Expression of sheep pathogen Babesia sp. Xinjiang rhoptry-associated protein 1 and evaluation of its diagnostic potential by enzyme-linked immunosorbent assay

Parasitology ◽  
2016 ◽  
Vol 143 (14) ◽  
pp. 1990-1999 ◽  
Author(s):  
QINGLI NIU ◽  
ZHIJIE LIU ◽  
JIFEI YANG ◽  
PEIFA YU ◽  
YUPING PAN ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Post Infection ◽  
Small Ruminants ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Diagnostic Potential ◽  
Positive Rate ◽  
Ct Antigen ◽  
High Level ◽  
Potential Use

SUMMARYOvine babesiosis is one of the most important tick-borne haemoparasitic diseases of small ruminants. The ovine parasite Babesia sp. Xinjiang is widespread in China. In this study, recombinant full-length XJrRAP-1aα2 (rhoptry-associated protein 1aα2) and C-terminal XJrRAP-1aα2 CT of Babesia sp. Xinjiang were expressed and used to evaluate their diagnostic potential for Babesia sp. Xinjiang infections by indirect enzyme-linked immunosorbent assay (ELISA). Purified XJrRAP-1aα2 was tested for reactivity with sera from animals experimentally infected with Babesia sp. Xinjiang and other haemoparasites using Western blotting and ELISA. The results showed no cross-reactivities between XJrRAP-1aα2 CT and sera from animals infected by other pathogens. High level of antibodies against RAP-1a usually lasted 10 weeks post-infection (wpi). A total of 3690 serum samples from small ruminants in 23 provinces located in 59 different regions of China were tested by ELISA. The results indicated that the average positive rate was 30·43%, and the infections were found in all of the investigated provinces. This is the first report on the expression and potential use of a recombinant XJrRAP-1aα2 CT antigen for the development of serological assays for the diagnosis of ovine babesiosis, caused by Babesia sp. Xinjiang.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay for the Detection of Antibody to Epizootic Hemorrhagic Disease of Deer Virus

2000 ◽  
Vol 12 (2) ◽  
pp. 142-145 ◽  
Author(s):  
James O. Mecham ◽  
Michael M. Jochim
Keyword(s):  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hemorrhagic Disease ◽  
Structural Protein ◽  
Serum Samples ◽  
Epizootic Hemorrhagic Disease ◽  
Diagnostic Potential ◽  
The Us ◽  
Serotype 1 ◽  
Infected Cells

An enzyme-linked immunosorbent assay has been developed to detect antibodies to epizootic hemorrhagic disease of deer virus (EHDV). The assay incorporates a monoclonal antibody to EHDV serotype 2 (EHDV-2) that demonstrates specificity for the viral structural protein, VP7. The assay was evaluated with sequential sera collected from cattle experimentally infected with EHDV serotype 1 (EHDV-1) and EHDV-2, as well as the four serotypes of bluetongue virus (BTV), BTV-10, BTV-11, BTV-13, and BTV-17, that currently circulate in the US. A competitive and a blocking format as well as the use of antigen produced from both EHDV-1-and EHDV-2-infected cells were evaluated. The assay was able to detect specific antibody as early as 7 days after infection and could differentiate animals experimentally infected with EHDV from those experimentally infected with BTV. The diagnostic potential of this assay was demonstrated with field-collected serum samples from cattle, deer, and buffalo.

scholarly journals Point Seroprevalence Survey of Ehrlichia ruminantium Infection in Small Ruminants in The Gambia

2005 ◽  
Vol 12 (4) ◽  
pp. 508-512 ◽  
Author(s):  
Bonto Faburay ◽  
Susanne Munstermann ◽  
Dirk Geysen ◽  
Lesley Bell-Sakyi ◽  
Ansumana Ceesay ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Small Ruminants ◽  
The Gambia ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
The West ◽  
Ehrlichia Ruminantium ◽  
The North ◽  
North Bank ◽  
Western Division

