• Title/Summary/Keyword: ELISA kit

Search Result 338, Processing Time 0.029 seconds

Developing Polyclonal Antibody-based Indirect-ELISA to Detect Anthracnose Inocula Prior to Soybean Sprout Rot

  • Park, Soo-Bong;Lim, Young-Ji;Lee, Jung-Han;Han, Ki-Soo;Lee, Sun-Cheol;Shim, Chang-Ki;Kang, Jin-Ho;Bae, Dong-Won;Kim, Dong-Kil;Kim, Hee-Kyu
    • The Plant Pathology Journal
    • /
    • v.20 no.4
    • /
    • pp.252-257
    • /
    • 2004
  • We developed a polyclonal antibody based-ELISA system to monitor inocula accurately and rapidly before onset of anthracnose on soybean sprouts. Titer of mouse antisera against conidia of Colletotrichum gloeosporioides, determined by indirect ELISA, was high enough to be detectable up to ${\times}$25,600 dilutions. Both PAb1 and PAb2 had the highest level of reactivity to Colletotrichum gloeosporioides. Absorbance readings exceeded 0.15. Sensitivity of PAb to C. gloeosporioides was precise enough to detect spore concentration as low as 500 conidia/well by indirect ELISA. Both antibodies are very sensitive and highly specific to the target pathogen Colletotrichum gloeosporioides, apparently discriminating other unrelated pathogen, or epiphytes. This kit fulfills the requirements far detecting inocula before infection and onset of anthracnose. Our ELISA system should also be feasible to detect C. acutatum (Mungbean sprouts rot) and G. cingulata (C. gleosporioides), (apple, pepper). It was remarkable that absorbance value was not reduced even after 4 consecutive washings (Fig.4), suggesting that antigenic determinants are on the surface of conidia. Antigenic determinant was characterized by heating and enzyme treatment: Both PAb1 and PAb2 bind to protein epitope that does not contain residue of amino acid, arginine, and Iysine, even though more work needs to be done.

Dirofilaria immitis infection in wild raccoon dogs, Nyctereutes procyonoides (야생너구리 (Nyctereutes procyonoides)의 개 심장사상충 (Dirofilaria immitis) 감염)

  • Song, Kun-Ho;Lee, Il-Berm;Kim, Duck-Hwan
    • Korean Journal of Veterinary Research
    • /
    • v.42 no.4
    • /
    • pp.545-550
    • /
    • 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.

Seroprevalence of Neospora caninum in dairy cattle of northern Gyeonggi province in Korea (경기도 북부지역 젖소의 Neospora caninum 항체 양성률 조사)

  • Jung, Kwang;Shim, Hang-Sub
    • Korean Journal of Veterinary Service
    • /
    • v.36 no.1
    • /
    • pp.53-56
    • /
    • 2013
  • This study was carried out to investigate the seroprevalence of Neospora caninum infection in dairy cattle of northern Gyeonggi province in Korea. A total of 716 dairy cattle from 14 dairy farms were tested for the presence of antibodies against N. caninum using a commercial ELISA test kit. Herd and individual seroprevalences of N. caninum were 92.8% (13/14) and 23.1% (166/716), respectively. The seroprevalence of dairy cattle according to age was the highest at 49~72 months (27.4%), but the difference was not statistically significant (P>0.05). No significant relationship between N. caninum seropositivity with the herd size of the dairy farms was found (P>0.05). The results indicate that N. caninum infection spreads widely in dairy farms of northern Gyeonggi province in Korea.

High seroprevalence of Ornithobacterium rhinotracheale in layer chickens in Gyeonggi province, South Korea (경기도 산란계에서 Ornithobacterium rhinotracheale의 높은 항체 양성률)

  • Jung, Kwang
    • Korean Journal of Veterinary Service
    • /
    • v.43 no.4
    • /
    • pp.257-259
    • /
    • 2020
  • Ornithobacterium rhinotracheale (ORT) causes pneumonia, airsacculitis, and pleuritis in chickens and other avian species. Little is known about the seroprevalence of ORT in layer chickens in Gyeonggi province, South Korea. The purpose of this study was to determine the seroprevalence of ORT in layer chickens in Gyeonggi province, South Korea from May to September 2019. A total of 460 chickens in 28 flocks were tested for antibodies to ORT by using commercial enzyme-linked immunosorbent assay (ELISA) kit. The seroprevalence of ORT antibodies in the flocks was 100% (28/28) and the overall seroprevalence in individual chickens was 98.91% (455/460). This survey indicated the high seroprevalence of ORT in layer chickens in Gyeonggi province, South Korea. Therefore, measures should be executed to control ORT in layer chickens in this province.

