scholarly journals Western blot assay of anti-Echinococcus granulosus antibody positive serum samples by indirect haemagglutination method

2019 ◽  
Vol 76 (2) ◽  
pp. 195-202 ◽  
Author(s):  
Ayşe Semra Güreser ◽  
Gamze Gizem Duman ◽  
Fakhriddin Sarzhanov ◽  
Djursun Karasartova ◽  
Funda Dogruman Al ◽  
...  
Keyword(s):  
Western Blot ◽  
Echinococcus Granulosus ◽  
Serum Samples ◽  
Western Blot Assay ◽  
Indirect Haemagglutination ◽  
Positive Serum

scholarly journals Interlaboratory Optimization and Evaluation of a Serological Assay for Diagnosis of Human Baylisascariasis

2013 ◽  
Vol 20 (11) ◽  
pp. 1758-1763 ◽  
Author(s):  
Lisa N. Rascoe ◽  
Cynthia Santamaria ◽  
Sukwan Handali ◽  
Sriveny Dangoudoubiyam ◽  
Kevin R. Kazacos ◽  
...  
Keyword(s):  
Western Blot ◽  
Cross Reactivity ◽  
Western Blot Assay ◽  
Content Type ◽  
National Reference ◽  
Baylisascaris Procyonis ◽  
Reference Centre ◽  
Assay Performance ◽  
Positive Serum ◽  
Control And Prevention

ABSTRACTA Western blot assay using a recombinant protein, recombinantBaylisascaris procyonisRAG1 protein (rBpRAG1), was developed for the diagnosis of human baylisascariasis concurrently by the Centers for Disease Control and Prevention (CDC) in Atlanta, Georgia, and the National Reference Centre for Parasitology (NRCP) in Montreal, Canada. Assay performance was assessed by testing 275 specimens at the CDC and 405 specimens at the NRCP. Twenty specimens from 16 cases of baylisascariasis were evaluated. Eighteen were positive, with the assay correctly identifying 14 of 16 patients. The rBpRAG1 Western blot assay showed no cross-reactivity withToxocara-positive serum and had an overall sensitivity of 88% and a specificity of 98%.

Evaluation of reactivity to Echinococcus spp. among rural inhabitants in Poland

2015 ◽  
Vol 60 (3) ◽  
Author(s):  
Ewa Cisak ◽  
Jacek Sroka ◽  
Angelina Wójcik-Fatla ◽  
Violetta Zając ◽  
Jacek Dutkiewicz
Keyword(s):  
Western Blot ◽  
Echinococcus Granulosus ◽  
Cross Reactivity ◽  
Farm Animals ◽  
Control Group ◽  
Laboratory Diagnostics ◽  
Group 14 ◽  
Serum Samples ◽  
The City ◽  
Echinococcus Spp

AbstractA group of 172 rural inhabitants from eastern Poland (68 males and 104 females, mean age 49.0 ± 12.0 years) was examined for the presence of antibodies against Echinococcus granulosus and Echinococcus multilocularis. A population of 38 healthy urban dwellers from the city of Lublin (17 males and 21 females, mean age 36.2 ± 9.6 years) were examined as a control group. Sera of 22 rural inhabitants (12.8%) reacted positively to Echinococcus granulosus hydatid fluid antigen in the screening test. A cross-reactivity was observed with two serum samples that tested positive in ELISA for E. granulosus. Three serum samples were tested positive for E. multilocularis using the Em2plus ELISA assay and also positive for Western blot. None of the members of control group showed the presence of a seropositive reaction to Echinococcus spp. The reactivity to Echinococcus spp. among rural inhabitants decreased with age and this correlation was statistically significant (R = -0.197151, p = 0.009535). The percentage of positive findings was the highest (50.0%) in the youngest age group (14-20). No significant correlations were found between responses to interview questions (possession of domestic and farm animals, contact with wild animals, eating unwashed berries, drinking unboiled water) and the presence of seropositive reactions to Echinococcus spp. The presented results seem to indicate that echinococcosis is still a current problem in Poland that should not be neglected and, moreover, indicates the need for improvement in the routine laboratory diagnostics of Echinococcus spp. by standardizing the ELISA and Western blot tests.

