Serodiagnosis of Anthroponotic Cutaneous Leishmaniasis (ACL) Caused by Leishmania tropica in Sanliurfa Province, Turkey, Where ACL Is Highly Endemic

ABSTRACT In this study, we aimed to evaluate the validity of the conventional enzyme-linked immunosorbent assay (ELISA) and the Western blotting test for the diagnosis of anthroponotic cutaneous leishmaniasis (ACL) using serum samples obtained from 51 patients with parasitologically proven nontreated CL (NonT-CL patients) and 62 patients under treatment for CL (UT-CL patients). Additionally, 29 serum samples obtained from patients with parasitologically and serologically proven visceral leishmaniasis (VL) were also used as positive controls, and serum samples from 43 blood donors were used as negative controls. All sera were diluted to the same dilution (1/100). Leishmania infantum MON-1 was used as the antigen in the conventional ELISA. The sera of 27 (93.1%) of 29 VL patients were seropositive by ELISA, while the sera of 40 (78.4%) of 51 NonT-CL patients and 43 (69.3%) of 62 UT-CL patients were seropositive by the conventional ELISA. The absorbance values of the CL patients' sera were significantly lower than the absorbance values of the VL patients' sera. Bands between 15 and 118 kDa were detected in two groups of CL patients. Among all bands, the 63-kDa band was found to be more sensitive (88.5%). When we evaluated the Western blotting results for the presence of at least one of the diagnostic antigenic bands, the sensitivity was calculated to be 99.1%. By using serological tests, a measurable antibody response was detected in most of the CL patients in Sanliurfa, Turkey. It is also noted that this response can be changed according to the sizes, types, and numbers of lesions that the patient has. The Western blot test was found to be more sensitive and valid than the conventional ELISA for the serodiagnosis of ACL. In some instances, when it is very difficult to demonstrate the presence of parasites in the smears, immunodiagnosis can be a valuable alternative for the diagnosis of ACL.

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Evidence for widespread occurrence of antibodies toEncephalitozoon cuniculi(Microspora) in man provided by ELISA and other serological tests

Parasitology ◽

10.1017/s0031182000060315 ◽

1991 ◽

Vol 102 (1) ◽

pp. 33-43 ◽

Cited By ~ 46

Author(s):

W. S. Hollister ◽

E. U. Canning ◽

A. Willcox

Keyword(s):

Western Blotting ◽

Blood Donors ◽

Pap Test ◽

Enzyme Linked Immunosorbent Assay ◽

Serological Tests ◽

Sds Page ◽

Antibody Titres ◽

Human Sera ◽

The Tropics

SUMMARYThe enzyme-linked immunosorbent assay (ELISA) was used to survey human sera for antibodies toEncephalitozoon cuniculiusing spores obtained fromin vitrocultures as antigen. Sera were obtained from patients with tropical diseases, neurological and renal disorders, patients who were HIV positive and those who had been tested for HIV but found to be negative. Sera from inhabitants of the village of Jali, The Gambia and from healthy blood donors were also examined. Numerous sera from all groups except the blood donors gave positive ELISA reactions at dilutions of 1:400. On titration, those with titres of 1:400 were reclassified as negative. Antibody titres of 1:800 and above were considered to be indicative of past or present infections withE. cuniculi. Many of these ELISA seropositives were also positive by IFAT or PAP. When examined by Western blotting of SDS–PAGE protein profiles ofE. cuniculispores, sera from many patients who had a tropical association reacted with the characteristic profiles shown by known positive mouse and rabbit sera. Others in the tropical group showed antibody binding to some but not all of the immunodominant polypeptides and yet others were negative in spite of their reactivity in the ELISA, IFAT or PAP test. Less agreement between ELISA and Western blotting results was obtained with the other groups of patients, although reactivity with one or more of the major polypeptide bands was sometimes seen. Serum from one blood donor, examined by ELISA and Western blotting, was positive. Differences in the methods of antigen preparation and of epitopes recognized by individuals may account for different reactivities in the tests. It is concluded that infections ofE. cuniculiare common in the tropics and that reactivations of these infections might be a hazard to AIDS patients.

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Serological Evaluation of Specific-Antibody Levels in Patients Treated for Chronic Chagas' Disease

Clinical and Vaccine Immunology ◽

10.1128/cvi.00106-07 ◽

2007 ◽

Vol 15 (2) ◽

pp. 297-302 ◽

Cited By ~ 38

Author(s):

Olga Sánchez Negrette ◽

Fernando J. Sánchez Valdéz ◽

Carlos D. Lacunza ◽

María Fernanda García Bustos ◽

María Celia Mora ◽

...

