scholarly journals Dynamics of Antigenemia and Coproantigens during a Human Fasciola hepatica Outbreak

1998 ◽  
Vol 36 (9) ◽  
pp. 2723-2726 ◽  
Author(s):  
Ana M. Espino ◽  
Ailén Díaz ◽  
Antonio Pérez ◽  
Carlos M. Finlay
Keyword(s):  
Fasciola Hepatica ◽  
Clinical Symptoms ◽  
Sandwich Elisa ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
La Palma ◽  
Detection Assay ◽  
Circulating Antigen ◽  
Circulating Antigens ◽  
Stool Samples

In the present study the dynamics of antigenemia and coproantigens were studied in patients withFasciola hepatica infection during an outbreak occurring in La Palma, Pinar del Rı́o, in the West Province of Cuba. Stool and serum samples were collected from 67 patients and 40 healthy subjects. Stool samples were studied by a simple gravity sedimentation technique and an ES78 sandwich enzyme-linked immunosorbent assay (ELISA) for observation of eggs and detection of parasite coproantigens, respectively. Serum samples were also studied by the ES78 sandwich ELISA and an indirect ELISA to detect circulating antigens and antibodies, respectively. At the beginning of the study, 8 of 67 patients had patent infections and 59 had prepatent infections, which was determined by the recent consumption of lettuce contaminated with metacercariae of F. hepatica, the presence of clinical symptoms, and the absence of Fasciolaeggs in their stools. Patients with prepatent infections were monitored by all techniques until patency. Circulating antigens were not detected in patients with patent infections. However, coproantigens were clearly detected in all patients with patent infections. On the other hand, 28.8% of patients with prepatent infections tested positive for circulating antigens and 81.4% tested positive for coproantigens in the first stool sample studied. Only two other coproantigen determinations were necessary to diagnose 93.2% of the patients. While circulating antigen levels diminished in all patients during the infection, coproantigen levels increased. The present study demonstrates that the ES78 sandwich ELISA is a better tool than parasitological examination for diagnosis of active early infection, since by the combination of the circulating-antigen detection assay and the coproantigen detection assay 91% of patients were able to be diagnosed at the beginning of the study. In contrast, a coprologic analysis repeated over several weeks was necessary to diagnose 100% of the patients.

scholarly journals Immunodiagnosis of cattle fascioliasis using a 27 kDa Fasciola gigantica antigen

2021 ◽  
pp. 2097-2101
Author(s):  
Mohamed J. Saadh ◽  
Samer A. Tanash ◽  
Ammar M. Almaaytah ◽  
Issam J. Sa'adeh ◽  
Saed M. Aldalaen ◽  
...  
Keyword(s):  
Western Blotting ◽  
Clinical Symptoms ◽  
Characteristic Curve ◽  
Fasciola Gigantica ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infected Cattle ◽  
Serum Samples ◽  
Healthy Cattle ◽  
Target Epitope ◽  
Circulating Antigens

Background and Aim: Diagnosis of fascioliasis depends on clinical symptoms and routine laboratory tests. Recently, antibodies and circulating antigens of Fasciola were used for detecting active infections. Therefore, this study aimed to identify Fasciola gigantica antigens in the sera of infected cattle using Western blotting and enzyme-linked immunosorbent assay (ELISA) for an accurate diagnosis of cattle infected with F. gigantica. Materials and Methods: Serum samples were obtained from 108, 23, and 19 cattle infected with Fasciola gigantica, Paramphistomum cervi, and Strongylids, respectively, including 57 non-infected cattle that were used as healthy cattle for the study. Western blotting and ELISA were then used to detect circulating Fasciola antigens at 27 kDa. Results: The target epitope was detected in an F. gigantica adult-worm antigen preparation, excretory/secretory products, and serum from cattle infected with F. gigantica. However, it was absent in sera from P. cervi, Strongylids, and healthy cattle. The purified 27 kDa F. gigantica (FPA-27) antigen was also detected in cattle serum using ELISA with high degrees of sensitivity and specificity (94% and 82%, respectively), and the area under the receiver operating characteristic curve was 0.89 with a highly significant correlation of p<0.0001. Conclusion: The FPA-27 is proposed to be a promising candidate for the serodiagnosis of fascioliasis in cattle.