The Application of ParalluxTM System for Multi-Detection of (Fluoro)quinolone Class Antibiotics Residues in Raw Bovine Milk

  • Park, Hong-Je;Kim, Gyung-Dong;Han, Kyu-Ho;Lee, Chi-Ho
    • Food Science of Animal Resources
    • /
    • v.33 no.2
    • /
    • pp.198-204
    • /
    • 2013
  • This study aimed to apply the Parallux system to detect (fluoro)quinone antibiotics residues in raw bovine milk. The immunogen enabled the generation of a specific antiserum with a titer of 1/40,000. The $Parallax^{TM}$ kit using the antibody displayed $IC_{50}$ value of 10 to 150 ppb for (fluoro)quinolone antibiotics. $Parallax^{TM}$ kit was also sensitive for the detection of incurred (fluoro)quinolone at Korean Maximum Residual Levels in raw bovine milk as the result of dose response test. Cross reactivities of the antibody with the common (fluoro)quinolones were determined to be norfloxacin, 100%; enrofloxacin, 100%; ciprofloxacin, 100%; danofloxacin, 100%; nalidixic acid, 40%. Lower detection limit (LOD) values of the $Parallax^{TM}$ kit in raw bovine milk were determined to be norfloxacin, 4 ppb; enrofloxacin, 5 ppb; danofloxacin, 5 ppb; ciprofloxacin, 5 ppb and nalidixic acid, 10 ppb. The $Parallax^{TM}$ kit was run 8 times with five different concentrations of norfloxacin to determine the coefficient of variation (CV, %) of intra-assay, which was between 2.7% and 11.8%. To confirm the precision among kit batches for the inter-assay, five different batch kits were tested with 2 different concentration of norfloxacin. The CVs of the inter assay were 4.2% at 50 ppb, and 7.2% at 10 ppb norfloxacin, respectively.

Synthetic Peptide-Based Enzyme-Linked Immunosorbent Assay for Human $\alpha$-Fetoprotein

  • Yoon, Mi-Chung;Lee, Hyun-Hee
    • Biomedical Science Letters
    • /
    • v.7 no.3
    • /
    • pp.103-110
    • /
    • 2001
  • $\alpha$-Fetoprotein(AFP) is a good marker for the detection of several diseases such as hepatocellular carcinoma, gonadal germ cell tumor, gastric tumor, and Down's syndrome. In this study, we developed ELISA, using synthetic peptides corresponding to the epitopes of AFP. Five kinds of peptides were synthesized from AFP to produce antibodies in rats that recognize AFP in human plasma as well as amniotic fluid and do not cross-react with serum albumin. All five kinds of antibodies showed good reactivities with their peptide-keyhole limpet hemocyanin conjugates. Anti-synthetic peptide 1 (R-N-E-Y-G-I-A-S-I-L, 4-13) antibody, in particular, reacted well with AEP as well as synthetic peptide 1-KLH but not with human serum albumin. The binding affinity(Kd) was 2.7$\times$10$^{-9}$M for peptide 1 and 6.8$\times$10$^{-8}$M for AEP. The range for measurement of AFP was 10~1,000 ng/ml. The within-assay and between-assay coefficients of variance(CV) were 4.83% and 10.97%, respectively. In a sample of 31 sera and 33 amniotic fluids, there was a good correlation between AFP values determined in this assay and those in a commercial kit. These results indicate that the antibodies against synthetic peptides corresponding to the epitopes of AFP are highly specific to APP and synthetic peptide-based ELISA would be useful for the measurement of human AFP.