Keyword(s):

Chagas Disease ◽

Treatment Effectiveness ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Serological Tests ◽

Recombinant Antigens ◽

Antibody Titers ◽

Laboratory Procedures ◽

The Individual ◽

Antibody Levels

ABSTRACT Serological tests are the main laboratory procedures used for diagnosis during the indeterminate and chronic stages of Chagas' disease. A serological regression to negativity is the main criterion used to define parasitological cure in treated patients. The aim of this work was to monitor the individual specificities of antibody levels for 3 years posttreatment in 18 adult patients. Conventional serological techniques (hemagglutination assays and enzyme-linked immunosorbent assay [ELISA]) were modified by using recombinant antigens to detect early markers of treatment effectiveness. For this purpose, serum samples were taken before and during treatment and every 6 months after treatment for at least 3 years. When hemagglutination assays were used, a decrease in antibody levels was observed in only one patient. When ELISA with serum dilutions was used, antibody clearance became much more apparent: in 77.7% (14/18) of the patients, antibody titers became negative with time. This was observed at serum dilutions of 1/320 and occurred between the 6th and the 30th months posttreatment. The immune response and the interval for a serological regression to negativity were different for each patient. For some of the recombinant antigens, only 50% (9/18) of the patients reached the serological regression to negativity. Recombinant antigen 13 might be a good marker of treatment effectiveness, since 66.6% (six of nine) of the patients presented with an early regression to negativity for specific antibodies to this antigen (P = 0.002).

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Serodiagnosis of Louse-Borne Relapsing Fever with Glycerophosphodiester Phosphodiesterase (GlpQ) fromBorrelia recurrentis

Journal of Clinical Microbiology ◽

10.1128/jcm.38.10.3561-3571.2000 ◽

2000 ◽

Vol 38 (10) ◽

pp. 3561-3571 ◽

Cited By ~ 54

Author(s):

Stephen F. Porcella ◽

Sandra J. Raffel ◽

Merry E. Schrumpf ◽

Martin E. Schriefer ◽

David T. Dennis ◽

...

Keyword(s):

Serological Test ◽

Geometric Mean ◽

Eastern Africa ◽

Enzyme Linked Immunosorbent Assay ◽

Relapsing Fever ◽

Serum Samples ◽

Serological Tests ◽

Whole Cell ◽

Convalescent Phase ◽

Glycerophosphodiester Phosphodiesterase

Human louse-borne relapsing fever occurs in sporadic outbreaks in central and eastern Africa that are characterized by significant morbidity and mortality. Isolates of the causative agent,Borrelia recurrentis, were obtained from the blood of four patients during a recent epidemic of the disease in southern Sudan. TheglpQ gene, encoding glycerophosphodiester phosphodiesterase, from these isolates was sequenced and compared with the glpQ sequences obtained from other relapsing-fever spirochetes. Previously we showed that GlpQ of Borrelia hermsii is an immunogenic protein with utility as a serological test antigen for discriminating tick-borne relapsing fever from Lyme disease. In the present work, we cloned and expressed theglpQ gene from B. recurrentis and used recombinant GlpQ in serological tests. Acute- and convalescent-phase serum samples obtained from 42 patients with louse-borne relapsing fever were tested with an indirect immunofluorescence assay (IFA) and an enzyme-linked immunosorbent assay (ELISA) that used whole cells ofB. recurrentis and with immunoblotting to whole-cell lysates of the spirochete and Escherichia coli producing recombinant GlpQ. The geometric mean titers of the acute- and convalescent-phase serum samples measured by IFA were 1:83 and 1:575, respectively. The immunoblot analysis identified a high level of reactivity and seroconversion to GlpQ, and the assay was more sensitive than the whole-cell IFA and ELISA using purified, recombinant histidine-tagged GlpQ. Serum antibodies to GlpQ and other antigens persisted for 27 years in one patient. We conclude that assessment of anti-GlpQ antibodies will allow serological confirmation of louse-borne relapsing fever and determination of disease prevalence.

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Evaluation of the Serologic Cross-Reactivity between Transmissible Gastroenteritis Coronavirus and Porcine Respiratory Coronavirus Using Commercial Blocking Enzyme-Linked Immunosorbent Assay Kits

mSphere ◽

10.1128/msphere.00017-19 ◽

2019 ◽

Vol 4 (2) ◽

Cited By ~ 4

Author(s):

Ronaldo Magtoto ◽

Korakrit Poonsuk ◽

David Baum ◽

Jianqiang Zhang ◽

Qi Chen ◽

...

