• Title/Summary/Keyword: Diagnostic kit

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Comparison of the non-invasive diagnostic methods, stool antigen test and PCR assay, for Helicobacter felis detection in dogs

  • Hong, Sunhwa;Lee, Hak-Yong;Kim, Tae-Wan;Kim, Okjin
    • Korean Journal of Veterinary Service
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    • v.38 no.1
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    • pp.37-42
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    • 2015
  • The aim of the present study was to compare the non-invasive methods for the diagnosis of H. felis with HpSA kit-based detection method and H. felis-specific PCR assay with dog's stool samples without sacrifice. Male Beagle dogs (n=6) were infected with H. felis ATCC 49179 ($1.0{\times}10^9CFU/dog$) by intra-gastric inoculation two times at 3-day intervals, and the stool specimens of dogs were collected 1, 3, 5, 7, 14, 21 days after infection to submit to HpSA test and H. felis-specific PCR. As the results, the sensitivity of the HpSA and the PCR analysis was 50.0%, 83.3% respectively. Although HpSA test is less sensitive, it could be used for rapid, cheap and easy screening assay for H. felis infection in dog and cats. We suggest that the H. pylori stool antigen kit, HpSA, is useful and effective for monitoring H. felis infection. If HpSA test would be made with H. felis antibodies in the future, its sensitivity could be increased. Also, PCR assay could be successfully used to detect the H. felis in stools. Applying the H. pylori stool antigen kit and PCR assay may be the recommended non-invasive strategy to identify H. felis in dog and cats.

Stabilization of Dry Immuno-Diagnostic Kit with Immobilized Antibody and Determination of Its Shelf-Life (항체가 건조상태로 고정된 면역진단키트의 안정화와 Shelf-Life 결정)

  • 이창우;조정환;육순학;권오협;박영남;박세환
    • KSBB Journal
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    • v.13 no.5
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    • pp.502-510
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    • 1998
  • Stabilization of antibody, which is specific to Salmonella typhimurium antigens, present in dry states on membranes was accomplished, and its shelf-life, i.e., duration for maintaining minimum 90% of the initial activity, under optimal conditions was determined. To prepare two major components of an immuno-strip, the antibody was not only immobilized on nitrocellulose membrane surfaces but also placed within the pores of glass fiber membrane after conjugating it with old colloids as signal generator. Among potential stabilizers of the immuno-components, a disaccharide, trehalose, showed a significant protection effect of immunoglobulin structure from thermal energy. Optimal concentrations of trehalose for the respective component were significantly different (8-fold higher for the antibody-gold conjugate than for the immobilized antibody), which probably resulted from distinct densities and configurations of antibody present on the membranes. An additional requirement for the gold conjugate was freeze-drying of this substance such that the conjugate can be readily resolubilized upon contact with aqueous medium. By using the components prepared under optimal conditions, immuno-strips were constructed and exposed to thermal energy. Signals with less than 10% decrease in the intensity were maintained for approximately 21 days at 60$^{\circ}C$. Compared to previous reports, this result represented a 2-year shelf-life at room temperature. it was, however, two times longer if determined from thermal acceleration tests based on the theory of inactivation rate of protein. Such discrepancy between the two estimates could be mainly attributed to errors in accurately controlling temperatures and also to changes in the physical properties of membranes due to a high thermal energy.

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Performance comparison and evaluation of interferon-gamma assay kit for bovine tuberculosis diagnosis (소 결핵 진단을 위한 인터페론감마 검사 키트의 성능 비교 평가)

