• Title, Summary, Keyword: serological diagnosis(ELISA)

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Pathological and serological detection of bovine viral leukosis in a dairy farm in Jeonbuk province (유우농장에서 발생한 소바이러스성 백혈병의 병리학적 및 혈청학적 조사)

  • Jo Young-Suk;Jang Sae-Gun;Chu Keum-Suk;Choi Eun-Young;Chon Hee-Woong;Hong Jae-Hee;Lim Chae-Woong
    • Korean Journal of Veterinary Service
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    • v.29 no.2
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    • pp.89-96
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    • 2006
  • Bovine viral leukosis is a viral disease of cattle characterized by the development of tumors in the lymphatic tissue. A female Holstein, 3-year-old, was submitted for diagnosis at the Diagnostic laboratory, Chonbuk National University. Clinical sign of the affected animal showed emaciation, enlargement of superficial lymph node and mild diarrhea. Remarkable lesions were enlargement of many internal lymph nodes. Histopathology revealed excessive neoplastic lymphoid cells characteristic of BVL infection. Subsequently, serums from all cattle were collected and serological examination was done where a 85% seropositive rate was detected using ELISA test. ELISA method showed a comparatively 75% higher detection rate than the agar gel immunodiffusion (AGID) test (85% vs 40%). Serologically positive cattle were variably detected in all ages from under 1 year to over 6 year of age. Hematological examination consistently showed leukocytosis and a differential lymphocytosis of seropositive cattle. Detailed comparative pathological and serological data diagnosed the presence of bovine viral leukosis.

Nested-PCR and a New ELISA-Based NovaLisa Test Kit for Malaria Diagnosis in an Endemic Area of Thailand

  • Thongdee, Pimwan;Chaijaroenkul, Wanna;Kuesap, Jiraporn;Na-Bangchang, Kesara
    • The Korean Journal of Parasitology
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    • v.52 no.4
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    • pp.377-381
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    • 2014
  • Microscopy is considered as the gold standard for malaria diagnosis although its wide application is limited by the requirement of highly experienced microscopists. PCR and serological tests provide efficient diagnostic performance and have been applied for malaria diagnosis and research. The aim of this study was to investigate the diagnostic performance of nested PCR and a recently developed an ELISA-based new rapid diagnosis test (RDT), NovaLisa test kit, for diagnosis of malaria infection, using microscopic method as the gold standard. The performance of nested-PCR as a malaria diagnostic tool is excellent with respect to its high accuracy, sensitivity, specificity, and ability to discriminate Plasmodium species. The sensitivity and specificity of nested-PCR compared with the microscopic method for detection of Plasmodium falciparum, Plasmodium vivax, and P. falciparum/P. vivax mixed infection were 71.4 vs 100%, 100 vs 98.7%, and 100 vs 95.0%, respectively. The sensitivity and specificity of the ELISA-based NovaLisa test kit compared with the microscopic method for detection of Plasmodium genus were 89.0 vs 91.6%, respectively. NovaLisa test kit provided comparable diagnostic performance. Its relatively low cost, simplicity, and rapidity enables large scale field application.

Applicability of ABC-ELISA and Protein A-ELISA in serological diagnosis of cysticercosis (유구낭미충증의 혈청학적 진단을 위한 ABC-ELISA와 Protein A-ELISA의 유용성)

  • Lee, Jong-Hyun;Kong, Yoon;Ryu, Jae-Young;Cho, Seung-Yull
    • The Korean Journal of Parasitology
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    • v.31 no.1
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    • pp.49-56
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    • 1993
  • Specific antibody test in serum and cerebrosinal fluid (CSF) is still the main mode of serological diagnosis of cystiercosis. Of different techniques of artibody test, enzyme-linked immunosorbent assay (micro-ELISA) has widely been applied. This study was undertaken to observe whether diagnostic capability can be Improved by applying more sensitive techniques such as Protein A-ELISA and avidin biotin complex ELISA (ABC-ELISA). When evaluated using 115 sera of human cysticercosis, the antibody positive rates were not significantly improved in Protein A-ELISA (82.6%) and in ABC-ELISA (86.1%) than in micro-ELISA (81.7%). The specificities, evaluated in 165 sera from other diseases and normal controls, were significantly improved (88.5% by micro-ELISA, 93.3% by Protein A-ELISA and 93.8% by ABC-ELISA). Antibody levels (absorbance, abs.) in individual serum were correlated well (r : 0.83∼0.86) each other. An actual benefit of Protein A-ELISA and ABC-ELISA was that they needed smaller amount of test sample.

