1.Development of a Disperse Dye Immunoassay Technique for Detection of Antibodies against Neospora caninum in Cattle.
Fatemeh SELAHI ; Mehdi NAMAVARI ; Mohammad Hossein HOSSEINI ; Maryam MANSOURIAN ; Yahya TAHAMTAN
The Korean Journal of Parasitology 2013;51(1):129-132
In this study a disperse dye immunoassay method was standardized and evaluated for detection of antibodies against Neospora caninum in cattle. Sera from 150 cattle with a recent history of abortion were collected and tested by commercial ELISA kit and a standardized in-house dye immunoassay system. The positivity rate for the sera used in this study was 34.6% for the disperse dye immunoassay (DDIA) compared to 32% obtained by ELISA kit. This study showed no significant difference between DDIA and ELISA. The results indicated that the DDIA provide an economic, simple, rapid and robust test for detection of N. caninum infection in cattle.
Animals
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Antibodies, Protozoan/*blood
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Cattle
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Cattle Diseases/*diagnosis/parasitology
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Coccidiosis/diagnosis/parasitology/*veterinary
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Diagnostic Tests, Routine/*methods
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Female
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Immunoassay/methods
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Neospora/*immunology
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Staining and Labeling/methods
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Veterinary Medicine/*methods
2.Prevalence of serum allergen-specific immunoglobulin E for canine atopic dermatitis in Korea.
Hyo Mi JANG ; Gwi Seon YEO ; Ji Hyun KIM ; Cheol Yong HWANG ; Jae Eun HYUN ; Soon Shin KIM ; Yang Ho KANG ; Dong In JUNG
Journal of Biomedical Research 2014;15(4):162-169
Canine atopic dermatitis (CAD) is an allergic skin disease with characteristic clinical features associated with immunoglobulin E (IgE) antibodies. Identification of the causative allergens is the diagnostic goal, which is essential to treat and manage CAD patients. CAD is commonly associated with environmental allergens surrounding the patients. For this reason, it is important for diagnostic tests to select allergens that are related to the environment of each country and each province. There are two main allergen-specific tests, serological IgE test (SAT) and intradermal skin test (IDT). SAT did not show direct cutaneous reaction but did show serological reaction against allergens. However, SAT is simpler and more convenient than IDT in small animal practice. In this study, we selected domestically prevalent allergens for SAT, including 60 food allergens and 60 inhalant allergens, and tested eight dogs tentatively diagnosed with CAD based on Favrot's criteria. Furthermore, IDT was performed on four dogs from the SAT group for comparison of SAT and IDT, and the results were very similar. In SAT, four types of mites (Bloomia tropicalis, Glycophagus domesticus, Euroglyphus maynei, and mite mixture 1 Korea; house dust mites), four types of molds (Botrytis cinerea, Alternaria alternata, mold fungi mixture 11, mold fungi mixture), and one type of pollen (tree pollen mix 3 Korea) induced a reaction in more than half of dogs tested. In IDT, all four dogs reacted positively to Dermatophagoides farinae, and three reacted positively to Dermatophagoides pteronyssinus and house dust. The mean agreement rate between SAT and IDT in this study was 76.3%. This is the first trial to apply local allergens for SAT in Korean veterinary medicine, and it might play an important role for diagnoses and management of animal allergic diseases.
Allergens
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Alternaria
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Animals
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Antibodies
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Dermatitis, Atopic*
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Dermatophagoides farinae
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Dermatophagoides pteronyssinus
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Diagnosis
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Diagnostic Tests, Routine
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Dogs
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Dust
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Fungi
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Humans
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Immunoglobulin E*
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Immunoglobulins*
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Korea
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Mites
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Pollen
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Prevalence*
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Pyroglyphidae
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Serologic Tests
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Skin Diseases
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Skin Tests
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Veterinary Medicine
3.Development and clinical evaluation of a rapid diagnostic kit for feline leukemia virus infection.