  • Hong, Leegon;Choi, Woojae;Ro, Younghye;Ahn, Sunmin;Kim, Eunkyung;Choe, Eunhee;Kim, Danil
    • Korean Journal of Veterinary Service
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    • v.43 no.4
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    • pp.201-209
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    • 2020
  • In Korea, bovine tuberculosis (bTB) is a representative zoonotic disease that causes considerable economic loss. In determining the positive bTB, the ELISA method for examining the amount of interferon-gamma (IFN-γ) is included in Korea's diagnostic standard method. Recently, commercially available BIONOTE TB-Feron ELISA Plus (TB-Feron Plus) that detects IFN-γ has been introduced. However, since the scientific basis for the performance is limited, we evaluated performance by comparing it with the results of another IFN-γ ELISA assay kit (BOVIGAM®) certified by Office International des Epizooties. In our research, 42 positive blood samples preliminarily tested with a tuberculin skin test and/or BOVIGAM® and 54 negative blood samples collected from three bTB free farms were subjected to IFN-γ assay using the TB-Feron Plus and the BOVIGAM®, respectively. The result shows that the sensitivity, specificity and accuracy were 81.0% (34/42), 100% (54/54), 91.7% (88/96) in TB-Feron Plus kit and 78.6% (33/42), 100% (54/54), 90.6% (87/96) in BOVIGAM® kit, respectively. Moreover, the overall accordance percentage of the two kits was 99.0% (95/96) and there was almost perfect agreement between two assays (Kappa=0.977, P<0.0001). Furthermore, additional studies confirmed that elevated lymphocyte numbers in blood did not interfere with the results of the TB-Feron Plus kit. And, delayed time from sampling to culture decreased the optical density (OD) value. Therefore, we concluded that the TB-Feron Plus kit was not inferior to BOVIGAM® in performance. High lymphocyte numbers in blood did not impact on TB-Feron Plus results, while delayed time before culture interfered with OD value.

A study on production of early pregnancy diagnostic kit in cattle 1. Production of polyclonal antibody to progesterone and removal of anti-bovine serum albumin antisera (소의 조기임신진단 kit의 개발 1. Progesterone의 항체생산(抗體生産) 및 항(抗) BSA항체(抗體)의 제거)

  • Kang, Chung-boo;Lee, Hyo-jong;Choe, Sang-yong
    • Korean Journal of Veterinary Research
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    • v.31 no.2
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    • pp.217-222
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    • 1991
  • Most progesterone enzyme immunoassays(EIA) are used liquid phase double-antibody separation. These methods consume considerable time and reagents because of the requirements for several washing and centrifugation steps involving the reactants. Because of there several problems, we were prompted to develop an effective EIA system by the use of higher titer of progesterone antiserum free of anti-bovine serum albumin antibodies (anti-BSA). The results obtained were as follows. 1. The antibody of progesterone antiserum was high as $1.5{\times}10^5$. 2. Percent activity bound of progesterone antiserum was about 77 at a dilution to $5{\times}10^3$ times. 3. Progesterone antiserum was contained a large amount of anti-BSA antibodies. 4. The anti-BSA was completely absorbed by using of polymerised BSA. 5. The molecular weight of albumin polymer (polymerised BSA) obtained by using 2.5% glut. araldehyde was $5{\times}10^5$.

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A Study on Developing the Teachers' Guide Book for Diagnosis and Prescription of Students' Mathematical Errors (수학과 오류의 진단과 처방에 관한 교사용 자료 개발 연구)

  • 김수미
    • School Mathematics
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    • v.5 no.2
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    • pp.209-221
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    • 2003
  • This study focuses on the necessity of developing the material for teachers who are involved in diagnosing and prescribing students' mathematical errors. And it also intends to stimulate the related research of this area. For this, it tries to suggest the fundamental components-(1)types and frequencies of errors, (2) diagnostic test kit, (3)causes of errors, (4)ideas for prevention, (5)ideas for correction, (6)practice for settlement, and (7) performance test kit and frame of the teaching guide book for the teachers according to the general procedure of diagnosis and prescription. Finally it provides the concrete research areas for the future study.