Epidemiological and clinical features of HEV infection: a survey in the district of Foggia (Apulia, Southern Italy)

2013 ◽  
Vol 142 (2) ◽  
pp. 287-294 ◽  
Author(s):  
G. SCOTTO ◽  
D. MARTINELLI ◽  
M. CENTRA ◽  
M. QUERQUES ◽  
F. VITTORIO ◽  
...  
Keyword(s):  
General Population ◽  
Western Blot ◽  
Blood Donors ◽  
Southern Italy ◽  
Hepatitis E ◽  
Hiv Positive ◽  
Italian Population ◽  
Serum Samples ◽  
Western Blot Assay ◽  
Low Prevalence

SUMMARYIn this study we assessed the seroprevalence of hepatitis E virus (HEV) infection in both the Italian population and immigrants from developing countries in Foggia (Apulia, Southern Italy). The seroprevalence of HEV was determined in 1217 subjects [412 (34%) immigrants and 805 Italian subjects (blood donors, general population, HIV-positive, haemodialysis patients)]. Serum samples were tested for anti-HEV and confirmed by Western blot assay; in positive patients HEV RNA and genotype were also determined. There were 8·8% of patients that were positive to anti-HEV, confirmed by Western blot. The prevalence in immigrants was 19·7%, and in Italians 3·9% (blood donors 1·3%, general population 2·7%, HIV-positive patients 2·0%, haemodialysis patients 9·6%). Anti-HEV IgM was found in 38/107 (35·5%) of the anti-HEV-positive serum samples (34 immigrants, four Italians). This study indicates a higher circulation of HEV in immigrants and Italian haemodialysis patients, whereas a low prevalence of HEV antibodies was seen in the remaining Italian population.

scholarly journals Enzyme Immunoassay Detection of Antigen-Specific Immunoglobulin G Antibodies in Longitudinal Serum Samples from Patients with Cryptosporidiosis

2001 ◽  
Vol 8 (2) ◽  
pp. 415-423 ◽  
Author(s):  
Jeffrey W. Priest ◽  
Anna Li ◽  
Mohamad Khan ◽  
Michael J. Arrowood ◽  
Patrick J. Lammie ◽  
...  
Keyword(s):  
Western Blot ◽  
Immunoglobulin G ◽  
Protozoan Parasite ◽  
Epidemiologic Studies ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Igg Antibody ◽  
Large Format ◽  
Western Blot Assay ◽  
Wide Range

ABSTRACT Cryptosporidium parvum is a protozoan parasite that causes diarrheal illness in a wide range of mammalian hosts, including humans. Characteristic serum immunoglobulin G (IgG) antibody responses to antigens in the 27- and 17-kDa size ranges have been shown to develop after infection, and several enzyme-linked immunosorbent assay (ELISA) and Western blot assay formats have been used to measure these IgG levels in human serum. Using a collection of serial samples from laboratory-confirmed cryptosporidiosis patients, we compared the results obtained by using two new ELISAs with those obtained with two different Western blot assays. When assayed with the large-format Western blot, 97% of the 67 patients had a demonstrable antibody response on at least one occasion. The Cp23 ELISA correctly identified 93% of the samples that had a 27-kDa response by Western blot and 100% of the negative samples. The Triton antigen ELISA detected 77% of the samples that had a 17-kDa response by Western blot and 88% of the negative samples. The sensitivity of the Triton antigen assay was higher for samples collected between 16 and 92 days after the onset of symptoms (96%). The minigel-format Western blot did not compare favorably with the large-format blot for the detection of antibodies to the 27-kDa antigen (71% sensitivity). A half-life of about 12 weeks was estimated for antibodies to both the 27- and 17-kDa antigens. We believe the Cp23 and Triton antigen ELISAs will be useful in epidemiologic studies of the prevalence ofCryptosporidium infection in the population.

scholarly journals Development of a Western Blot Assay for Detection of Bovine Immunodeficiency-Like Virus Using Capsid and Transmembrane Envelope Proteins Expressed from Recombinant Baculovirus

1999 ◽  
Vol 6 (2) ◽  
pp. 168-172 ◽  
Author(s):  
Y. Abed ◽  
G. St-Laurent ◽  
H. Zhang ◽  
R. M. Jacobs ◽  
D. Archambault
Keyword(s):  
Recombinant Protein ◽  
Fusion Protein ◽  
Western Blot ◽  
Fusion Proteins ◽  
Transmembrane Protein ◽  
Recombinant Baculovirus ◽  
Serum Samples ◽  
Western Blot Assay ◽  
Recombinant Fusion Proteins ◽  
Syncytial Virus

