scholarly journals The rK39 Antigen from an Iranian Strain of Leishmania infantum: Detection of Anti-Leishmania Antibodies in Humans and Dogs

2020 ◽  
Author(s):  
Bibi Razieh HOSSEINI FARASH ◽  
Mehdi MOHEBALI ◽  
Bahram KAZEMI ◽  
Abdolmajid FATA ◽  
Reza RAOOFIAN ◽  
...  
Keyword(s):  
Leishmania Infantum ◽  
Field Studies ◽  
Severe Form ◽  
Recombinant Antigen ◽  
Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Controls ◽  
Direct Agglutination Test ◽  
Human Sera ◽  
Extended Field

Background: Visceral leishmaniasis (VL) is the most severe form of leishmaniasis in Iran with high mortality rates in the case of inaccurate diagnosis and treatment. This study aimed to prepare and evaluate a new rk39 recombinant antigen from an Iranian strain of Leishmania infantum for diagnosis of VL in humans and dogs. Methods: rK39-based enzyme-linked immunosorbent assay (ELISA) was compared with the direct agglutination test (DAT) for the detection of anti L. infantum antibodies. We screened 84 human sera and 87 dog sera from clinical cases in the endemic area of Meshkin-Shahr, Iran along with 176 sera from healthy controls (collected from 86 humans and 90 dogs) during 2013 -2016. Results: Using the rK39 ELISA, a sensitivity of 85.7% (95% CI, 95-99%) and a specificity of 86.0% (95% CI, 95%-99%) were detected in human sera at a 1:800 (cut-off) titer when DAT-confirmed cases were compared with healthy controls; a sensitivity of 96.6% (95% CI, 95%-99%) and specificity of 94.4% (95% CI, 95%-99%) were found at a 1:80 (cut-off) titer compared with DAT. Kappa analysis indicated agreement between the rK39 ELISA and DAT (0.718) when using human sera at a 1:800 (cut-off) titer as well as (0.910) at a 1:80 (cut-off) titer when using dog sera (P<0.05). Conclusion: New rk39 recombinant antigen from an Iranian strain of Leishmania infantum seems to be used for diagnosis of VL in humans and dogs. Further extended field studies are recommended

Development and evaluation ofLeishmania infantumrK26 ELISA for serodiagnosis of visceral leishmaniasis in Iran

Parasitology ◽  
2008 ◽  
Vol 135 (9) ◽  
pp. 1035-1041 ◽  
Author(s):  
S. FARAJNIA ◽  
B. DARBANI ◽  
H. BABAEI ◽  
M. H. ALIMOHAMMADIAN ◽  
F. MAHBOUDI ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Infectious Diseases ◽  
Leishmania Infantum ◽  
Elisa Test ◽  
Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Agglutination Test ◽  
Whole Cell ◽  
Reliable Diagnosis ◽  
Cell Elisa

SUMMARYThe purpose of this study was to prepare recombinant K26 antigen fromLeishmania infantumand evaluate its performance by enzyme-linked immunosorbent assay (ELISA) test for serodiagnosis of visceral leishmaniasis (VL) in endemic regions of Iran. The results were compared with those obtained by direct agglutination test (DAT) and whole cell ELISA using crude parasite antigen. Of 93 sera from patients with confirmed VL, 90 sera were positive with rK26 ELISA (sensitivity=96·8%), whereas 85 sera were positive with DAT (sensitivity=91·4%) and 89 sera were positive with whole cell ELISA (sensitivity=95·7%). Of 130 subjects who either had other infectious diseases (n=30) or were healthy (n=100), rK26 ELISA were negative in all cases (specificity=100%), whereas DAT were negative in 116 cases (specificity=89·2%) and whole cell ELISA was negative in 114 cases (specificity=87·7%). The results of this study indicate that the rK26 ELISA is more sensitive and specific than conventional methods and could be used for reliable diagnosis of VL caused byLeishmania infantum.