ABSTRACT Using the MAP1-B enzyme-linked immunosorbent assay, we tested 1,318 serum samples collected from sheep and goats at 28 sites in the five divisions of The Gambia to determine the Ehrlichia ruminantium seroprevalence rates and to assess the risk for heartwater. About half (51.6%) of 639 sheep were positive, with seroprevalence rates per site varying between 6.9% and 100%. The highest seroprevalence was detected in the western part of the country (88.1% in the Western Division and 62.1% in the Lower River Division). Sheep in the two easterly divisions (Central River and Upper River divisions) showed the lowest seroprevalence of 29.3% and 32.4%, respectively, while those in the North Bank Division showed an intermediate prevalence of 40.6%. In goats, less than one-third (30.3%) of 679 animals tested were positive. The highest seroprevalence was detected in goats in the North Bank Division (59%) and Western Division (44.1%). Goats in the Lower River Division showed an intermediate level of 21.9%, whereas the lowest rates were found in the eastern part of the country (4.8% in the Central River Division and 2.3% in the Upper River Division). At nearly all sites, seroprevalence rates were higher in sheep than in goats. The results show a gradient of increasing heartwater risk for susceptible small ruminants from the east to the west of The Gambia. These findings need to be taken into consideration when future livestock-upgrading programs are implemented.

scholarly journals Rapid Competitive Enzyme-Linked Immunosorbent Assay for Detection of Antibodies to Peste des Petits Ruminants Virus

2005 ◽  
Vol 12 (4) ◽  
pp. 542-547 ◽  
Author(s):  
Kang-Seuk Choi ◽  
Jin-Ju Nah ◽  
Young-Joon Ko ◽  
Shien-Young Kang ◽  
Nam-In Jo
Keyword(s):  
Immunosorbent Assay ◽  
Small Ruminants ◽  
Viral Disease ◽  
Turnaround Time ◽  
Enzyme Linked Immunosorbent Assay ◽  
Peste Des Petits Ruminants ◽  
Rinderpest Virus ◽  
Serum Samples ◽  
Specificity And Sensitivity ◽  
Relative Specificity

ABSTRACT Peste des petits ruminants (PPR) is a contagious viral disease of small ruminants that is of economic importance in Africa, the Middle East, and Asia. We developed a rapid competitive enzyme-linked immunosorbent assay (rapid c-ELISA) for the diagnosis and surveillance of PPR. This assay detects PPR virus (PPRV) antibodies in serum samples by quantifying the amount of monoclonal antibody (MAb) P-3H12 after 30 min of incubation of a serum-MAb conjugate mixture on plates coated with a PPRV recombinant nucleocapsid protein (rPPRV-N). We tested 249 PPRV-positive serum samples and 733 PPRV-negative serum samples from field ruminants. The threshold of percent inhibition (PI) was determined to be <50 on the basis of the mean PI plus 3 standard deviations for sera from PPRV-negative ruminants. The relative specificity and sensitivity of the rapid c-ELISA were 98.5% (722 of 733 serum samples) and 93.4% (234 of 249 serum samples), respectively. The rapid c-ELISA sensitively detected PPRV antibodies in hyperimmune sera (virus neutralization test [VNT] titer, >512), even at dilutions ≥512 in normal goat serum, and as early as 6 to 13 days postinfection from 12 goats, each of which was infected with one of the four PPRV lineages. Hyperimmune sera from animals experimentally vaccinated with rinderpest virus gave positive results by the rapid c-ELISA when the rinderpest virus VNT titers were >512, although the rapid c-ELISA titers were very low (2 to 16). However, the rapid c-ELISA was negative when the rinderpest virus VNT titer was ≤128. The rapid c-ELISA developed in the present work provides a short turnaround time and could be a useful tool for the diagnosis of PPR and screening for PPRV in the field.

Evaluating the Babesia bovis infection of cattle in China with enzyme-linked immunosorbent assay (ELISA)

2015 ◽  
Vol 60 (4) ◽  
Author(s):  
Congshan Yang ◽  
Junlong Liu ◽  
Anyan Li ◽  
Youquan Li ◽  
Aihong Liu ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Control Strategies ◽  
Babesia Bovis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bovine Babesiosis ◽  
Serum Samples ◽  
The World ◽  
Positive Rate ◽  
Cattle Serum ◽  
The Mean

AbstractBabesia bovis is an important pathogen of bovine babesiosis and causes serious constraints on the health and productivity of domestic cattle in the tropical and subtropical areas of the world. Aiming to clarify the prevalence of B. bovis in China, a total of 2,364 cattle serum samples were randomly collected from 45 different areas of 17 provinces in China. Antibodies against B. bovis were tested by enzyme-linked immunosorbent assay (ELISA) using a recombinant C-terminal antigen of B. bovis rhoptry-associated protein-1 (RAP-1) to evaluate the prevalence of B. bovis. The results showed that the parasite was present in all of the investigated 17 provinces. The positive rate was from 6.40 to 47.27%, and the mean rate was 24.92%. These survey data will provide important information for designing control strategies for bovine babesiosis in China.

