scholarly journals Studies on the specific immunodiagnosis of cystic echinococcosis in camels using enzyme-linked immunosorbent assay

2019 ◽  
Vol 22 (3) ◽  
pp. 305-313 ◽  
Author(s):  
O. Kandil ◽  
N. Hassan ◽  
D. Sedky ◽  
E. Beshir Ata
Keyword(s):  
Cystic Echinococcosis ◽  
Haemonchus Contortus ◽  
Indirect Elisa ◽  
Cross Reactivity ◽  
Economic Losses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hydatid Cysts ◽  
Germinal Layer ◽  
Lung Cysts ◽  
Fasciola Spp

Cystic echinococcosis (CE) is of increasing public health and socio-economic concern because of the large morbidity rates and produced high economic losses in the livestock industry. The objective of the current research was to study the reliability of indirect ELISA in detecting CE, based on two dif-ferent types of crude antigens of camel origin; protoscolex and germinal layer antigens from hydatid cyst. Blood samples were collected from 284 (125 slaughtered and 159 live camels). Out of 125 slaughtered camels examined visually, 55 (44%) were found to have hydatid cysts. Of them, 52/125 (41.6%) and 3/125 (2.4%) harboured hydatid cysts in lungs and livers respectively. Fertile lung cysts were 32.8%; 26.9% were sterile, while 40.3% of lung and liver cysts were calcified. The sensitivity of ELISA was 83% and 46.5% when protoscolex and germinal layer antigens were used, respectively. The respective specificity of antigens of protoscolex and germinal layer was 70.3% and 41.7%. The protoscolex antigen showed higher accuracy (73.6%) compared to the germinal layer antigen (52.8%). The cross reactivity of these antigens were evaluated with antigens and hyperimmune sera of CE and Fasciola spp. and Haemonchus contortus using ELISA. The results showed also weak immunogenic potency of each antigen with Fasciola spp. hyperimmune sera at dilution 1:50 while hyperimmune sera of Haemonchus contortus did not bind any antigen.

scholarly journals Combined Use of Indirect ELISA and Western Blotting with Recombinant Hepatocellular Carcinoma-Associated Antigen 59 Is a Potential Immunodiagnostic Tool for the Detection of Prepatent Haemonchus contortus Infection in Goat

Animals ◽  
2019 ◽  
Vol 9 (8) ◽  
pp. 548 ◽  
Author(s):  
Muhammad Ali-ul-Husnain Naqvi ◽  
Sana Zahra Naqvi ◽  
Muhammad Ali Memon ◽  
Kalibixiati Aimulajiang ◽  
Muhammad Haseeb ◽  
...  
Keyword(s):  
Western Blot ◽  
Western Blotting ◽  
Haemonchus Contortus ◽  
Indirect Elisa ◽  
Fasciola Hepatica ◽  
False Negative ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Combined Use

Haemonchus contortus is recognized as one of the important health problems in small ruminants, leading to reduced production and economic loss for farmers worldwide. Prepatent diagnosis of H. contortus infection is crucial to improve control strategies as this helminth may remove up to one-fifth of total erythrocytes and may cause anemia, edema, diarrhea, and ultimately death in young animals. In this study, one of the excretory and secretory products, rHc-HCA59, was purified and used as antigen to detect specific antibodies in H. contortus infected goats during prepatent stage of infection using indirect enzyme linked immunosorbent assay (ELISA) as screening test. All goats (n = 38) were housed indoor, experimentally infected with 8000 infective larvae (L3) of H. contortus, and serum samples were collected prior to infection and at 14th day of infection. Immunoblotting was performed to confirm the results of indirect ELISA, evaluate the cross reactivity against rHc-HCA59 in sera of most common co-infecting parasites and rectify the false negative samples. Furthermore, three different batches of rHc-HCA59 were produced to evaluate the repeatability of ELISA. No eggs were detected in feces of all goats collected at 7th and 14th day of infection but, H. contortus eggs were detected at 21 days post infection in the feces. Indirect ELISA performed in this study showed 87% sensitivity and 100% specificity. The western blot analysis confirmed immunoreactivity in serum samples which scored positive in indirect ELISA and recognized the samples as negative which had OD450 lower than negative cut-off value in indirect ELISA. Furthermore, all false negative sera (n = 5) that had OD450 value between positive and negative cut-off value in rHc-HCA59 based ELISA were clearly positive in western blot. Moreover, no cross-reactivity was detected in ELISA and western blotting against rHc-HCA59 in positive sera of Toxoplasma gondii, Fasciola hepatica, and Trichinella spiralis. The results of this study concluded that combined use of indirect ELISA and western blotting with rHc-HCA59 is a potential immunodiagnostic tool for the detection of H. contortus infection during prepatent period in goats.