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Analysis of prevalence and risk factors of calf diarrhea using rapid diagnosis kit (신속 진단 킷트를 활용한 송아지 설사병의 유병률과 위험요인 분석)

  • Taemook Park;Gil Jae Cho;Young Jin Yang;Il-Sun Ryu
    • Korean Journal of Veterinary Service
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    • v.46 no.1
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    • pp.47-58
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    • 2023
  • Between February 2020 and September 2021, 378 calves with diarrhea were investigated across 96 cattle breeding farms in Korea, using a rapid diagnostic kit. The study examined the infection rates of major pathogens causing diarrhea in calves, which were categorized by season, age, birth month, and region. Additionally, logistic regression analysis was conducted to investigate the factors affecting the infection rate. The study found that the five representative pathogens causing calf diarrhea exhibited differences in infection rates based on season, region, age, and birth month. Bovine rotavirus, bovine coronavirus, Cryptosporidium, and Giardia commonly exhibited varying risks of infection based on season and age. Furthermore, in addition to these risk factors, bovine rotavirus and Cryptosporidium were found to impact the infection risk of each pathogen by region, while Giardia was found to be affected by birth month.

Diagnostic Device Model for Insecticide susceptibilities of Beet Armyworm, Spodoptera exigua(Hubner) (파밤나방(Spodoptera exigua (Hiibner)) 살충제 감수성 진단장치모형)

  • 김용균;이준익;강성영;한상찬
    • Korean journal of applied entomology
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    • v.38 no.1
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    • pp.53-57
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    • 1999
  • Simple diagnostic kits for monitoring insecticide susceptibility of beet armyworm, Spodoptera exigua (Hiibner) were developed and applied to the field populations. The operation of the kits was based on the correlations between enzyme activities of esterase (EST) and acetylcholinesterase (AChE) and the insecticide susceptibilities. Four different kinds of diagnostic kits (ED, EM, AD, and AM) were designed and classified by diagnostic enzymes (E for esterases and A for acetylcholinesterase) and inhibitors (D for dichlorvos and M for monocrotophos). Diagnostic inhibitor concentrations were 1 mM for ED, 10 mM for EM, 100 mM for AD, and 100 mM for AM. Resistant larvae which were not inhibited by the diagnostic amounts of insecticides developed positive staining (red color), but susceptible~ s howed negative (no color). An insect was used for both EST and AChE diagnostic kits, but different in their samples: hemolymph for EST and the head for AChE. These four diagnostic kits were applied to 1 1 different populations which showed variations of insecticide susceptibilities. Four kits were different in the capability discriminating the insecticide susceptibilites according to insecticides: ED to bifenthrin, AD to methomyl, and ED and AM to chlorpyrifos-methyl. These diagnostic devices can be used for insecticide-resistance management program for this insect pest. It also provide a technical guide to insect pest management for farmers, directors, and researchers.

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Immunohistochemical Expression of Receptor Tyrosine Kinase (RTK) in Canine Brain Tumors

  • Jung, Hae-Won;Song, Joong-Hyun;Yu, Do-Hyeon;An, Su-Jin;Sur, Jung-Hyang;Kim, Young Joo;Han, Donghyun;Jung, Dong-In
    • Journal of Veterinary Clinics
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    • v.36 no.6
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    • pp.319-324
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    • 2019
  • Receptor tyrosine kinases (RTK) are major promising targets in anticancer therapy in human and veterinary medicine. Using immunohistochemistry method, we evaluated the expressionof five types RTK (PDGFR-α, PDGFR-β, VEGFR 2, c-Kit and Abl) in the six canine brain tumor samples (2 meningioma, 2 astrocytoma, 1 ependymoma and 1 choroid plexus papilloma). A total of five samples expressed PDGFR-β (5/6), one sample, the choroid plexus papilloma, expressed c-Kit (1/6), and a total of two samples expressed Abl (2/6). None of the samples showed expression of PDGFR-α and VEGFR 2. We demonstrate that a significant portion of canine brain tumors express tyrosine receptors for growth factors and show that these receptors generally localize to tumor cell membranes and the cytoplasm. Evaluation of immunohistochemical expression for the RTKs PDGFR-β, c-Kit, and Alb in canine brain samples reveals an interesting potential for molecular targeting by TKIs in therapeutic studies of canine brain tumors, and more studies will be needed to assess the interactions and efficacy of these RTKs and TKIs. Based on these results, we have some evidence for novel chemotherapeutic trials using TKIs for canine nervous tumors.