Keyword(s):

Culture Medium ◽

Large Deletion ◽

Cross Reactivity ◽

Enzyme Linked Immunosorbent Assay ◽

Transmissible Gastroenteritis Virus ◽

Serum Samples ◽

Serological Tests ◽

Transmissible Gastroenteritis ◽

Respiratory Coronavirus ◽

Porcine Respiratory

ABSTRACTThis study compared the performances of three commercial transmissible gastroenteritis virus/porcine respiratory coronavirus (TGEV/PRCV) blocking enzyme-linked immunosorbent assays (ELISAs) using serum samples (n = 528) collected over a 49-day observation period from pigs inoculated with TGEV strain Purdue (n = 12), TGEV strain Miller (n = 12), PRCV (n= 12), or with virus-free culture medium (n = 12). ELISA results were evaluated both with “suspect” results interpreted as positive and then as negative. All commercial kits showed excellent diagnostic specificity (99 to 100%) when testing samples from pigs inoculated with virus-free culture medium. However, analyses revealed differences between the kits in diagnostic sensitivity (percent TGEV- or PRCV-seropositive pigs), and all kits showed significant (P < 0.05) cross-reactivity between TGEV and PRCV serum antibodies, particularly during early stages of the infections. Serologic cross-reactivity between TGEV and PRCV seemed to be TGEV strain dependent, with a higher percentage of PRCV-false-positive results for pigs inoculated with TGEV Purdue than for TGEV Miller. Moreover, the overall proportion of false positives was higher when suspect results were interpreted as positive, regardless of the ELISA kit evaluated.IMPORTANCECurrent measures to prevent TGEV from entering a naive herd include quarantine and testing for TGEV-seronegative animals. However, TGEV serology is complicated due to the cross-reactivity with PRCV, which circulates subclinically in most swine herds worldwide. Conventional serological tests cannot distinguish between TGEV and PRCV antibodies; however, blocking ELISAs using antigen containing a large deletion in the amino terminus of the PRCV S protein permit differentiation of PRCV and TGEV antibodies. Several commercial TGEV/PRCV blocking ELISAs are available, but performance comparisons have not been reported in recent research. This study demonstrates that the serologic cross-reactivity between TGEV and PRCV affects the accuracy of commercial blocking ELISAs. Individual test results must be interpreted with caution, particularly in the event of suspect results. Therefore, commercial TGEV/PRCV blocking ELISAs should only be applied on a herd basis.

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

Infectious Disease Reports ◽

10.4081/idr.2020.8746 ◽

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.

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Detection of bluetongue virus antibodies in sheep from Paraná, Brazil

Semina Ciências Agrárias ◽

10.5433/1679-0359.2020v41n3p879 ◽

2020 ◽

Vol 41 (3) ◽

pp. 879

Author(s):

Maria Carolina Ricciardi Sbizera ◽

Luiz Fernando Coelho da Cunha Filho ◽

Michele Lunardi ◽

Simone Fernanda Nedel Pertile ◽

Thais Helena Constantino Patelli ◽

...

Keyword(s):

Bluetongue Virus ◽

Animal Health ◽

Enzyme Linked Immunosorbent Assay ◽

Test Results ◽

Serum Samples ◽

Agar Gel ◽

Serological Tests ◽

High Occurrence ◽

Predominant Factor ◽

World Organization

Bluetongue (BT) is an infectious and non-contagious disease caused by bluetongue virus (BTV) belonging to the genus Orbivirus. It is transmitted by a hematophagous vector, Culicoides sp., to ruminants, particularly to sheep, which are most susceptible to this disease. The main serological tests are agar gel immunodiffusion (AGID), which is recommended by the World Organization for Animal Health (OIE), and the competitive enzyme-linked immunosorbent assay (cELISA), which has the advantage of no cross-reaction with other orbiviruses. The aim was to compare the results of these two tests by conducting them on sera collected from sheep in the state of Paraná, Brazil. From March to October 2017, serum samples were collected from 270 sheep from 10 farms in six mesoregions of Paraná. The samples were subjected to AGID and cELISA to detect antibodies against BTV. Based on the test results, we classified the sheep as low, moderate, and high occurrence. The results demonstrated that 64.81% (175/270) of the sheep were seropositive through the cELISA test, showing a high occurrence, and 41.11% (111/270) were seropositive through the AGID test, indicating a moderate occurrence. The concordance between the tests was moderate (0.51) as determined by the Kappa coefficient. Among the studied farms, 90% (9/10) presented at least one seropositive sheep, and the number of animals tested positive by the cELISA test was higher than those by the AGID test. Favorable climate, which favors the presence and multiplication of the culicoid vector and the occurrence of infection, was the biggest predominant factor responsible for the obtained results. The low occurrence in farms with milder climate suggest that the presence of antibodies also occurs due to the low pathogenicity of circulating serotypes in the different mesoregions studied. It is concluded that BTV infection is present in the sheep herds in Paraná, and the occurrence was moderate detected by AGID test and high detected by cELISA test.