An evaluation of antigen capture assays for detecting active filarial antigens

2014 ◽  
Vol 89 (3) ◽  
pp. 352-358 ◽  
Author(s):  
R. Ravishankaran ◽  
N.S. Radhika ◽  
L. Ansel Vishal ◽  
S. Meenakshisundaram ◽  
A.A. Karande ◽  
...  
Keyword(s):  
Polyclonal Antibodies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Capture Assay ◽  
Antigen Capture ◽  
Circulating Filarial Antigens ◽  
Detection Assay ◽  
Normal Individuals ◽  
Circulating Antigen ◽  
Endemic Normal

AbstractLymphatic filariasis is a parasitic disease of tropical countries. This is a disfiguring and painful disease contracted in childhood, but the symptoms become apparent only in later years. Diagnosis of filarial infection is very crucial for the management of the disease. The main objective of this study was to develop a filarial antigen-based immunological assay for the diagnosis and surveillance of the disease. Monoclonal and polyclonal antibodies were raised to the recombinant protein Brugia malayi vespid allergen homologue (VAH). Capture enzyme-linked immunosorbent assay (ELISA) was standardized utilizing various combinations of antibodies and evaluated with serum samples of endemic normal (EN, n= 110), microfilaraemic (MF, n= 65), chronic pathology (CP, n= 45) and non-endemic normal (NEN, n= 10) individuals. Of the 230 samples tested, VAH capture assay detected circulating antigen in 97.91% of bancroftian and 100% of brugian microfilaraemic individuals, and 5% of endemic normal individuals, comparable to the earlier reported SXP-1 antigen detection assay. However, the combination of VAH and SXP-1 (VS) capture ELISA was found to be more robust, detecting 100% of microfilaraemic individuals and with higher binding values. Thus an antigen capture immunoassay has been developed, which can differentiate active infection from chronic infection by detecting circulating filarial antigens in clinical groups of endemic areas.

Assessment of monoclonal antibodies to Echinococcus granulosus Antigen 5 and Antigen B for detection of human hydatid circulating antigens

Parasitology ◽  
1993 ◽  
Vol 106 (1) ◽  
pp. 75-81 ◽  
Author(s):  
D. Liu ◽  
M. D. Rickard ◽  
M. W. Lightowlers
Keyword(s):  
Monoclonal Antibodies ◽  
Echinococcus Granulosus ◽  
Binding Capacity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Antigen B ◽  
Detection Assay ◽  
Circulating Antigens ◽  
Normal Human ◽  
Antigen 5

SUMMARYFour monoclonal antibodies (MAb) to Echinococcus granulosus Antigen 5 (Ag5) and Antigen B (AgB) were assessed in an enzyme-linked immunosorbent assay (ELISA) for detection of circulating antigens (CAg) in sera of human patients with E. granulosus infection. Around 5·5–8% of 200 sera from 42 surgically proven hydatid patients contained detectable CAg by individual MAb. The combined detection rate for CAg, using four MAb, was 19% (38/200). Although hydatid CAg was detected by MAb in at least one serum sample from 21 of 42 patients, some patients remained negative in the assay regardless of the time when serum samples were taken (pre- or post-operatively), or of the continuing presence of hydatid cysts, their location or fertility. In addition, it was observed that the binding capacity of MAb for sheep hydatid cyst fluid antigen (SHCF) was somewhat reduced in the presence of normal human serum. The CAg detection assay would only be useful for assessment of hydatid infection status in patients with detectable CAg in serum samples.

scholarly journals Detection of circulating antigens in serum samples of mice experimentally infected with Trichinella spiralis by a sandwich ELISA based on IgY

2014 ◽  
Vol 51 (3) ◽  
pp. 181-189 ◽  
Author(s):  
F. Jing ◽  
J. Cui ◽  
R. Liu ◽  
L. Liu ◽  
P. Jiang ◽  
...  
Keyword(s):  
Post Infection ◽  
Trichinella Spiralis ◽  
Sandwich Elisa ◽  
Egg Yolk ◽  
Mouse Serum ◽  
Serum Samples ◽  
Egg Yolk Immunoglobulin ◽  
Muscle Larvae ◽  
Sandwich Elisa Assay ◽  
Circulating Antigens