scholarly journals Diagnosis of Leishmania infantum using Direct Agglutination Test and rKE16 Dipstick Rapid Test in Domestic Dogs from Ardabil Province, Iran

2015 ◽  
Vol 10 (3) ◽  
pp. 102-110
Author(s):  
H. Nahrevania ◽  
P. Ghaffarine ◽  
M. Farahmand ◽  
M. Mohebali ◽  
F. Zaboli ◽  
...  
Keyword(s):  
Leishmania Infantum ◽  
Rapid Test ◽  
Agglutination Test ◽  
Domestic Dogs ◽  
Direct Agglutination Test

scholarly journals Comparison of Enzyme-Linked Immunosorbent Assay, Indirect Fluorescent Antibody Test, and Direct Agglutination Test for Detecting Toxoplasma Gondii Antibodies in Naturally Aborted Ovine Fetuses

1989 ◽  
Vol 1 (2) ◽  
pp. 124-127 ◽  
Author(s):  
Sheryl L. Seefeldt ◽  
Clyde A. Kirkbride ◽  
Jitender P. Dubey
Keyword(s):  
Toxoplasma Gondii ◽  
Immunosorbent Assay ◽  
Indirect Fluorescent Antibody Test ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Agglutination Test ◽  
Indirect Fluorescent Antibody ◽  
Special Equipment

Results obtained in an enzyme-linked immunosorbent assay (ELISA), an indirect fluorescent antibody test (IFA), and a modified direct agglutination test (MAT) for Toxoplasma gondii antibodies from examination of fetal fluids from 377 aborted ovine fetuses were compared. Sixty-seven samples were positive by MAT (titers 1:16 to > 1:65,536), 58 were positive by ELISA, and 62 were positive by immunoglobulin G-IFA. The MAT was preferred because it required less time, labor, and special equipment. It was simple to run, could be done on serum from any species without modification, and it was more effective than the IFA for detecting toxoplasma antibodies in severely autolyzed fetuses. No advantage was found in determining immunoglobulin M antibodies in ovine fetal sera.

scholarly journals Performance of an indigenous β-mercaptoethanol-modified antigen in comparison with a commercial reference in direct agglutination test for detection of canine visceral leishmaniasis

2014 ◽  
Vol 63 (1) ◽  
pp. 106-110
Author(s):  
Saul José Semião-Santos ◽  
Laura Barral Veloso ◽  
Paulo Paes de Andrade ◽  
Marcia Almeida de Melo ◽  
Luis Miguel Lourenço Martins ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Leishmania Infantum ◽  
Leishmania Donovani ◽  
Agglutination Test ◽  
Canine Visceral Leishmaniasis ◽  
Direct Agglutination Test ◽  
Serum Samples

We compared the performance of a locally produced β-mercaptoethanol-modified promastigote antigen (β-ME-Ag) of an indigenous Leishmania infantum strain against that of a trypsinized Leishmania donovani reference (REF-Ag) in the direct agglutination test (DAT) for detection of canine visceral leishmaniasis (CVL). One hundred and fifty-one serum samples collected from dogs belonging to four groups with different conditions were included. At a DAT titre of 1 : 320, statistically determined as optimal cut-off value for β-ME-Ag, and 1 : 160 for REF-Ag, a sensitivity and a specificity of 100 % were estimated for β-ME-Ag in comparison with 96.6 % and 100 %, respectively, for REF-Ag. Overall, levels of agglutination titres recorded for the two antigens were highly concordant (Cohen’s κ = 0.879) in both the CVL and non-CVL groups. Based on current results, and ease experienced in processing the antigen and reading the test outcome, we recommend incorporation of β-ME-Ag in DAT for confirmation or exclusion of suspected CVL in dogs.

scholarly journals Use of a Newly Developed β-Mercaptoethanol Enzyme-Linked Immunosorbent Assay To Diagnose Visceral Leishmaniasis in Patients in Eastern Sudan