scholarly journals Characterization of Toxoplasma gondii SAG2 Expressed in Insect Cells by Recombinant Baculovirus and Evaluation of Its Diagnostic Potential in an Enzyme-Linked Immunosorbent Assay

2002 ◽  
Vol 9 (6) ◽  
pp. 1343-1347 ◽  
Author(s):  
Xiaohong Huang ◽  
Xuenan Xuan ◽  
Hiroshi Suzuki ◽  
Chihiro Sugimoto ◽  
Hideyuki Nagasawa ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Immunosorbent Assay ◽  
Insect Cells ◽  
Fluorescent Antibody ◽  
Recombinant Baculovirus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Conformational Structure ◽  
Serum Samples ◽  
Weak Reaction ◽  
Diagnostic Potential

ABSTRACT A baculovirus carrying the SAG2 gene of Toxoplasma gondii was constructed, and recombinant SAG2 protein (S-rSAG2) was expressed in insect cells. S-rSAG2 was recognized by sera from cats and pigs infected with T. gondii. Mice immunized with S-rSAG2 produced high titers of specific immunoglobulin G2a (IgG2a) and IgG1 antibodies. In an indirect fluorescent antibody test, all mouse antisera against S-rSAG2 reacted strongly to the natural parasites, but those against rSAG2 expressed in Escherichia coli (E-rSAG2) only showed very weak reaction, although no markedly difference was found in the reaction to denatured antigen, T. gondii lysate, in Western blot analysis. The results suggest that S-rSAG2 is better than E-rSAG2 in both antigenicity and immunogenicity. Enzyme-linked immunosorbent assay (ELISA) with S-rSAG2 could differentiate clearly between sera from 30 specific-pathogen-free cats and 4 experimentally infected cats. Serum samples from domestic cats in Japan were tested by the ELISA and compared with a latex agglutination test (LAT) and ELISA with E-rSAG2. Of 187 samples, all 35 LAT-positive sera had strong reactions to S-rSAG2 and E-rSAG2. Of the 152 LAT-negative sera, 18 were positive in the ELISA with S-rSAG2, whereas only 2 were positive in the ELISA with E-rSAG2. Although there were significant correlations among the three methods, the ELISA with S-rSAG2 was more sensitive than the others, which could be attributed to the fact that S-rSAG2 shares some common conformational structure with the native antigen. The results suggest that S-rSAG2 would be a useful reagent for the detection of T. gondii infection in cats.

scholarly journals Diagnosis of Oesophagostomum columbianum infection in goat by indirect enzyme linked immunosorbent assay

2010 ◽  
Vol 47 (2) ◽  
pp. 83-87 ◽  
Author(s):  
R. Jas ◽  
J. Ghosh ◽  
K. Das
Keyword(s):  
Immunosorbent Assay ◽  
Post Infection ◽  
Economic Losses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Oesophagostomum Columbianum ◽  
Infected Goat ◽  
Third Stage ◽  
Binding Antibody ◽  
Sensitivity Specificity

AbstractOesophagostomosis is one of the most prevalent nematodosis with considerable economic losses in small ruminant livestock. Prepatent stage of Oesophagostomum columbianum is primarily involved in the major pathogenic effect of the infection and conventional methods are unable to detect this stage. An indirect enzyme linked immunosorbent assay (ELISA) was therefore standardized and evaluated for the purpose to improve diagnosis and thereafter minimize the economic losses due to this infection in goat. The eggs obtained from O. columbianum adult female worms, collected from caecum and colon of slaughtered goats, were cultured to obtain the third stage larvae (L3). Crude somatic antigen (CSAg) was prepared from triturated worms. Experimental goats (n = 12) were orally infected each with 600 L3 /kg body weight. Serum samples of the infected goats were collected at 2-day-interval from day-3 till day-33 post infection. The indirect ELISA was standardized using the CSAg for coating the wells, infected goat sera as binding antibody, anti-goat IgG-HRP conjugate and ortho-phenylenediamine (OPD) with peroxide as substrate. The sensitivity, specificity and accuracy of ELISA were determined by testing 96 serum samples of goats whose parasitological status after slaughter was determined by examination of the abomasums and the intestines. O. columbianum antibodies were detected in sera of infected goats on day 18 – 27 post-infection with 96.00, 73.91 and 85.42 per cent sensitivity, specificity and accuracy, respectively.