Production and Characterization of Antibodies Against Neosaxitoxin

1992 ◽  
Vol 75 (2) ◽  
pp. 341-345 ◽  
Author(s):  
Fun S Chu ◽  
Xuan Huang ◽  
Sherwood Hall
Keyword(s):  
Bovine Serum Albumin ◽  
Serum Albumin ◽  
Bovine Serum ◽  
Indirect Elisa ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
High Antibody ◽  
Antibody Titers ◽  
Antibody Elisa

Abstract Antibody against neosaxitoxin (neo-STX) was obtained from rabbits after immunization with neo- STX conjugated to either keyhole limpet hemocyanln (KLH) or bovine serum albumin (BSA). An indirect enzyme-linked Immunosorbent assay (ELISA), In which either neo-STX-BSA or neo-STXKLH was coated to the mlcroplate, was used to monitor the antibody titer. Although high antibody titers were obtained from rabbits after immunization with both immunogens, only antibody obtained from rabbits immunized with neo-STX-KLH was useful for Immunoassay. Competitive indirect ELISA revealed that the antibodies obtained from rabbits Immunized with neo-STX-KLH are specific for neo-STX but also have good cross-reactivity with STX. The concentrations causing 50% inhibition binding of neo-STX-BSA to the anti-neo-KLH by neo-STX, STX, and decarbamoyl-STX (DC-STX) were 0.9,8.0, and 53.1 ng/mL, respectively. Saxitoxin conjugated to polylyslne (STX-PLL) was also used as the coating reagent In the indirect ELISA. The concentrations causing 50% inhibition binding of antl-neo-STX-KLH to STX-PLL coated on the mlcrotiter plate by neo-STX, STX, and DC-STX were 1.2,4.1, and 36.1 ng/mL, respectively. With this newly developed antibody, ELISA could be a very effective method for monitoring seafood for both neo-STX and STX.

scholarly journals Optimization and Validation of Indirect ELISA Using Truncated TssB Protein for the Serodiagnosis of Glanders amongst Equines

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
Harisankar Singha ◽  
Praveen Malik ◽  
Sachin K. Goyal ◽  
Sandip K. Khurana ◽  
Chiranjay Mukhopadhyay ◽  
...  
Keyword(s):  
Indirect Elisa ◽  
Expression System ◽  
Cross Reactivity ◽  
Amino Acid Sequences ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Diagnostic Efficacy ◽  
Diagnostic Potential ◽  
Prokaryotic Expression System ◽  
Control Serum

Objective. To express truncated TssB protein ofBurkholderia malleiand to evaluate its diagnostic efficacy for serological detection of glanders among equines.Materials and Methods. In an attempt to develop recombinant protein based enzyme-linked immunosorbent assay (ELISA), N-terminal 200 amino acid sequences ofB. malleiTssB protein—a type 6 secretory effector protein—were expressed in prokaryotic expression system. Diagnostic potential of recombinant TssB protein was evaluated in indirect ELISA using a panel of glanders positive (n=49), negative (n=30), and field serum samples (n=1811). Cross-reactivity of the assay was assessed with equine disease control serum and human melioidosis positive serum.Results. In comparison to CFT, diagnostic sensitivity and specificity of ELISA were 99.7% and 100%, respectively.Conclusions. The indirect ELISA method using the truncated TssB offered safer and more rapid and efficient means of serodiagnosis of glanders in equines. These data highlight the use of TssB as potential diagnostic antigen for serological diagnosis of glanders.