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Isolation and comparison of somatic and excretory-secretory antigens for serological diagnosis in Trichuris suis (혈청학적 진단을 위한 돼지 편충의 체항원, 배설/분비항원의 분리 및 비교)

  • Jee, Cha-ho;Lee, Chul-soon;Park, Seung-jun
    • Korean Journal of Veterinary Research
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    • v.39 no.1
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    • pp.159-168
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    • 1999
  • Swine whipworm(Trichuris suis) is cosmopolitan nematode which can cause serious pathology in immature stage(larva2~larva5) of infected pigs, such as anorexia, diarrhea, anemia, and death in heavy infections. In this larval stages, it is very difficult to diagnose the infection of whipworm and to differentiate from other common swine gastrointestinal disorders such as 21 day scours which are associated with TGE virus, rota virus, coccidium, and the stress of weaning. In this experiment, the isolated antigens of Trichuris spp. were carried out to examine the structure and specificity of antigens and to select the reasonable antigens which would be used in serological diagnosis by electrophoresis, Western blotting, ELISA. The results of this experiment were as follows; 1. The common fractions of each Trichuris suis antigen were identified 28,32,45, 80kDa by SDS-PAGE with silver stain and four major fractions could be detected in positive swine sera by Western blot analysis. 2. The OD(optical density) values of somatic and excretory-secretory antigens which were reacted against positive(negative) sera from pigs infected with Trichuris suis by ELISA reader were; 1) OD values($mean{\pm}SD$) of adult somatic antigen against positive(negative) sera were $0.30{\pm}0.12(0.09{\pm}0.006)$ and third-stage larva of somatic antigen were $0.28{\pm}0.038(0.10{\pm}0.005)$. And OD values of excretory-secretory antigens of adult and third-stage larva were $0.24{\pm}0.031(0.11{\pm}0.005)$ and $0.08{\pm}0.013(0.10{\pm}0.003)$, respectively. 2) OD values of adult somatic, larval somatic antigen and adult excretory-secretory antigen response to positive sera were significantly (p<0.01) associated with negative swine sera. And the Cut-off OD values(minimum positive value) were determined to be mean negative value plus 3 SD that would minimized the risk of false positives. 3. The OD values of somatic antigens of T suis and T vulpis against swine positive(negative) sera were $0.30{\pm}0.120(0.09{\pm}0.006)$ and $0.25{\pm}0.141(0.09{\pm}0.003)$. These data mean that the somatic antigens of T suis and T vulpis were able to diagnose T vulpis infection in dogs as well as T suis infection in pigs. These results suggest that somatic antigen of third-stage larva and excretory-secretory antigen of adult T suis could be used the diagnostic antigen by serological test(ELISA) in immature Trichuris spp. infection.

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Diagnosis of porcine reproductive and respiratory syndrome (PRRS) and its serological survey using the reverse transcription and polymerase chain reaction (RT-PCR) and ELISA (RT-PCR과 ELISA를 이용한 PRRS 진단 및 항체가 조사)

  • Chu Keum-Suk;Han Keu-Sam;Han Jae-Cheol;Song Hee-Jong
    • Korean Journal of Veterinary Service
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    • v.27 no.3
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    • pp.273-280
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    • 2004
  • The studies were performed for the PRRS antigen and antibody detection from breeding farms, artificial insemination(AI) center and growing farms in Jeonbuk province. 1. Specific PRRS primers were successfully amplified ORF6 617bp and ORF7 448 bp on agarose gel. 2. RT-PCR method has been establish by commercial kit and the thermal cycler program consisted of 30 cycles: $95^{\circ}C$ for 30 sec, $45^{\circ}C$ for 30 sec, and $72^{\circ}C$ for 45 sec. 3. The results of PRRS antibody test by ELISA method in AI centers were $6.6\%,\;53.3\%$ and breeding farms $65\%,\;65\%\;and\;38.7\%$, respectively. The serological positive of the antibody in gilt higher than sow. 4. The sero-positive of the PRRS antibody showed average $21\%$ in domestic farms, $56.2\%$ in breeding farms, and $29.9\%$ in AI center.