2007 ◽  
Vol 14 (12) ◽  
pp. 1592-1595 ◽  
Author(s):  
Durria Mansour ◽  
Elfadil M. Abass ◽  
Mohamed el Mutasim ◽  
Abdelhafeiz Mahamoud ◽  
Abdallah el Harith
Keyword(s):  
Visceral Leishmaniasis ◽  
Immunosorbent Assay ◽  
Leishmania Donovani ◽  
Clinical Evidence ◽  
High Reliability ◽  
Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Agglutination Test ◽  
Symptomatic Group ◽  
Eastern Sudan

ABSTRACT Corroboration of serology results is essential for restricting the risk of inappropriate antileishmanial prescription. A direct agglutination test (DAT) and a recently developed β-mercaptoethanol-modified enzyme-linked immunosorbent assay (β-ME ELISA) based on the use of antigen prepared as described for the DAT were applied to 416 sera from two Sudanese populations with and without clinical evidence of visceral leishmaniasis (VL). Of 285 sera with the lowest antileishmanial DAT titers (≤1:100 to 1:1,600), 270 (94.7%) scored comparable minimum β-ME ELISA absorbance values (≤0.1 to 0.26). In 117 sera that demonstrated the highest DAT titers (1:12,800 to ≥1:25,600), 86 (73.5%) scored maximum (0.81 to ≥1.35) and 30 (25.6%) medium (0.27 to 0.80) β-ME ELISA absorbance values. VL diagnosis was established for 142 (44.1%) patients in the VL-symptomatic group (n = 322), based on positive microscopy for Leishmania donovani in lymph node aspirates or positive DAT (titer, ≥1:3,200). Of the 125 sera from the symptomatic patients for whom microscopy was positive for VL, 111 (88.8%) had comparable positive DAT and β-ME ELISA readings. In all 17 sera from the symptomatic DAT-positive patients for whom leishmaniasis was not established by microscopy but who responded favorably to antileishmanial therapy, absorbance values (≥0.27) indicative of VL were obtained by β-ME ELISA. Of 197 symptomatic patients for whom microscopy was negative for VL, 172 (87.3%) tested negative in β-ME ELISA and 180 (91.4%) in DAT. Based on the high reliability demonstrated here for VL detection, β-ME ELISA fulfills the requirement of confirming DAT results in patients manifesting suspected VL.

scholarly journals Seroprevalence of Toxoplasma gondii and Leishmania spp. in domestic donkeys from Portugal

2019 ◽  
Vol 28 (1) ◽  
pp. 172-176 ◽  
Author(s):  
Filipa Teixeira Rodrigues ◽  
Catarina Pereira ◽  
Jitender Prakash Dubey ◽  
Miguel Nóvoa ◽  
Miguel Quaresma ◽  
...  
Keyword(s):  
Toxoplasma Gondii ◽  
Leishmania Infantum ◽  
Agglutination Test ◽  
Direct Agglutination Test ◽  
Serum Samples ◽  
Modified Agglutination Test ◽  
Leishmania Spp

Abstract Toxoplasma gondii and Leishmania infantum are zoonotic protozoal parasites. Serum samples were obtained from 186 donkeys (Equus africanus asinus) from Portugal and assessed for antibodies to T. gondii by the modified agglutination test (MAT). For titration of antibodies to Leishmania spp. the direct agglutination test was used (DAT). Eleven donkeys were seropositive for T. gondii with titres of 20 (n = 7), 80 (n = 2), 640 (n = 1) and ≥ 2560 (n = 1). One donkey was seropositive for Leishmania spp. (titre of 800). Donkeys in Portugal are exposed to and can be infected with T. gondii and Leishmania spp.

scholarly journals Seroprevalence of brucellosis in patients with prolonged fever in Bangladesh