ABSTRACT A 120-amino-acid polypeptide selected from the transmembrane protein region (tTM) and the major capsid protein p26 of bovine immunodeficiency-like virus (BIV) were expressed as fusion proteins from recombinant baculoviruses. The antigenic reactivity of both recombinant fusion proteins was confirmed by Western blot with bovine and rabbit antisera to BIV. BIV-negative bovine sera and animal sera positive for bovine syncytial virus and bovine leukemia virus failed to recognize the recombinant fusion proteins, thereby showing the specificity of the BIV Western blot. One hundred and five bovine serum samples were tested for the presence of anti-BIV antibodies by the recombinant protein-based Western blot and a reference Western blot assay using cell culture-derived virions as test antigens. There was a 100% concordance when the p26 fusion protein was used in the Western blot. However, the Western blot using the tTM fusion protein as its test antigen identified four BIV-positive bovine sera which had tested negative in both the p26 recombinant-protein-based and the reference Western blot assays. This resulted in the lower concordance of 96.2% between the tTM-protein-based and reference Western blot assays. The results of this study showed that the recombinant p26 and tTM proteins can be used as test antigens for the serodetection of BIV-infection in animals.

scholarly journals Identification and Characterization of Avian Retroviruses in Chicken Embryo-Derived Yellow Fever Vaccines: Investigation of Transmission to Vaccine Recipients

2003 ◽  
Vol 77 (2) ◽  
pp. 1105-1111 ◽  
Author(s):  
Althaf I. Hussain ◽  
Jeffrey A. Johnson ◽  
Marcos da Silva Freire ◽  
Walid Heneine
Keyword(s):  
Western Blot ◽  
Yellow Fever ◽  
Pcr Analysis ◽  
Rt Pcr ◽  
Serum Samples ◽  
Rna Sequences ◽  
Western Blot Assay ◽  
Specific Pcr ◽  
Show Evidence

ABSTRACT All currently licensed yellow fever (YF) vaccines are propagated in chicken embryos. Recent studies of chick cell-derived measles and mumps vaccines show evidence of two types of retrovirus particles, the endogenous avian retrovirus (EAV) and the endogenous avian leukosis virus (ALV-E), which originate from the chicken embryonic fibroblast substrates. In this study, we investigated substrate-derived avian retrovirus contamination in YF vaccines currently produced by three manufacturers (YF-vax [Connaught Laboratories], Stamaril [Aventis], and YF-FIOCRUZ [FIOCRUZ-Bio-Manguinhos]). Testing for reverse transcriptase (RT) activity was not possible because of assay inhibition. However, Western blot analysis of virus pellets with anti-ALV RT antiserum detected three distinct RT proteins in all vaccines, indicating that more than one source is responsible for the RTs present in the vaccines. PCR analysis of both chicken substrate DNA and particle-associated RNA from the YF vaccines showed no evidence of the long terminal repeat sequences of exogenous ALV subgroups A to D in any of the vaccines. In contrast, both ALV-E and EAV particle-associated RNA were detected at equivalent titers in each vaccine by RT-PCR. Quantitative real-time RT-PCR revealed 61,600, 348,000, and 1,665,000 ALV-E RNA copies per dose of Stamaril, YF-FIOCRUZ, and YF-vax vaccines, respectively. ev locus-specific PCR testing of the vaccine-associated chicken substrate DNA was positive both for the nondefective ev-12 locus in two vaccines and for the defective ev-1 locus in all three vaccines. Both intact and ev-1 pol sequences were also identified in the particle-associated RNA. To investigate the risks of transmission, serum samples from 43 YF vaccine recipients were studied. None of the samples were seropositive by an ALV-E-based Western blot assay or had detectable EAV or ALV-E RNA sequences by RT-PCR. YF vaccines produced by the three manufacturers all have particles containing EAV genomes and various levels of defective or nondefective ALV-E sequences. The absence of evidence of infection with ALV-E or EAV in 43 YF vaccine recipients suggests low risks for transmission of these viruses, further supporting the safety of these vaccines.

scholarly journals Detection of Antibodies to Anaplasma phagocytophilum and Ehrlichia chaffeensis Antigens in Sera of Korean Patients by Western Immunoblotting and Indirect Immunofluorescence Assays

2003 ◽  
Vol 10 (6) ◽  
pp. 1059-1064 ◽  
Author(s):  
Jin-ho Park ◽  
Eun-jeong Heo ◽  
Kyoung-seong Choi ◽  
J. Stephen Dumler ◽  
Joon-seok Chae
Keyword(s):  
South Korea ◽  
Western Blot ◽  
Anaplasma Phagocytophilum ◽  
Fluorescent Antibody ◽  
Environmental Research ◽  
Ehrlichia Chaffeensis ◽  
Protein Antigens ◽  
Serum Samples ◽  
Western Blot Assay ◽  
Korean Patients