scholarly journals A Highly Sensitive and Subspecies-Specific Surface Antigen Enzyme- Linked Immunosorbent Assay for Diagnosis of Johne's Disease

2006 ◽  
Vol 13 (8) ◽  
pp. 837-844 ◽  
Author(s):  
Shigetoshi Eda ◽  
John P. Bannantine ◽  
W. R. Waters ◽  
Yasuyuki Mori ◽  
Robert H. Whitlock ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Surface Antigens ◽  
Johne's Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Diagnostic Specificity ◽  
Serum Samples ◽  
Low Level ◽  
Specificity And Sensitivity ◽  
Highly Sensitive ◽  
High Level

ABSTRACT Johne's disease (JD), or paratuberculosis, caused by Mycobacterium avium subsp. paratuberculosis, is one of the most widespread and economically important diseases of livestock and wild ruminants worldwide. Control of JD could be accomplished by diagnosis and good animal husbandry, but this is currently not feasible because commercially available diagnostic tests have low sensitivity levels and are incapable of diagnosing prepatent infections. In this study, a highly sensitive and subspecies-specific enzyme-linked immunosorbent assay was developed for the diagnosis of JD by using antigens extracted from the surface of M. avium subsp. paratuberculosis. Nine different chemicals and various intervals of agitation by vortex were evaluated for their ability to extract the surface antigens. Various quantities of surface antigens per well in a 96-well microtiter plate were also tested. The greatest differences in distinguishing between JD-positive and JD-negative serum samples by ethanol vortex enzyme-linked immunosorbent assay (EVELISA) were obtained with surface antigens dislodged from 50 μg/well of bacilli treated with 80% ethanol followed by a 30-second interval of agitation by vortex. The diagnostic specificity and sensitivity of the EVELISA were 97.4% and 100%, respectively. EVELISA plates that had been vacuum-sealed and then tested 7 weeks later (the longest interval tested) had diagnostic specificity and sensitivity rates of 96.9 and 100%, respectively. In a comparative study involving serum samples from 64 fecal culture-positive cattle, the EVELISA identified 96.6% of the low-level fecal shedders and 100% of the midlevel and high-level shedders, whereas the Biocor ELISA detected 13.7% of the low-level shedders, 25% of the mid-level shedders, and 96.2% of the high-level shedders. Thus, the EVELISA was substantially superior to the Biocor ELISA, especially in detecting low-level and midlevel shedders. The EVELISA may form the basis for a highly sensitive and subspecies-specific test for the diagnosis of JD.

scholarly journals The First Seroprevalence Investigation of Peste Des Petits Ruminants Virus among Sahel Goat in Yobe State, Nigeria

2020 ◽  
pp. 33-38
Author(s):  
Babagana Alhaji Bukar ◽  
Abdul-Dahiru El-Yuguda ◽  
Lawal Said
Keyword(s):  
Local Government ◽  
Immunosorbent Assay ◽  
Small Ruminants ◽  
Viral Disease ◽  
Enzyme Linked Immunosorbent Assay ◽  
Seroprevalence Rate ◽  
Peste Des Petits Ruminants ◽  
Serum Samples ◽  
Significant Difference ◽  
Sahel Goat

Peste des petits ruminants is among the most common viral disease conditions of small ruminants, whose status has not yet been reported in Yobe State, Nigeria. Thus, this study was aimed at determining the seroprevalence of this disease among Sahel goats in Yobe State, Nigeria, using competitive enzyme-linked immunosorbent assay (c-ELISA). Out of 460 serum samples collected, 255/460 (55.4%) were positive for PPR antibodies. Seroprevalence rates of 56.1%, 55.4% and 54.6% were recorded in Bursari, Bade and Nangere Local Government Areas (LGAs) respectively. There was no statistically significant difference (p>0.05) observed in the PPRV seroprevalence rates among the three LGAs. Sahel goats older than 18 months had a significantly higher (p<0.0001) Sero-prevalence of 65.2% compared to the 35.3% observed among younger ones (<18 months). The sex-wise distribution of the Peste des petits ruminants virus (PPRV) seroprevalence rate showed that female Sahel goats had 60.0% and the males had 44.6%. The detection of the PPRV among Sahel goats from all the LGAs sampled suggests that PPRV is endemic in the study area. It is therefore recommended that PPR vaccination be instituted in the study areas.