Intraerythrocytic culture and development of serological diagnostic tests of Babesia gibsoni 1. Indirect fluorescent antibody test and enzyme-linked immunosorbent assay for antibody detection of Babesia gibsoni infections in dogs (Babesia gibsoni의 적혈구내 배양법과 진단법 개발에 관한 연구 1. Babesia gibsoni 진단을 위한 간접형광항체법(IFAT)과 효소표지면역검사법(ELISA))

  • Suh, Myung-deuk;Shin, Yong-seung
    • Korean Journal of Veterinary Research
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    • v.37 no.3
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    • pp.583-593
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    • 1997
  • Indirect fluorescent antibody test(IFAT) and enzyme-linked imuunosorbent assay (IgG-ELISA) as serological diagnostic tools were conducted to evaluate the usefulness for diagnosis of canine babesiosis infected with Babesia gibsoni in domestic various dog breeds, american pit bullterrier, military shepherd, and mongrel dogs. The results obtained from this study were abstracted as follows. The nonionic detergent Triton X-100 and absorbent bio-bead $SM_2$ were useful reagents for the preparation of pure merozoite antigen of B gibsoni to be used in ELISA. The optimum reaction in ELISA was shown when the protein concentration of ELISA antigen was measured as 625ng/ml and the conjugate concentration was diluted into 1/6000 fold. The average OD value of ELISA in sera determined with negative responses in IFAT was measured as $0.255{\pm}0.051$(490nm) and the cut - off value of OD was determined as 0.399(490nm). The serum antibodies in both of IFAT and ELISA were detected on one week after artificially infected with B gibsoni and these high antibody titers, 512X in IFAT and 1024X in ELISA, were long lasted until 15 weeks after infection. The reproducibility of reaction and stability of the antigen absorbed microtitration polystyrene plate preserved in $4^{\circ}C$ refrigerator and $-20^{\circ}C$ freezer, respectively could be lasted until 135 days after storage. The positive rates in IFAT by dog breeds were shown 8.1%(60/744 heads) in mongrel dogs, 81.3%(78/96 heads) in american pit bullterrier and 15.6%(15/96 heads) in military shepherd, while the positive rate in ELISA shown 17.6%(131/744 heads) in mongrel dogs, 83.3%(80/96 heads) in american pit bullterrier and 36.5%(35/96 heads) in military shepherd, respiectively. In the total of 936 heads surveyed with IFAT and ELISA the positive rates in IFAT and ELISA were 16.4%(153/936 heads) and 26.3%(246/936 heads), respectivily. Agreement of reactions between IFAT and ELISA was shown 82.4% in 936 dog sera. The specificity and sensitivity of ELISA reaction were 83.5% and 76.5%, respectively. From the conclusion obtained in this study it was evaluated that IFAT and ELISA were useful as highly specific, sensitive and stable serelogical tools for the diagnosis of canine babesiosis in Korea.

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ELISA detection of vivax malaria with recombinant multiple stage-specific antigens and its application to survey of residents in endemic areas

  • Kim, Sera;Ahn, Hye-Jin;Kim, Tong-Soo;Nam, Ho-Woo
    • The Korean Journal of Parasitology
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    • v.41 no.4
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    • pp.203-207
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    • 2003
  • An ELISA was developed for the diagnosis of vivax malaria using multiple stage-specific recombinant antigens of Plasmodium vivax. The DNA from the whole blood of a malaria patient was used as template to amplify the coding regions for the antigenic domains of circumsporozoite protein (CSP-1), merozoite surface protein (MSP-1), apical merozoite antigen (AMA-1), serine repeat antigen (SERA), and exported antigen (EXP-1). Each amplified DNA fragment was inserted into pQE30 plasmid to induce the expression of His-tagged protein in Escherichia coli (M15 strain) by IPTG. His-tagged proteins were purified by Ni-NTA metal-affinity chromatography and used as antigens for ELISA with patient sera that were confirmed previously by blood smear examinations. When applied to patient sera, 122 (80.3%) out of 152 vivax malaria cases reacted to at least one antigen, while no reactions were observed with 128 uninfected serum samples. We applied this ELISA to the screening of 3,262 civilian residents in endemic regions near the DMZ, which resulted in 236 positively detected (7.2%) cases. This method can be applied to serological diagnosis and mass screening in endemic regions, or can be used as a safety test for transfusion blood in endemic areas.