ABSTRACT Two hundred seventy one serum samples from South Korean patients were tested to detect antibodies against Anaplasma phagocytophilum (the human granulocytic ehrlichiosis agent) and Ehrlichia chaffeensis (the human monocytic ehrlichiosis agent) by indirect fluorescent-antibody assay (IFA) and the Western blot assay. These sera were collected from patients with symptoms of high fever. The rate of seropositivity for Orientia tsutsugamushi was 50.9% by IFA at the Public Health & Environmental Research Institute and National Institute of Health in South Korea. By IFA, 30 (11.1%) and 39 (14.4%) of the serum samples reacted with A. phagocytophilum and E. chaffeensis antigens, respectively. By the Western blot assays, 24 (8.9%) and 29 (10.7%) of the serum samples reacted with purified A. phagocytophilum and E. chaffeensis protein antigens, respectively. This report strengthens other evidence regarding the presence of A. phagocytophilum and E. chaffeensis infections in humans in South Korea.

scholarly journals Detection of Bovine Immunodeficiency Virus Antibodies in Cattle by Western Blot Assay with Recombinant Gag Protein

1997 ◽  
Vol 9 (4) ◽  
pp. 347-351 ◽  
Author(s):  
Shucheng Zhang ◽  
Wenzhi Xue ◽  
Charles Wood ◽  
Qi-Min Chen ◽  
Sanjay Kapil ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Western Blot ◽  
Bovine Immunodeficiency Virus ◽  
Test Results ◽  
Chain Reaction ◽  
Serum Samples ◽  
Western Blot Assay ◽  
Gag Protein ◽  
Immunodeficiency Virus ◽  
Polymerase Chain

A western blot assay using purified recombinant bovine immunodeficiency virus gag protein has been developed for detection of bovine immunodeficiency virus antibodies in bovine serum samples. The test was standardized with known bovine immunodeficiency virus positive and negative bovine serum samples and the monoclonal antibody to gag protein. Both naturally and experimentally infected cattle sera demonstrated positive test results. The result of western blot assay was compared with polymerase chain reaction test results in 134 blood samples collected from Kansas. Twenty-six samples tested positive for bovine immunodeficiency virus DNA with polymerase chain reaction (18.7%) and 25 were positive for the antibody to gag protein by western blot analysis (17.9%). Of 26 cattle testing positive using the polymerase chain reaction assay, 24 were antibody-positive by western blot assay, thus establishing a strong correlation between the two tests. The sensitivity and specificity of western blot relative to polymerase chain reaction are 0.92 and 0.99, respectively. The western blot assay proved to be a specific and sensitive test.

scholarly journals False-Positive Results in a Recombinant Severe Acute Respiratory Syndrome-Associated Coronavirus (SARS-CoV) Nucleocapsid-Based Western Blot Assay Were Rectified by the Use of Two Subunits (S1 and S2) of Spike for Detection of Antibody to SARS-CoV

2006 ◽  
Vol 13 (3) ◽  
pp. 409-414 ◽  
Author(s):  
Mimoun Maache ◽  
Florence Komurian-Pradel ◽  
Alain Rajoharison ◽  
Magali Perret ◽  
Jean-Luc Berland ◽  
...  
Keyword(s):  
Severe Acute Respiratory Syndrome ◽  
Western Blot ◽  
False Positive ◽  
High Sensitivity ◽  
Cross Reactivity ◽  
Serum Samples ◽  
Western Blot Assay ◽  
N Protein ◽  
False Positive Diagnosis ◽  
Healthy Donors

ABSTRACT To evaluate the reactivity of the recombinant proteins expressed in Escherichia coli strain BL21(DE3), a Western blot assay was performed by using a panel of 78 serum samples obtained, respectively, from convalescent-phase patients infected with severe acute respiratory syndrome-associated coronavirus (SARS-CoV) (30 samples) and from healthy donors (48 samples). As antigen for detection of SARS-CoV, the nucleocapsid protein (N) showed high sensitivity and strong reactivity with all samples from SARS-CoV patients and cross-reacted with all serum samples from healthy subjects, with either those obtained from China (10 samples) or those obtained from France (38 serum samples), giving then a significant rate of false positives. Specifically, our data indicated that the two subunits, S1 (residues 14 to 760) and S2 (residues 761 to 1190), resulted from the divided spike reacted with all samples from SARS-CoV patients and without any cross-reactivity with any of the healthy serum samples. Consequently, these data revealed the nonspecific nature of N protein in serodiagnosis of SARS-CoV compared with the S1 and S2, where the specificity is of 100%. Moreover, the reported results indicated that the use of one single protein as a detection antigen of SARS-CoV infection may lead to false-positive diagnosis. These may be rectified by using more than one protein for the serodiagnosis of SARS-CoV.

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