in practical, while some are still under developing to get the level of confidence for actual use.

• Asian Pacific Journal of Cancer Prevention
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• v.14 no.7
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• pp.4295-4299
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• 2013

Background: Pretreatment serum p53 and epidermal growth factor receptor (EGFR) were assessed using enzyme-linked immunosorbent assay (ELISA) in patients with acute leukemia to analysis their roles in characterization of different subtypes of the disease. Materials and Methods: Serum samples from thirty two patients with acute myeloid leukemia (AML) and fourteen patients with acute lymphoid leukemia (ALL) were analysed, along with 24 from healthy individuals used as a control group. Results: The results demonstrated a significant increase of serum p53 and EGFR in patients with AML (p<0.0001) compared to the control group. Also, the results showed a significant increase of both markers in patients with ALL (p<0.05, p<0.0001 respectively). Sensitivities and specificities for these variables were 52% and 100% for p53, and 73.9%, 95.8% for EGFR. Serum p53 and EGFR could successfully differentiate between M4 and other AML subtypes, while these variables failed to discriminate among ALL subtypes. A positive significant correlation was noted between p53 and EGFR. Negative significant correlations were observed between these variables and both of hemoglobin (Hg) content and RBC count. Conclusions: Mutant p53 and EGFR are helpful serological markers for diagnosis of patients with AML or ALL and can aid in characterization of disease. Moreover, these markers may reflect carcinogenesis mechanisms.

• Korean Journal of Veterinary Research
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• v.42 no.4
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• pp.545-550
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• 2002

The raccoon dog (Nyctereutes procyonoides) may be infected by Dirofilaria immitis. However, there has been no report on dirofilarial infection in the raccoon dog in Korea. In this study, we report on D. immitis infection in two wild raccoon dogs captured in the Daejeon area. The two raccoon dogs were referred to the Veterinary Teaching Hospital, Chungnam National University for diagnosis of D. immitis infection. The modified Knott's test for the detection of blood D. immitis microfilariae was positive, and serological test (FASTest$^{(R)}$ HW Antigen ELISA kit, Diagnostik Mega Cor, Austria) for D. immitis was positive as well. Additionally, D. immitis microfilariae were differentiated from other microfilariae by using acid phrnphatase histochemical staining (Leucognost-SP$^{(R)}$kit, Diagncstica MERCK, Germany). The two raccoon dogs were necropsed and D. immitis infection was confirmed.

• Korean Journal of Veterinary Research
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• v.43 no.3
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• pp.471-476
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• 2003

Malignant catarrhal fever (MCF) is a systemic disease of ruminants caused by ovine herpesvirus 2 (OvHV-2). OvHV-2 is a gamma herpesvirus, which induces frequent latent infection and often difficult to detect its antigens and even specific nucleic acids because of its low viral copies in the infected tissues. Histopathology, serology and polymerase chain reaction (PCR) were compared for the diagnosis of MCF using 10 bison infected with OvHV-2. Histopathological diagnosis was performed using the criteria which was based upon the pathognomic lesions. Serological diagnosis was conducted using its serum with competitive ELISA for the detection of antibodies of OvHV-2. Also, the nest PCR was performed with peripheral blood leukocytes for the detection of OvHV-2-specific DNAs. Primers 556 and 775 were used for the primary amplification, and primers 556 and 555 were used for the secondary amplification. As the results, positive cases were 6 by histopahology, 9 by serology and 10 by PCR. As comparing with other diagnostic methods, PCR was found to be more sensitive than histopathology and serology. The recent development of molecular diagnostic assays has provided powerful tools for investigating how viruses survive in nature. Development of PCR specific for viruses has dramatically improved the accuracy of diagnosis of viruses in clinically infected animals. Furthermore, amplification of viral genomic material by nest PCR represents the most sensitive method for the detection of viruses and might be detected successfully even though very low viral DNA copies. So, it could be used as the first choice for the detection of viral DNAs with low copies such as the status of latent infection. However, it has also some limitation of application like as false negative results by PCR inhibitors and false positive results by contamination. The results of this study suggest that the use of molecular biological methods like PCR may increase the accuracy for the diagnosis of infectious diseases. However, in diagnostic laboratory, it is recommended that PCR assay must be conducted with other diagnostic methods for more reliable diagnosis.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.15 no.2
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• pp.85-90
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• 2012