AbstractIn the present study, a sandwich ELISA based on IgY (egg yolk immunoglobulin) was developed for detection of circulating antigens (CAg) in sere of mice experimentally infected with Trichinella spiralis. The IgY-sandwich ELISA assay involved the use of chicken antibody IgY against excretory-secretory (ES) antigens of Trichinella spiralis muscle larvae as a capture antibody and mouse polyclonal antibody IgG to ES antigens as a detecting antibody. This method was able to detect as little as 3 ng/ml of ES antigens added to normal mouse serum. A group of sixteen mice was orally inoculated with 500 T. spiralis muscle larvae per animal. The serum samples from the infected mice were taken during 1–35 days post-infection (dpi). The CAg was detectable as early as 8 dpi in the sera of infected mice. The level of CAg increased dramatically during 13–15 dpi and reached a peak at 22 dpi and remained a plateau for 3 days, then declined gradually. Another peak of CAg occurred at 31 dpi. The anti-Trichinella antibodies was first detected in 14.3 % of the infected mice at 2 weeks post-infection (wpi), and reached a peak positive rate of 100 % at 5 wpi. Moreover, the infected mice were treated with abendazole at 5 wpi and the serum CAg levels increased significantly during 2–6 days posttreatment (dpt) and then declined rapidly during 8–14 dpt. By 42 dpt, the CAg levels decreased to the undetected level, but the detection rate of antibodies was still 100 %. The IgY-sandwich ELISA appears to be a sensitive for detection of antigenemia of T. spiralis and valuable to judge the efficacy of chemotherapy in trichinellosis.

scholarly journals Development of an Enzyme-Linked Immunosorbent Assay for Immunoglobulin M Antibodies against Measles Virus

2003 ◽  
Vol 10 (3) ◽  
pp. 439-442 ◽  
Author(s):  
F. Roodbari ◽  
M. H. Roustai ◽  
A. Mostafaie ◽  
H. Soleimanjdahi ◽  
R. Sarrami Foroshani ◽  
...  
Keyword(s):  
Immunosorbent Assay ◽  
Neutralizing Antibodies ◽  
Measles Virus ◽  
Clinical Symptoms ◽  
Maculopapular Rash ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Vaccination Programs ◽  
Elisa System

ABSTRACT Measles is a highly contagious respiratory virus infection, with typical clinical symptoms including maculopapular rash, fever, cough, coryza, and conjunctivitis. Despite implementation of widespread vaccination programs throughout the world, the rates of global morbidity and mortality are still considerable. This study was performed to design a reliable indirect enzyme-linked immunosorbent assay (ELISA) to measure measles-specific immunoglobulin M (IgM). First, human IgM was purified, and then an anti-IgM antibody was produced in rabbits and purified in a multistep process. The rabbit IgG against human IgM was conjugated with peroxidase. Measles virus-infected Vero cells produced viral antigen. One hundred serum samples from infants of 9 to 18 months of age, mostly vaccinated, were evaluated for determining the presence of specific IgM antibodies against measles virus. The samples were also evaluated for neutralizing antibodies against measles virus by a microneutralization test (MNT). By comparing the results of the ELISA with those of MNT, it was demonstrated that ELISA had a sensitivity and specificity of 100 and 92%, respectively. On the other hand, when the results obtained by our ELISA system were compared with those of an imported measles virus IgM ELISA kit (EIAgen; Adaltis Italia SPa, Bologna, Italy), a high level of agreement was shown (k = 0.926).

Sandwich enzyme immunoassay of osteocalcin in serum with use of an antibody against human osteocalcin

1993 ◽  
Vol 39 (6) ◽  
pp. 942-947 ◽  
Author(s):  
D A Monaghan ◽  
M J Power ◽  
P F Fottrell
Keyword(s):  
Monoclonal Antibody ◽  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Sandwich Elisa ◽  
Normal Subjects ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Standard Curve ◽  
Microtiter Plates ◽  
Human Osteocalcin