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Relationship of Anti-Lewis x and Anti-Lewis y Antibodies in Serum Samples from Gastric Cancer and Chronic Gastritis Patients toHelicobacter pylori-Mediated Autoimmunity

Infection and Immunity ◽

10.1128/iai.69.8.4774-4781.2001 ◽

2001 ◽

Vol 69 (8) ◽

pp. 4774-4781 ◽

Cited By ~ 25

Author(s):

Michael A. Heneghan ◽

Ciaran F. McCarthy ◽

Daiva Janulaityte ◽

Anthony P. Moran

Keyword(s):

Gastric Cancer ◽

Helicobacter Pylori ◽

Gastric Mucosa ◽

Antibody Production ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Lewis Y ◽

H Pylori ◽

Negative Controls ◽

Relationship Of

ABSTRACT Lewis (Le) antigens have been implicated in the pathogenesis of atrophic gastritis and gastric cancer in the setting ofHelicobacter pylori infection, and H. pylori-induced anti-Le antibodies have been described that cross-react with the gastric mucosa of both mice and humans. The aim of this study was to examine the presence of anti-Le antibodies in patients with H. pylori infection and gastric cancer and to examine the relationships between anti-Le antibody production, bacterial Le expression, gastric histopathology, and host Le erythrocyte phenotype. Anti-Le antibody production and H. pylori Le expression were determined by enzyme-linked immunosorbent assay, erythrocyte Le phenotype was examined by agglutination assays, and histology was scored blindly. Significant levels of anti-Lex antibody (P < 0.0001, T = 76.4, DF = 5) and anti-Ley antibody (P < 0.0001, T = 73.05, DF = 5) were found in the sera of patients with gastric cancer and other H. pylori-associated pathology compared with H. pylori-negative controls. Following incubation of patient sera with synthetic Le glycoconjugates, anti-Lex and -Ley autoantibody binding was abolished. The degree of the anti-Lex and -Leyantibody response was unrelated to the host Le phenotype but was significantly associated with the bacterial expression of Lex (r = 0.863,r 2 = 0.745, P < 0.0001) and Ley (r = 0.796,r 2 = 0.634, P < 0.0001), respectively. Collectively, these data suggest that anti-Le antibodies are present in most patients with H. pyloriinfection, including those with gastric cancer, that variability exists in the strength of the anti-Le response, and that this response is independent of the host Le phenotype but related to the bacterial Le phenotype.

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Simple and Rapid Method for Production of Whole-Virus Antigen for Serodiagnosis of Caprine Arthritis-Encephalitis Virus by Enzyme-Linked Immunosorbent Assay

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.8.2.352-356.2001 ◽

2001 ◽

Vol 8 (2) ◽

pp. 352-356 ◽

Cited By ~ 4

Author(s):

Carole Simard ◽

Molly Twinomwe Kibenge ◽

Patricia Singh ◽

Phyllis Dixon

Keyword(s):

Tissue Culture ◽

Sodium Dodecyl Sulfate ◽

Western Blotting ◽

Sodium Dodecyl ◽

Virus Antigen ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Optimum Time ◽

Caprine Arthritis Encephalitis Virus ◽

Time Of Harvest

ABSTRACT Polyethylene glycol (PEG) was used to produce whole-virus antigen derived from tissue culture cells infected with a Canadian strain of caprine arthritis-encephalitis virus. PEG antigen batches were obtained after precipitation and concentration of infected tissue culture material with PEG 8000 and final treatment with sodium dodecyl sulfate. The optimum time of harvest of tissue culture extracted material to produce the maximum amount of viral proteins was determined in roller bottles, after cocultivation of infected and noninfected fetal lamb corneal cells. Samples from day 9 to day 25 postculture were collected and processed. By Western blotting, the optimum time of harvest was found to be day 25 following the coculture. Two large batches of PEG antigen were prepared at the optimum time of harvest. Both batches gave similar results when tested by Western blotting and enzyme-linked immunosorbent assay (ELISA), using reference control sera from infected and noninfected goats. For further testing in ELISA, cutoff values and ratios were determined for PEG batch 1, using 200 known serum samples from goats free of the disease. The PEG antigen batch was compared with an in-house ELISA antigen in a kinetic mode, using 498 serum samples from field goats. The in-house ELISA antigen was produced following two rounds of ultracentrifugation and treatment with sodium dodecyl sulfate (R. A. Heckert, W. B. McNab, S. M. Richardson, and M. R. Briscoe, Can. J. Vet. Res. 56:237–241, 1992). The PEG antigen batch was found suitable for ELISA, with a relative specificity of 100% and a relative sensitivity of 99.4% compared to the in-house ELISA antigen. This method of antigen production for ELISA was found to be rapid, inexpensive, and reliable for the diagnosis of caprine-arthritis encephalitis, without requiring the use of sophisticated laboratory equipment.