Won Shik KIM ; Chom Kyu CHONG ; Hak Yong KIM ; Gyu Cheol LEE ; Wooseog JEONG ; Dong Jun AN ; Hye Young JEOUNG ; Jae In LEE ; Young Ki LEE
Journal of Veterinary Science 2014;15(1):91-97
Feline leukemia virus (FeLV) causes a range of neoplastic and degenerative diseases in cats. To obtain a more sensitive and convenient diagnosis of the disease, we prepared monoclonal antibodies specific for the FeLV p27 to develop a rapid diagnostic test with enhanced sensitivity and specificity. Among these antibodies, we identified two clones (hybridomas 8F8B5 and 8G7D1) that specifically bound to FeLV and were very suitable for a diagnostic kit. The affinity constants for 8F8B5 and 8G7D1 were 0.35 x 10(9) and 0.86 x 10(9), respectively. To investigate the diagnostic abilities of the rapid kit using these antibodies, we performed several clinical studies. Assessment of analytical sensitivity revealed that the detection threshold of the rapid diagnostic test was 2 ng/mL for recombinant p27 and 12.5 x 10(4) IU/mL for FeLV. When evaluating 252 cat sera samples, the kit was found to have a kappa value of 0.88 compared to polymerase chain reaction (PCR), indicating a significant correlation between data from the rapid diagnostic test and PCR. Sensitivity and specificity of the kit were 95.2% (20/21) and 98.5% (257/261), respectively. Our results demonstrated that the rapid diagnostic test would be a suitable diagnostic tool for the rapid detection of FeLV infection in cats.
Animals
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Antibodies, Monoclonal/blood
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Cats
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Diagnostic Tests, Routine/*veterinary
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Female
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Gene Products, gag/*blood
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Leukemia Virus, Feline/immunology/*isolation & purification
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Leukemia, Feline/*diagnosis
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Mice, Inbred BALB C
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Sensitivity and Specificity
4.Quantitative real-time polymerase chain reaction for detecting Mycoplasma hyosynoviae and Mycoplasma hyorhinis in pen-based oral, tonsillar, and nasal fluids.
Joao Carlos GOMES NETO ; Leslie BOWER ; Barbara Z ERICKSON ; Chong WANG ; Matthew RAYMOND ; Erin L STRAIT
Journal of Veterinary Science 2015;16(2):195-201
Mycoplasma (M.) hyorhinis and M. hyosynoviae are pathogens known to cause disease in pigs post-weaning. Due to their fastidious nature, there is increased need for culture-independent diagnostic platforms to detect these microorganisms. Therefore, this study was performed to develop and optimize quantitative real-time PCR (qPCR) assays to rapidly detect M. hyorhinis and M. hyosynoviae in pen-based oral fluids as well as nasal and tonsillar fluids as proxies for samples used in swine herd surveillance. Two methods of genomic DNA extraction, automated versus manual, were used to compare diagnostic test performance. A wean-to-finish longitudinal study was also carried out to demonstrate the reproducibility of using pen-based oral fluids. Overall, pen-based oral and tonsillar fluids were more likely to be positive for both types of bacteria whereas only M. hyorhinis was detected in nasal fluids. DNA extraction protocols were shown to significantly influence test result. Although the initial detection time somewhat differed, both organisms were repeatedly detected in the longitudinal study. Overall, this study evaluated two qPCR methods for rapid and specific detection of either mycoplasma. Results from the present investigation can serve as a foundation for future studies to determine the prevalence of the two microorganisms, environmental load, and effectiveness of veterinary interventions for infection control.
Animals
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Diagnostic Tests, Routine/methods/*veterinary
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Female
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Longitudinal Studies
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Mouth/microbiology
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Mycoplasma Infections/diagnosis/microbiology/*veterinary
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Mycoplasma hyorhinis/*isolation & purification
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Mycoplasma hyosynoviae/*isolation & purification
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Nose/microbiology
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Palatine Tonsil/microbiology
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Real-Time Polymerase Chain Reaction/*veterinary
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Reproducibility of Results
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Swine
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Swine Diseases/*diagnosis/microbiology