Abstract We have developed and thoroughly validated a solid-phase sandwich enzyme-linked immunosorbent assay (ELISA) on microtiter plates for osteocalcin in human serum with use of an antibody raised against human osteocalcin. We used a monoclonal antibody against bovine osteocalcin as the capture antibody; the second antibody was a polyclonal antibody against human osteocalcin. The amount of bound second antibody was determined with use of swine anti-rabbit antibody labeled with horseradish peroxidase. We demonstrated independence of volume and determined the recovery of added standard and within- and between-assay precision. The minimal detection limit for osteocalcin was between 1.0 and 1.5 micrograms/L and the midpoint of the standard curve ranged from 14 to 17 micrograms/L. The intraassay CV was &lt; or = 8% in the range 2.7-52 micrograms/L; the interassay CV was usually &lt; or = 15% in the same range. Analytical recovery of human osteocalcin standard added to serum samples was consistently &gt; 90%. Values for osteocalcin measured in serum from 44 normal subjects were similar to those obtained with a competitive enzyme immunoassay (EIA) that used a monoclonal antibody against bovine osteocalcin. There was a good correlation between the two assays [r2 = 0.877, slope and intercept (+/- SE) = 0.88(+/- 0.051) and 0.316(+/- 0.523), respectively]. The range and mean (+/- SD) for the sandwich ELISA and the competitive EIA were 1.7-18.1 micrograms/L [8.7(+/- 4.4) micrograms/L] and 1.9-22.8 micrograms/L [9.1(+/- 4.4) micrograms/L], respectively.

scholarly journals Human herpes virus 8 antibodies in HIV-positive patients in Surabaya, Indonesia

2020 ◽  
Author(s):  
Devi Oktafiani ◽  
Ni Luh Ayu Megasari ◽  
Elsa Fitriana ◽  
Nasronudin ◽  
Maria Inge Lusida ◽  
...  
Keyword(s):  
Drug Users ◽  
Sandwich Elisa ◽  
Human Herpesvirus ◽  
Intravenous Drug ◽  
Enzyme Linked Immunosorbent Assay ◽  
Hiv Positive ◽  
Intravenous Drug Users ◽  
Serum Samples ◽  
Serological Tests ◽  
Human Herpes Virus 8

Background: Human herpesvirus 8 (HHV-8) infection is etiologically related to Kaposi’s sarcoma. Antibodies directed against HHV-8 can be detected in 80%–95% of HIV-seropositive patients with KS. HHV-8 serological tests have been done in several countries in Southeast Asia such as Malaysia, and Thailand however no serological data is available in Indonesia. This study was to examine the presence of HHV-8 antibodies in HIV-positive patients in Surabaya, Indonesia. Material and methods: Ninety-one serum samples were collected from HIV-positive patients in Surabaya, Indonesia. Human immunodeficiency virus-positive serum samples were collected from 10 homosexual men, 25 intravenous drug users (IVDUs) and 56 heterosexuals. Serums were then tested for the presence of HHV-8 antibody by using sandwich ELISA (Abbexa Ltd, Cambridge, UK). Results: The total of 91 HIV-infected were testing with antibodies to HHV-8 using enzyme-linked immunosorbent assay. Antibodies of HHV-8 were detected in 7/91 (7.7%) of the samples. According to a gender, six men (85.7%) and a women (14.3%) were positive of HHV-8 antibodies. No correlation regarding the gender and age from this study. The antibodies of HHV-8 was detected among intravenous drug users (IVDUs) men 5/7 (42.8%) and 2/7 (28.6%) from homosexual and heterosexual, respectively. Conclusion: This study found the presence of HHV-8 antibodies in 7.7% of patients in Surabaya, Indonesia. This finding was higher more than Southeast Asian countries. The patients with a positive result could suggest measures to prevent HHV-8 infection.

Detection of circulating antigen in bancroftian filariasis by sandwich ELISA using filarial serum IgG

1984 ◽  
Vol 58 (3) ◽  
pp. 259-262 ◽  
Author(s):  
M. V. R. Reddy ◽  
Ashok Malhotra ◽  
B. C. Harinath
Keyword(s):  
Immunosorbent Assay ◽  
Sandwich Elisa ◽  
Negative Reaction ◽  
Serum Immunoglobulin ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bancroftian Filariasis ◽  
Positive Correlation ◽  
Serum Igg ◽  
Filarial Antigen ◽  
Circulating Antigen

ABSTRACTThe utility of the IgG fraction of human filarial serum immunoglobulin in detecting circulating antigen by sandwich enzyme linked immunosorbent assay (ELISA) was studied. 27 of 33 sera from persons with microfilaracmia, 19 of 30 sera from clinical cases of filariasis, 4 of 30 sera from normal persons from a region endemic for filariasis showed the presence of circulating filarial antigen. All the 20 normal sera from the area where filariasis was not endemic gave negative reaction for filarial antigen. Those sera from persons with microfilaracmia that showed the presence of circulating antigen also showed an apparent positive correlation between the microfilarial density and the antigen titre.