scholarly journals CT Scanning in Identification of Sheep Cystic Echinococcosis

2017 ◽  
Vol 2017 ◽  
pp. 1-6
Author(s):  
Rui Mao ◽  
Hongzhi Qi ◽  
Lei Pei ◽  
Jie Hao ◽  
Jian Dong ◽  
...  
Keyword(s):  
Ct Scan ◽  
Cystic Echinococcosis ◽  
Ct Scanning ◽  
Lung Infection ◽  
Liver Cyst ◽  
Hydatid Cysts ◽  
Liver Cysts ◽  
Lung Cysts ◽  
Suitable Tool ◽  
Cystic Hydatid

Objective. We aim to determine the efficiency of CT in identification of cystic echinococcosis in sheep. Methods. Fifty-three sheep with liver cysts confirmed by ultrasonography were subject to CT scan to evaluate the number, size, and type of the cysts in liver and lung, confirmed using necropsy. The correlation of numbers between liver cysts and lung cysts was calculated using Pearson analysis. Results. Necropsy indicated a 98% consensus on size, location, number, and activity compared with CT scan. The viable cysts were 53.1% and 50.6% in the liver and lung, respectively. Among the cysts in liver, 35.5%, 9.5%, 5.7%, 10.2%, and 39.1% were Types CE1, CE2, CE3, CE4, and CE5, respectively. The cysts in the lungs, 17.4%, 26.9%, 12.1%, 11.6%, and 32.1%, were Types CE1, CE2, CE3, CE4, and CE5, respectively. A significant correlation was noticed between the number of cysts in liver and those in lung (R=0.770, P<0.001). Conclusions. CT scan is a suitable tool in determining the size and type of cystic hydatid cysts in both liver and lung of sheep. A significant correlation was noticed between the numbers in liver and lung, indicating that lung infection was likely due to the expansion of liver cyst burden pressure.

scholarly journals Prevalence of tick-transmitted pathogens in cattle reveals that Theileria parva, Babesia bigemina and Anaplasma marginale are endemic in Burundi

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Lionel Nyabongo ◽  
Esther G. Kanduma ◽  
Richard P. Bishop ◽  
Eunice Machuka ◽  
Alice Njeri ◽  
...  
Keyword(s):  
Spatial Distribution ◽  
Indirect Elisa ◽  
Anaplasma Marginale ◽  
Sub Saharan Africa ◽  
Economic Losses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Theileria Parva ◽  
Babesia Bigemina ◽  
Adult Cattle ◽  
Significant Difference

Abstract Background Tick-borne diseases (TBDs) constitute a major constraint for livestock development in sub-Saharan Africa, with East Coast fever (ECF) being the most devastating TBD of cattle. However, in Burundi, detailed information is lacking on the current prevalence of TBDs and on the associated economic losses from mortality and morbidity in cattle as well as the costs associated with TBD control and treatment. The aim of this study was, therefore, to assess the prevalence and spatial distribution of tick-borne pathogens (TBPs) in cattle across the major agro-ecological zones (AEZs) in Burundi. Methods In a cross-sectional study conducted in ten communes spanning the five main AEZs in Burundi, blood samples were taken from 828 cattle from 305 farms between October and December 2017. Evidence of Theileria parva infection was assessed by antibody level, measured using a polymorphic immunodominant molecule (PIM) antigen-based enzyme-linked immunosorbent assay (ELISA) and by a T. parva-specific p104 gene-based nested PCR. Antibodies against Theileria mutans infection were detected using the 32-kDa antigen-based indirect ELISA, while the 200-kDa antigen and the major surface protein 5 (MSP5)-based indirect ELISA were used to detect antibodies against Babesia bigemina and Anaplasma marginale, respectively. Results The prevalence of T. parva across the ten communes sampled ranged from 77.5 to 93.1% and from 67.8 to 90.0% based on the ELISA and PCR analysis, respectively. A statistically significant difference in infection was observed between calves and adult cattle; however, T. parva infection levels were not significantly associated with sex and breed. The seroprevalence indicating exposure to T. mutans, B. bigemina and A. marginale ranged from 30 to 92.1%, 33.7 to 90% and 50 to 96.2%, respectively. Mixed infections of TBPs were detected in 82.91% of cattle sampled, with 11 different combinations of pathogen species detected . Conclusions The findings indicate that T. parva, A. marginale and B. bigemina infections are endemic in Burundi. Knowledge of the spatial distribution of TBPs will facilitate the design of effective targeted strategies to control these diseases. There is a need for further investigations of the distribution of tick vectors and the population structure of TBPs in order to identify the key epidemiological factors contributing to TBD outbreaks in Burundi.