Evaluation of Two ELISA and Two Indirect Hemagglutination Tests for Serodiagnosis of Pulmonary Hydatid Disease

  • Eris, Fatma Nur;Akisu, Ciler;Aksoy, Umit
    • The Korean Journal of Parasitology
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    • v.47 no.4
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    • pp.427-429
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    • 2009
  • To establish a definite diagnosis for pulmonary hydatid disease, combination of radiology and serology is useful. In this study, 19 preoperative sera from patients with surgically confirmed pulmonary hydatidosis, 40 sera from patients with other parasitosis and pulmonary diseases, and 20 sera from healthy donors were evaluated using 4 different serological tests, i.e., the commercial ELISA (ELISA-kit) test, the ELISA (ELISA-lab) test prepared in our laboratory, the commercial indirect hemagglutination assay kit (IHA-kit) test, and the IHA test using sensitized sheep red blood cells with tannic acid (IHA-TA). The ELISA-kit was the most sensitive (84.2%) and the most specific test (100.0%). The ELISA-kit also demonstrated the highest positive (100.0%) and negative (95.2%) predictive values. The sensitivity of the ELISA-lab test, that we prepared, was found to be 73.6%, whereas the IHA-kit test and the IHA-TA test were found to be 73.6% and 68.4%, respectively. The specificity of these tests was 96.6%, 98.3%, and 83.3%, respectively. When all 4 tests were assessed together, it was found that the sensitivity had risen to 94.7%. When the ELISA-kit was assessed with the IHA-kit and IHA-TA together, it was found that the sensitivity was 89.5% and 84.2%, respectively. Likewise, the combination of the ELISA-lab and IHA-kit or IHA-TA allowed us to achieve a sensitivity of 84.2% in cases of pulmonary echinococcosis. In conclusion, the diagnosis would be imminent if least 2 tests were applied together.

Diagnosis and seroprevalence of porcine respiratory coronavirus disease (돼지호흡기코로나바이러스 감염증의 감별진단과 항체분포 조사)

  • Kim, Eun-Gyeong;Son, Byeong-Kuk;Lee, Jong-Min;Kim, Tho-Kyoung
    • Korean Journal of Veterinary Service
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    • v.32 no.4
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    • pp.293-298
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    • 2009
  • Porcine respiratory coronavirus (PRCV) is antigenically related to transmissible gastroenteritis virus (TGEV). Differential serological diagnosis between PRCV and TGEV infection is not possible with the classical sero-neutralization test. Infection with PRCV or TGEV induces antibodies which neutralize both viruses to the same titer. However, the enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR) can differentiate between PRCV and TGEV infection. This study was carried out to investigate the prevalence of PRCV infection of swine in Gyeongnam province. A total of 391 serum samples from 37 herds in Gyeongnam were examined for antibody to PRCV using blocking ELISA. All serum samples were collected from 130- to 150-day-old pigs between August and December 2006. By ELISA, 182 out of 391 sera tested (46.5%) and 29 out of 37 sample herds (78.4%) were positive against PRCV. Our data suggested that seropositive herds for PRCV are distributed diffusely throughout Gyeongnam. The PCR methods were established to diagnose PRCV spike protein (S) gene. PCR were conducted to identify the PRCV genome against 150 pigs in PRCV antibody positive herds.

The survey of Trichinella spiralis infection in finishing pigs using the pepsin-digestion method and ELISA in Korea (조직인공소화법과 ELISA를 이용한 국내 출하돈의 선모충(Trichinella spiralis) 감염실태 조사)

  • Seo, Hunsu;Woo, Gye-Hyeong;Youn, Hee-Jeong
    • Korean Journal of Veterinary Research
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    • v.44 no.2
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    • pp.269-277
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    • 2004
  • Trichinella spiralis is one of the important zoonotic parasites with a wide variety of vertebrates hosts in nature. The purpose of this study were to analyze ESP(Excretory-Secretory Protein) antigen, to evaluate ELISA for the serological diagnosis of Trichinosis, and to survey T. spiralis infection in finishing pigs using the pepsin digestion method and ELISA in Korea. In the analysis of ESP antigen by SDS-PAGE and Western blot, 4 major bands (70, 55, 52.6, and 49 kDa) were revealed from the ESP antigen. Predilection sites of T. spiralis were the diaphragm, the tongue, masseter muscles, intercostal muscle, and hindlimb in orders in the experimentally infected rats. Sera from 581 swine were tested by ELISA with ESP antigen. The 54 (9.3%) sera were suspected as positive reactors, however, these 54 sera were determined as false positives by the use of Western blotting. This study demonstrated that the ELISA was not suitable for the examination of T. spiralis in pork. The diaphragm muscle samples of 251 finishing pigs were tested by the method of pepsin-digestion for the presence of Trichinella larvae, however, T. spiralis was not detected from the samples. We could not find out T. spiralis infection in pig in Korea pork.