scholarly journals Immunodetection of Fasciola gigantica Circulating Antigen in Sera of Infected Individuals for Laboratory Diagnosis of Human Fascioliasis

2013 ◽  
Vol 20 (10) ◽  
pp. 1569-1577 ◽  
Author(s):  
Abdelfattah M. Attallah ◽  
Faisal A. Bughdadi ◽  
Atef M. El-Shazly ◽  
Hisham Ismail
Keyword(s):  
Laboratory Diagnosis ◽  
Fasciola Gigantica ◽  
Antigen Detection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Target Antigen ◽  
Immunoperoxidase Staining ◽  
Serum Samples ◽  
Content Type ◽  
Human Fascioliasis ◽  
Circulating Antigen

ABSTRACTCurrently, the laboratory diagnosis of human fascioliasis is based on the parasitological examination of parasite eggs in stool specimens and serological detection of specific antibodies in serum samples, which are often unreliable diagnostic approaches. Ideally, a sensitive and specific diagnostic test forFasciolainfection should be based on the detection of circulatingFasciolaantigen, which implies active infection. Here, a 27-kDa-molecular-mass antigen was identified in aFasciola giganticaadult worm antigen preparation, excretory-secretory products, and sera fromF. gigantica-infected individuals, and it was not detected in antigenic extracts of other parasites and sera from noninfected individuals. The target antigen was isolated and partially characterized as a protein. Immunoperoxidase staining located the target epitope within teguments and guts ofF. giganticaadult worms. The performance characteristics of a newly developed enzyme-linked immunosorbent assay (ELISA) based onF. giganticacirculating antigen detection in serum (FgCA-27 ELISA) were investigated using sera of 120 parasitologically diagnosedF. gigantica-infected individuals and 80 noninfected individuals. The area under the receiving operating characteristic (ROC) curve (AUC) for ELISA was significantly high (AUC = 0.961,P< 0.0001) for discriminatingFasciola-infected and noninfected individuals. The developed assay showed high degrees of sensitivity, specificity, and efficiency (>93%), and a significant correlation (r= 0.715,P< 0.0001) between antigen level and parasite egg count was shown. In conclusion, a 27-kDaFasciolaantigen was identified in sera ofF. gigantica-infected individuals. A highly sensitive and specificFasciolaantigen detection assay, FgCA-27 ELISA, was developed for laboratory diagnosis of human fascioliasis.

scholarly journals Evaluation and Application of an Indirect ELISA for the Detection of Antibodies to Bovine Respiratory Syncytial Virus in Milk, Bulk Milk, and Serum

1995 ◽  
Vol 7 (2) ◽  
pp. 177-182 ◽  
Author(s):  
M. Elvander ◽  
S. Edwards ◽  
IS. Näslund ◽  
N. Linde
Keyword(s):  
Respiratory Syncytial Virus ◽  
Clinical Symptoms ◽  
Bovine Respiratory Syncytial Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
National Veterinary Institute ◽  
Bulk Milk ◽  
History Of ◽  
The Difference ◽  
Syncytial Virus

An indirect enzyme-linked immunosorbent assay (ELISA) was developed at the National Veterinary Institute (NVI), Uppsala, to detect antibodies to bovine respiratory syncytial virus (BRSV) in serum and milk. For the evaluation of the NVI ELISA, field sera collected from cattle in England and Sweden were tested in parallel with an ELISA in use at the Central Veterinary Laboratory (CVL), Weybridge. The tests showed 96% agreement. The sensitivity and specificity of the NVI ELISA relative to the CVL ELISA were 94% and 100%, respectively. There was evidence that the difference in sensitivity between the 2 tests was due to the detection of both IgG and IgM class antibodies by the CVL ELISA, whereas the NVI ELISA was designed specifically to detect IgGl. Milk and serum samples from individual cows were tested by the NVI ELISA for presence of antibodies to BRSV. There was a good correlation between the ability to detect antibodies in serum and the ability to detect them in milk, although the antibody titer was generally lower in milk than in serum. Bulk milk samples were collected from farms with severe clinical symptoms of respiratory distress and from farms with no history of respiratory disease. There was a clear distinction between antibody levels in diseased and healthy herds. The NVI ELISA is a rapid and reliable test for detecting antibodies to BRSV in milk, bulk milk, and serum samples.

Sign in / Sign up

Export Citation Format

Share Document