scholarly journals Recombinant cold shock domain containing protein is a potential antigen to detect specific antibody during early and late infections of Haemonchus contortus in goat

2020 ◽  
Author(s):  
Muhammad Ali-ul-Husnain Naqvi ◽  
KaLiBiXiaTi Aimulajiang ◽  
Muhammad Ali Memon ◽  
Muhammad Waqqas Hasan ◽  
Sana Zahra Naqvi ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Cold Shock ◽  
Fasciola Hepatica ◽  
Trichinella Spiralis ◽  
Early Stage ◽  
Cross Reactivity ◽  
Antibody Detection ◽  
Economic Losses ◽  
Diagnostic Potential ◽  
Specificity And Sensitivity

Abstract Background : Haemonchus contortus ( H. contortus ) is one of the most important parasites that cause huge economic losses to small ruminant industry worldwide. Effective prognosis and treatment depend upon the early diagnosis of H. contortus infection. To date, no widely-approved methods for the identification of prepatent H. contortus infection are available to identify prepatent H. contortus infection. The aim of this study was to evaluate the diagnostic potential of recombinant cold shock H. contortus protein (rHc-CS) during early and late infections of H. contortus in goat. Results : Purified rHc-CS exhibited a clear band, with a molecular weight about 38 kDa. H. contortus eggs were not detected by fecal egg count technique from feces collected at 0 to 14 days post infection (D.P.I). However, eggs were detected at 21, 28 and 35 D.P.I. Hence, results of immunoblotting assay showed specific anti rHc-CS antibody detection in all goat sera collected at early stage (14 D.P.I) and late stage (21-103 D.P.I) of H. contortus infection. Furthermore, no cross reactivity was observed against Trichinella spiralis , Fasciola hepatica and Toxoplasma gondii or uninfected goats. Among several evaluated rHc-CS indirect-ELISA format variables, favorable antigen coating concentration was found 0.28μg/well at 37℃ 1h and overnight at 4°C. Moreover, optimum dilution ratio of serum and rabbit anti-goat IgG was recorded as 1:100 and 1:4000, respectively. The best blocking buffer was 5% Bovine Serum Albumin (BSA) while the best time for blocking, serum incubation and TMB reaction were recorded as 60, 120 and 10 min, respectively. The cut-off value for positive and negative interpretation was determined as 0.352 (OD 450 ). The diagnostic specificity and sensitivity of the rHc-CS, both were recorded as 100%. Conclusion : These results validated that rHc-CS is a potential immunodiagnostic antigen to detect the specific antibodies during early and late H. contortus infections in goat.

scholarly journals Seroprevalence of ovine brucellosis in the microregion of Teresina, Piauí, Brazil

2020 ◽  
Vol 87 ◽  
Author(s):  
Letícia Soares de Araújo Teixeira ◽  
Joilson Ferreira Batista ◽  
Pedro Henrique Fonseca Silva ◽  
Maria Luiza Lima Cordeiro ◽  
Raissa Costa Amorim ◽  
...  
Keyword(s):  
Reproductive System ◽  
Indirect Elisa ◽  
Jugular Vein ◽  
Domestic Animals ◽  
Economic Losses ◽  
Enzyme Linked Immunosorbent Assay ◽  
Agar Gel ◽  
Serological Tests ◽  
Blood Samples ◽  
The Face