Ebola Hemorrhagic Fever Outbreaks: Diagnosis for Effective Epidemic Disease Management and Control (에볼라 출혈열 발병 : 효과적인 전염병 관리 및 통제를 위한 진단)

  • Kang, Boram;Kim, Hyojin;Macoy, Donah Mary;Kim, Min Gab
    • Microbiology and Biotechnology Letters
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    • v.45 no.2
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    • pp.87-92
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    • 2017
  • The first Ebola hemorrhagic fever outbreak occurred in the Democratic Republic of Congo and Sudan in 1976 and then emerged in West Africa in 2014 with a total of 27,741 cases and 11,284 deaths. The fever is caused by the Ebola virus, which belongs to the Filoviridae family and contains a ssRNA genome. The known subtypes of the virus are Bundibugyo ebolavirus, Reston ebolavirus, Sudan ebolavirus, $Ta\ddot{i}$ Forest ebolavirus, and Zaire ebolavirus. The Ebola outbreak was historically originated majorly from the East and Central African tropical belt. The current outbreaks in West Africa caused numerous deaths and spread fear in global society. In the absence of effective treatment strategies and any vaccine, accurate diagnosis is the most important contributing factor in the management and control of the epidemic disease. WHO (World Health Organization) has announced emergency guidance for the selection and use of Ebola in in vitro diagnostic assays. Numerous companies and research institutions have studied the various diagnosis methods and identified four WHO procurement approved as diagnosis kits: RealStar Ebolavirus Screen RT-PCR kit 1.0 (Altona), Liferiver-Ebola Virus (EBOV) Real time RT-PCR kit, Xpert Ebola Assay, and ReEBOV Antigen Rapid Test Kit. The efficiency of novel diagnostic kits such as Rapid Diagnosis Test (RDT) is currently being evaluated.

Evaluation of Commercial Complementary DNA Synthesis Kits for Detecting Human Papillomavirus (인유두종바이러스 검출을 위한 상용화된 cDNA 합성 키트의 평가)

  • Yu, Kwangmin;Park, Sunyoung;Chang, Yunhee;Hwang, Dasom;Kim, Geehyuk;Kim, Jungho;Kim, Sunghyun;Kim, Eun-Joong;Lee, Dongsup
    • Korean Journal of Clinical Laboratory Science
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    • v.51 no.3
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    • pp.309-315
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    • 2019
  • Cervical cancer is the fourth most common malignant neoplasm in women worldwide. Most cases of cervical cancer are caused by an infection by the human papillomavirus. Molecular diagnostic methods have emerged to detect the HPV for sensitivity, specificity, and objectivity. In particular, real-time PCR has been introduced to acquire a more sensitive target DNA or RNA. RNA extraction and complementary DNA synthesis are proceeded before performing real-time PCR targeting RNA. To identify an adequate and sensitive cDNA synthesis kit, this study evaluated the two commonly used kits for cDNA synthesis. The results show that the $R^2$ and efficiency (%) of the two cDNA synthesis kits were similar in the cervical cancer cell lines. On the other hand, the Takara kit compared to Invitrogen kit showed P<0.001 in the $10^2$ and $10^3$ SiHa cell count. The Takara kit compared to the Invitrogen kit showed P<0.001 in the $10^1$ and $10^2$ HeLa cell count. Furthermore, 8, 4, 2, 1, and 0.5 ml of forty exfoliated cell samples were used to compare the cDNA synthesis kits. The Takara kit compared to the Invitrogen kit showed P<0.01 in 8, 4, and 1 ml and P<0.05 in 0.5 mL. The study was performed to identify the most appropriate cDNA synthesis kit and suggests that a cDNA synthesis kit could affect the real-time PCR results.