Purpose: Commercial enzyme-linked immunosorbent assay (ELISA) kits have been considered less reliable for children than for adults. The aim of this study was to compare four ELISA kits and in-house immunoblotting based on the analysis of anti-H. pylori-IgG antibody reactivity. Methods: A total of 399 serum samples were collected at the GNU Hospital during 1998-1999. All sera were tested using ELISA and immunoblotting. Statistically significant differences were determined by the $x^2$ test. Results: The overall seropositivity rates using GAP IgG, Genedia IgG, HM-CAP, Pyloriset EIA-G, and immunoblotting were 13.0%, 25.1%, 18.3%, 15.8%, and 62.9%, respectively. Immunoblotting showed a higher seropositivity rate than did all four ELISA kits in all age groups. Genedia IgG had the highest seropositivity among the ELISA kits. The seropositivity rate for children aged 13 to 18 months was lowest, and that of children aged 15 years was highest (90.0%). The seropositivity rate for children aged 7 months to 5 years was significantly lower than that for children aged 6 to 15 years among the four ELISA kits (p<0.0001) and immunoblotting (p=0.02). Conclusion: Immunoblotting is the most sensitive test for detection of anti-Helicobacter pylori IgG antibodies among the serological tests in this study. These results emphasize the need for standardization when commercial ELISA tests are used in different nations or in young age groups. Immunoblotting could be a suitable noninvasive assay for serodiagnosis and seroepidemiologic study of H. pylori infection in Korean children.

• Journal of Veterinary Science
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• v.22 no.3
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• pp.29.1-29.6
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• 2021

West Nile virus (WNV), a neurotropic arbovirus, has been detected in mosquitos, birds, wildlife, horses, and humans in Malaysia, but limited information is available on WNV infection in Malaysian pigs. We tested 80 archived swine serum samples for the presence of WNV antibody and West Nile (WN) viral RNA using ID Screen West Nile Competition Multi-species enzyme-linked immunosorbent assay kits and WNV-specific primers in reverse transcription polymerase chain reaction assays, respectively. A WNV seroprevalence of 62.5% (50/80) at 95% confidence interval (51.6%-72.3%) was recorded, with a significantly higher seroprevalence among young pigs (weaner and grower) and pigs from south Malaysia. One sample was positive for Japanese encephalitis virus antibodies; WN viral RNA was not detected in any of the serum samples.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.12
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• pp.5265-5272
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• 2016

Objective: The present study was conducted to evaluate invasive and noninvasive diagnostic methods for detection of Helicobacter pylori (H. pylori) in patients admitted with dyspeptic complaints and to compare sensitivities and specificities. Method: Sets of four gastric biopsy specimens were obtained from a total of 126 patients included in the study. The presence of H. pylori was determined by invasive tests including culture, rapid urease test, polymerase chain reaction (PCR) and histopathology. Among noninvasive tests, urea breath test, serological tests and enzyme-linked immunosorbent assay (ELISA) were performed. Results: H. pylori was isolated in 79 (62.7%) gastric biopsy cultures, whereas positivity was concluded for 105 (83.3%) patients by rapid urease test, for 106 (84.1%) by PCR, for 110 (87.3%) by histopathology, for 119 (94.4%) by urea breath test, and for 107 (84.9%) by ELISA. In the present study, the culture findings and histopathological examination findings were accepted as gold standard. According to the gold standard, urea breath test had the highest sensitivity (96.5%) and the lowest specificity (30%), whereas culture and histopathology had the highest specificities (100%). Conclusion: The use of PCR invasively with gastric biopsy samples yielded parallel results with the gold standard. PCR can be recommended for routine use in the diagnosis of H. pylori.

• Journal of Yeungnam Medical Science
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• v.29 no.2
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• pp.89-95
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• 2012

Background: As Mycoplasma pneumoniae pneumonia has increased in Korea, its relevance to infants, toddlers, and adolescents has magnified as well as. However, it is difficult to perform the serological test and PCR test routinely for diagnosis in actual clinical practice. Thus, the authors conducted this study to help clinicians do presumptive diagnosis of Mycoplasma pneumoniae pneumonia using clinical, radiological, and hematological findings. Methods: The study population consisted of 224 children between 1 month and 14 years old, hospitalized for radiographically confirmed pneumonia. Patients were divided into two groups of 100 children with Mycoplasma pneumoniae pneumonia, as diagnosed using the ELISA method. Groups with negative result in Mycoplasma IgM antibody test were classified into the viral group (98 patients with respiratory virus) and the bacterial group (46 patients with the bacteria detected in the blood sputum culture or antibiotic treatment except macrolide improved the patient's condition). These groups were compared and analyzed using clinical, hematological, and radiographic differences and scoring system. Results: Clinical, hematological, and radiographic characteristics of Mycoplasma pneumoniae pneumonia have shown the intermediate level results between bacterial pneumonia and viral pneumonia. In terms of scoring system, the mean score of Mycoplasma pneumoniae pneumonia was 4.23, which was the intermediate level between bacterial pneumonia (mean score=6.67) and viral pneumonia (mean score=1.48). Conclusion: Results suggest that the combination of the scoring system information can increase the accuracy in the diagnosis even if they may have difficulties on diagnosis, because clinical manifestations, hematological, and radiographic findings are nonspecific.