ABSTRACT: Among the diseases that affect the reproductive system of domestic animals, brucellosis in the sheep species is important because it generates significant economic losses to sheep rearing. Thus, it is a threat to the growth and productivity of sheep herds. In the face of this problem, the objective of the present research was to identify the prevalence of ovine brucellosis in herds in municipalities of the Teresina, Piauí, Brazil microregion by using the agar gel immunodiffusion assay (AGID) and indirect enzyme-linked immunosorbent assay (ELISA) serological tests. Fourteen municipalities were included in the research. Blood samples were collected from 521 pubescent animals by puncturing the jugular vein. After collection, the samples were submitted to the serological techniques, AGID and indirect ELISA, to detect anti B. ovis antibody. Of the 521 samples submitted to the AGIDtest, 23 (4.41%) were sera reagent and 498 (95.58%) negative. The indirect ELISA tests, 24 (4.61%) suspect samples and 497 (95.39%) negative samples were obtained, and there were no reagent animals in this test, only suspect. The seroprevalence of ovine brucellosis in the Teresina, Piauí microregion was 4.41%. Thus, it is possible to identify sheep with reagent serology to infection by B. ovis, that is present in municipalities in the state of Piauí, Brazil. Furthermore, AGIDwas shown to be more sensitive in detecting animals that had had contact with the etiological agent of the disease.

scholarly journals Epidemiological, Morphometric, and Molecular Investigation of Cystic Echinococcosis in Camel and Cattle From Upper Egypt: Current Status and Zoonotic Implications

2021 ◽  
Vol 8 ◽  
Author(s):  
Ahmed Gareh ◽  
Amira A. Saleh ◽  
Samar M. Moustafa ◽  
Amin Tahoun ◽  
Roua S. Baty ◽  
...  
Keyword(s):  
Hydatid Cyst ◽  
Molecular Identification ◽  
Cystic Echinococcosis ◽  
Current Status ◽  
Economic Losses ◽  
Organ Distribution ◽  
Hydatid Cysts ◽  
Transmission Cycle ◽  
Parasitic Zoonoses ◽  
Female Cattle

Cystic echinococcosis has been considered one of the major parasitic zoonoses which is associated with severe economic losses. The present study was undertaken to investigate the occurrence, organ distribution, cyst fertility, and viability of cystic echinococcosis in slaughtered camels and cattle from various abattoirs in Assiut Governorate, Egypt. The work also involved morphological, morphometric, and molecular identification of the parasite. The occurrence of hydatid cysts was investigated in total number of 100 lungs of camels and 574 liver and lungs of cattle admitted to three slaughterhouses at Assiut Governorate, Egypt. Moreover, several individual variable factors, including organ involvement, age, sex, and hydatid cyst characteristics, were studied to identify their possible association with the occurrence of the disease. Genomic DNA was extracted from the hydatid cysts, followed by molecular identification of the parasite through amplification of ribosomal DNA internal transcribed spacer (ITS) regions. Hydatid cysts were found in 6 camels (6%) out of 100 inspected camels, while 5 hydatid cysts (0.87%) were detected in a total number of 574 cattle examined. The parasite was detected exclusively in lungs of camels, while lungs were the main organ infected by the parasite in cattle and one hydatid cyst was found in the liver (0.17%). In camel, 66.7, 16.65, and 16.65%of detected cysts were fertile, sterile, and calcified, respectively, while in cattle, these percentages were 60, 20, and 20%, respectively. None of the studied variable factors were significantly associated with the occurrence of the disease in camels, with the exception that all cysts were found in the lung. Conversely, we found a significant association (P &lt; 0.05) between the age and sex of the slaughtered cattle and the occurrence of hydatid cysts. In this respect, the rate of infection was higher in female cattle and those cattle more than 5 years (P &lt; 0.05). The morphological, morphometric, and molecular studies confirmed the presence of the parasite. Taken together, our results concluded that camels and cattle play a potential role in maintaining the transmission cycle of this zoonotic parasite.

scholarly journals Development and Potential Application of Ras Domain Containing Protein from Haemonchus contortus for Diagnosis of Goat Infection

Animals ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 138 ◽  
Author(s):  
Kalibixiati Aimulajiang ◽  
Man Cao ◽  
Shuyi Liao ◽  
Muhammad Ali-ul-Husnain Naqvi ◽  
Xiaowei Tian ◽  
...  
Keyword(s):  
Haemonchus Contortus ◽  
Indirect Elisa ◽  
Fasciola Hepatica ◽  
Trichinella Spiralis ◽  
Clinical Signs ◽  
Small Ruminants ◽  
Gastrointestinal Nematode ◽  
Enzyme Linked Immunosorbent Assay ◽  
Fecal Examination ◽  
Diagnostic Potential