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Evaluation of the performance of three serological tests for diagnosis of Leishmania infantum infection in dogs using latent class analysis

Revista Brasileira de Parasitologia Veterinária ◽

10.1590/s1984-29612020105 ◽

2020 ◽

Vol 29 (4) ◽

Author(s):

Asier Basurco ◽

Alda Natale ◽

Katia Capello ◽

Antonio Fernández ◽

María Teresa Verde ◽

...

Keyword(s):

Sensitivity And Specificity ◽

Leishmania Infantum ◽

Latent Class ◽

Rapid Test ◽

Antibody Test ◽

Latent Class Models ◽

Canine Leishmaniasis ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Serological Tests

Abstract Canine leishmaniasis (CanL) is a disease caused by Leishmania infantum. Serological methods are the most common diagnostic techniques used for the diagnosis of the CanL. The objective of our study was to estimate the sensitivity and specificity of one in-house ELISA kit (ELISA UNIZAR) and three commercially available serological tests (MEGACOR Diagnostik GmbH) including an immunochromatographic rapid test (FASTest LEISH®), an immunofluorescent antibody test (MegaFLUO LEISH®) and an enzyme-linked immunosorbent assay (MegaELISA LEISH®), using latent class models in a Bayesian analysis. Two hundred fifteen serum samples were included. The highest sensitivity was achieved for FASTest LEISH® (99.38%), ELISA UNIZAR (99.37%), MegaFLUO LEISH® (99.36%) followed by MegaELISA LEISH® (98.49%). The best specificity was obtained by FASTest LEISH® (98.43%), followed by ELISA UNIZAR (97.50%), whilst MegaFLUO LEISH® and MegaELISA LEISH® obtained the lower specificity (91.94% and 91.93%, respectively). The results of present study indicate that the immunochromatographic rapid test evaluated FASTest LEISH® show similar levels of sensitivity and specificity to the quantitative commercial tests. Among quantitative serological tests, sensitivity and specificity were similar considering ELISA or IFAT techniques.

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Detection of Specific Antibodies to an Antigenic Mannoprotein for Diagnosis of Penicillium marneffeiPenicilliosis

Journal of Clinical Microbiology ◽

10.1128/jcm.36.10.3028-3031.1998 ◽

1998 ◽

Vol 36 (10) ◽

pp. 3028-3031 ◽

Cited By ~ 42

Author(s):

Liang Cao ◽

Da-Liang Chen ◽

Cindy Lee ◽

Che-Man Chan ◽

King-Man Chan ◽

...

Keyword(s):

Specific Antibody ◽

Blood Donors ◽

Antibody Test ◽

High Specificity ◽

Pathogenic Fungi ◽

Enzyme Linked Immunosorbent Assay ◽

Serum Samples ◽

Specific Antibodies ◽

Immunodeficiency Virus ◽

Positive Results

The disseminated and progressive fungal disease Penicillium marneffei penicilliosis is one of the most common infectious diseases in AIDS patients in Southeast Asia. To diagnose systemic penicilliosis, we developed an enzyme-linked immunosorbent assay (ELISA)-based antibody test with Mp1p, a purified recombinant antigenic mannoprotein of P. marneffei. Evaluation of the test with guinea pig sera against P. marneffei and other pathogenic fungi indicated that this assay was specific for P. marneffei. Clinical evaluation revealed that high levels of specific antibody were detected in two immunocompetent penicilliosis patients. Furthermore, approximately 80% (14 of 17) of the documented penicilliosis patients with human immunodeficiency virus tested positive for the specific antibody. No false-positive results were found for serum samples from 90 healthy blood donors, 20 patients with typhoid fever, and 55 patients with tuberculosis, indicating a high specificity of the test. Thus, this ELISA-based test for the detection of anti-Mp1p antibody can be of significant value as a diagnostic for penicilliosis.

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