  • PDF

Seroepidemiological study of Toxoplasma gondii infection in the rural area Okcheon-gun, Korea

  • Lee, Young-Ha;Hoh, Hyung-Jun;Hwang, Ok-Sun;Lee, Sang-Keol;Shin, Dae-Whan
    • Parasites, Hosts and Diseases
    • /
    • v.38 no.4
    • /
    • pp.251-256
    • /
    • 2000
  • There have been some reports about the prevalence of anti-Toxoplasma gondii antibody among Koreans, and most of all data were taken from patients visiting hospitals. However, the epidemiological data of the community-based study in Korea are rare. This study was performed to evaluate the seroprevalence of toxoplasmosis among the inhabitants of the rural area Okcheon-gun, Korea. A total of 1,109 serum samples (499 males, 610 females) were examined for the IgG antibodies by ELISA. To set up the cut-off point for ELISA, we used a commercial latex agglutination (LA) kit. The sensitivity and specificity of ELISA against LA test were 89.5%, and 98.6% respectively. Among 1,109 sera, 6.9% showed seropositivity by ELISA. The positive rates of males and females were 6.0% and 7.2%, respectively. However, there were no significant differences between sexes. Comparing the age groups, the highest seropositive rate showed in the seventies or higher, and their rates had a tendency to increase with age (0.05 < p < 0.3). These results revealed that the seroprevalence of toxoplasmosis in rural inhabitants is similar to previous reports in Korea; however we need further investigation to clarify the prevalence of toxoplasmosis in the general population.

  • PDF

Evaluation of Recombinant SAG1, SAG2, and SAG3 Antigens for Serodiagnosis of Toxoplasmosis

  • Khanaliha, Khadijeh;Motazedian, Mohammad Hossein;Kazemi, Bahram;Shahriari, Bahador;Bandehpour, Mojgan;Sharifniya, Zarin
    • Parasites, Hosts and Diseases
    • /
    • v.52 no.2
    • /
    • pp.137-142
    • /
    • 2014
  • Serologic tests are widely accepted for diagnosing Toxoplasma gondii but purification and standardization of antigen needs to be improved. Recently, surface tachyzoite and bradyzoite antigens have become more attractive for this purpose. In this study, diagnostic usefulness of 3 recombinant antigens (SAG1, SAG2, and SAG3) were evaluated, and their efficacy was compared with the available commercial ELISA. The recombinant plasmids were transformed to JM109 strain of Escherichia coli, and the recombinants were expressed and purified. Recombinant SAG1, SAG2, and SAG3 antigens were evaluated using different groups of sera in an ELISA system, and the results were compared to those of a commercial IgG and IgM ELISA kit. The sensitivity and specificity of recombinant surface antigens for detection of anti-Toxoplasma IgG in comparison with commercially available ELISA were as follows: SAG1 (93.6% and 92.9%), SAG2 (100.0% and 89.4%), and SAG3 (95.4% and 91.2%), respectively. A high degree of agreement (96.9%) was observed between recombinant SAG2 and commercial ELISA in terms of detecting IgG anti-Toxoplasma antibodies. P22 had the best performance in detecting anti-Toxoplasma IgM in comparison with the other 2 recombinant antigens. Recombinant SAG1, SAG2, and SAG3 could all be used for diagnosis of IgG-specific antibodies against T. gondii.

Infection status of bovine viral diarrhea virus in Korean native cattle in Jeonbuk State, Korea (전북지역 한우의 소 바이러스성 설사 바이러스 감염 현황 조사)

  • Woo Ri Jung;Mi Seon Kang;Keum-Suk Chu
    • Korean Journal of Veterinary Service
    • /
    • v.47 no.2
    • /
    • pp.73-79
    • /
    • 2024
  • Bovine viral diarrhea virus (BVDV) is distributed in cattle worldwide and causes significant economic losses to the livestock industry. Identification and remove of BVDV persistently infected (PI) cattle is very important to control BVDV infection in cattle herd. The objective of this study is to investigate the infection status of BVDV infection in Korean native cattle (Bos taurus coreanae) farms located in Jeonbuk State. From 2021 to 2022, a total of 1,497 samples were collected from 17 cattle farms and tested for BVDV antigen using a commercial ELISA kit. By the first ELISA testing, 24 cattle from six farms were positive for BVDV antigen, showing the farm-level or cattle-level prevalence of 35.3% or 1.6%, respectively. By the second ELISA testing which carried out with the first ELISA-positive samples after three-weeks, 12 cattle (0.8%) from five farms (29.4%) were positive for BVDV antigen, indicating these cattle were PI cattle. Genotypes of BVDV were determined with 12 BVDV-positive samples using a previously described RT-PCR assay and the results showed that 3 (25.0%) and 9 (75.0%) were confirmed to be type 1 and type 2, respectively. These results will be helpful to establish the effective control strategy for BVDV in cattle farms in Jeonbuk State.