Haemonchus contortus is an important gastrointestinal nematode of small ruminants that causes significant mortality in goats worldwide. Diagnosis of this infection mainly depends on the evaluation of clinical signs and fecal examination. However, limitations often occur in early or mild infections. For this purpose, serological diagnosis seems to be more accurate and reliable. Ras domain-containing protein (Ras) is one of H. contortus’s excretory and secretory products (ESPs) that can be isolated from different larval stages of the nematode. In this study, the recombinant H. contortus Ras domain-containing protein (rHcRas) was expressed and purified and its diagnostic potential was evaluated. Reactions between rHcRas and goat sera were tested using Western blotting (WB). The results showed that rHcRas could be recognized by sera as early as 14 days post infection (DPI), and antibodies against rHcRas in infected goats could be maintained for over 89 days. No reaction was found between rHcRas and antibodies against Trichinella spiralis, Fasciola hepatica, or Toxoplasma gondii. An indirect enzyme-linked immunosorbent assay (ELISA) was produced based on rHcRas. The optimal coating antigen (157 ng of rHcRas/well) and serum dilutions (1:50) were determined via checkerboard titration. Indirect ELISA based on rHcRas showed 87.5% sensitivity and 90.6% specificity. The cut-off values for this experiment were determined to be 0.324 (positive) and 0.273 (negative), respectively, and the variation coefficient (CV) was less than 15%. The results of the indirect ELISA in-field examination showed that 17.6% (9/51) of the goats were infected with H. contortus, higher than the fecal examination results (15.7%, 8/51). When compared the results of the indirect ELISA and necropsy testing, 98.0% (50/51) consistency was found. These results indicated that rHcRas was a potential antigen for the diagnosis of H. contortus infection in goats.

scholarly journals Limitations of the BP26 Protein-Based Indirect Enzyme-Linked Immunosorbent Assay for Diagnosis of Brucellosis

2013 ◽  
Vol 20 (9) ◽  
pp. 1410-1417 ◽  
Author(s):  
Ting Xin ◽  
Hongjun Yang ◽  
Nan Wang ◽  
Fang Wang ◽  
Peng Zhao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Beef Cattle ◽  
Yersinia Enterocolitica ◽  
Immunosorbent Assay ◽  
Indirect Elisa ◽  
Cross Reactivity ◽  
Infected Sheep ◽  
Enzyme Linked Immunosorbent Assay ◽  
Content Type ◽  
Experimental Animals

ABSTRACTBrucellosis is a serious zoonosis that occurs worldwide, and its diagnosis is typically based on the detection of antibodies againstBrucellalipopolysaccharide (LPS). However, the specificity of the LPS-based test is compromised by cross-reactivity withEscherichia coliO157:H7 andYersinia enterocoliticaO:9. Also, diagnosis based on the LPS test cannot differentiate between vaccinated and infected individuals. The detection of the 26-kDa cytosoluble protein (BP26) antibody is considered an alternative that circumvents these drawbacks because it is exclusively expressed by infectiousBrucella. A BP26-based enzyme-linked immunosorbent assay (ELISA) has been tried for the diagnosis ofBrucella-infected animals and humans, but a few results showed that BP26 couldn't react with allBrucella-positive sera. In order to explore whether different animals could produce antibodies against BP26 after being infected with variousBrucellaspecies, we infected sheep, goats, and beef cattle with common virulent referenceBrucellaspecies. All sera were collected from the experimental animals and tested using both LPS-based ELISAs and BP26-based ELISAs. The results showed that allBrucella-infected individuals could produce high levels of antibodies against LPS, but onlyB. melitensis16M- andB. melitensisM28-infected sheep andB. melitensis16M- andB. abortus2308-infected goats could produce antibodies against BP26. Therefore, we concluded that the BP26-based indirect ELISA (i-ELISA) showed bothBrucellaspecies and host specificity, which obviously limits its reliability as a substitute for the traditional LPS-based ELISA for the detection of brucellosis.

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