scholarly journals Comparative Performance of a Novel Herpes Simplex Virus Type 2-Specific Enzyme-Linked Immunosorbent Assay Using a Targeted Chain Oligopeptide, Peptide 55

2009 ◽  
Vol 16 (6) ◽  
pp. 931-934 ◽  
Author(s):  
A. M. Al-Sulaiman ◽  
P. J. Vallely ◽  
P. E. Klapper
Keyword(s):  
Herpes Simplex Virus ◽  
Herpes Simplex ◽  
Immunosorbent Assay ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igg Antibody ◽  
Comparative Performance ◽  
Simplex Virus ◽  
High Level

ABSTRACT Herpes simplex virus (HSV) glycoprotein G (gG2) has been used as the basis of many serological assays for the detection of HSV type 2 (HSV-2)-specific antibodies. In the present study, an enzyme-linked immunosorbent assay (ELISA), the Pathozyme Viro HSV-2 immunoglobulin G (IgG) ELISA (Omega Diagnostics, Alva, United Kingdom), based on an immunodominant epitope of gG2 presented in a branched-chain format (peptide 55), was compared with two commercially available gG2-specific assays, the Bioelisa HSV-2 IgG assay (Biokit, S.A., Barcelona, Spain) and the HerpesSelect HSV-2 IgG assay (Focus Diagnostics, Cypress, CA). A panel of 218 well-characterized serum samples was tested. Thirty-one samples were determined to be HSV-2 IgG antibody positive and 164 samples were determined to be negative with all three kits. The levels of concordance between the tests were 95.9% between the Omega and HerpeSelect assays, 90.8% between the Omega and Bioelisa assays, and 94.5% between the HerpeSelect and Bioelisa assays. Twenty-three samples gave discordant results. Western blot results showed that of these, the results for 77% were correctly identified by the Omega assay, the results for 68% were correctly identified by the HerpeSelect assay, and the results for 13.6% were correctly identified by the Bioelisa assay. Although there was a high level of agreement between the results obtained by the three assays and no false-positive results were detected by any of the three kits, confirmation of the results for samples with discordant results by Western blotting suggested that the peptide 55-based Omega assay is the most sensitive and specific assay among the assays evaluated.

scholarly journals Serological Investigation of Peste Des Petits Ruminants in East Shewa and Arsi Zones, Oromia Region, Ethiopia

2017 ◽  
Vol 2017 ◽  
pp. 1-5 ◽  
Author(s):  
Getachew Gari ◽  
Biressaw Serda ◽  
Dejene Negesa ◽  
Fethu Lemma ◽  
Hagos Asgedom
Keyword(s):  
Significant Variation ◽  
Small Ruminants ◽  
Control Measures ◽  
Economic Losses ◽  
Enzyme Linked Immunosorbent Assay ◽  
High Seroprevalence ◽  
Serum Samples ◽  
Male And Female ◽  
Significant Difference ◽  
Important Disease

Peste des petits ruminant (PPR) is an economically important disease of small ruminants with a rapidly expanding geographical distribution. There are fragmented reports to the occurrence and distribution of the disease in Ethiopia. A total of 700 serum samples were collected from goats and sheep to detect the presence of antibody against PPR virus using Competitive Enzyme-Linked Immunosorbent Assay (C-ELISA). An overall PPR seropositivity was reported to be 48.43% in the area. There is no statistically significant difference in the seroprevalence of the disease between sheep and goats (50.85% and 46.68%), respectively. However, there was statistically significant variation (P<0.05) in the seroprevalence of the disease in young (33.9%) and adult (55.8%) age categories. The seroprevalence in male and female was 42.07% and 50.09%, respectively, where the variation was statistically not significant (P>0.05). High seroprevalence of Peste des petites ruminants in the study area indicated the virus circulation and endemicity of the disease. The disease causes substantial economic losses by affecting the livelihood of the farmers. Therefore, control measures should be put in place to minimize the loss associated with the disease.

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