2016 ◽  
Vol 10 (09) ◽  
pp. 939-946 ◽  
Author(s):  
AKM Anisur Rahman ◽  
Dirk Berkvens ◽  
Claude Saegerman ◽  
David Fretin ◽  
Noor Muhammad ◽  
...  
Keyword(s):  
Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Brucellosis ◽  
Chain Reaction ◽  
Serological Tests ◽  
Human Sera ◽  
Polymerase Chain ◽  
Species Specific ◽  
First Time ◽  
Sera Samples

Introduction: This study describes the seroprevalence of human brucellosis among pyretic patients and detection of Brucella abortus DNA from seropositive pyretic patients using real-time polymerase chain reaction (rtPCR) for the first time in Bangladesh. Methodology: Blood samples were collected from 300 pyretic patients from October 2007 to May 2008 and subjected to three serological tests: Rose-Bengal plate test (RBT), standard tube agglutination test (STAT), and indirect enzyme-linked immunosorbent assay (iELISA). Risk factors were identified by multivariate Firth’s logistic regression analysis. Brucella genus (BCSP31) and species-specific (IS711) rtPCR were applied to six human sera samples. Results: The seroprevalence of brucellosis among pyretic patients was estimated to be 2.0% (95% confidence interval [CI]: 0.74–4.30). The odds of brucellosis seropositivity were 8.9 (95% CI: 1.26–63.0) times higher in pyretic patients who handled goats than those who handled only cattle, whereas the odds of brucellosis seropositivity were 9.7 (95% CI: 1.28–73.68) times higher in pyretic patients who had backache compared to those without backache. B. abortus DNA was amplified from all six human sera that tested positive by RBT, STAT, and iELISA. As the agreement between the tests was very strong, RBT is recommended as a screening test for the diagnosis of human brucellosis in Bangladesh because it is easier to use, cheaper, and faster. Conclusions: Brucellosis among pyretic patients is common, and B. abortus is responsible for brucellosis in such patients. Pyretic patients who handle goats and those with backaches should be screened for brucellosis.

scholarly journals Persistence ofLeishmania donovaniAntibodies in Past Visceral Leishmaniasis Cases in India

2010 ◽  
Vol 18 (2) ◽  
pp. 346-348 ◽  
Author(s):  
Kamlesh Gidwani ◽  
Albert Picado ◽  
Bart Ostyn ◽  
Shri Prakash Singh ◽  
Rajiv Kumar ◽  
...  
Keyword(s):  
Visceral Leishmaniasis ◽  
Immunosorbent Assay ◽  
Leishmania Donovani ◽  
Extended Period ◽  
Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Agglutination Test ◽  
History Of ◽  
Antibody Titers

ABSTRACTThe persistence of anti-Leishmania donovaniantibodies in past visceral leishmaniasis (VL) cases was retrospectively assessed by means of the direct agglutination test (DAT) and the rK39 enzyme-linked immunosorbent assay (ELISA). Antibody titers remained high for an extended period of time in past cases of VL. These results highlight the need to carefully elicit the history of patients with VL symptoms.

scholarly journals A Modified Agglutination Test for Neospora caninum: Development, Optimization, and Comparison to the Indirect Fluorescent-Antibody Test and Enzyme-Linked Immunosorbent Assay

1998 ◽  
Vol 5 (4) ◽  
pp. 467-473 ◽  
Author(s):  
Andrea E. Packham ◽  
Karen W. Sverlow ◽  
Patricia A. Conrad ◽  
Emily F. Loomis ◽  
Joan D. Rowe ◽  
...  
Keyword(s):  
Indirect Fluorescent Antibody Test ◽  
Neospora Caninum ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Agglutination Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Direct Agglutination Test ◽  
Indirect Fluorescent Antibody ◽  
Secondary Antibodies ◽  
Higher Sensitivity

ABSTRACT Current serologic tests used to detect antibodies to Neospora caninum require species-specific secondary antibodies, limiting the number of species that can be tested. In order to examine a wide variety of animal species that may be infected with N. caninum, a modified direct agglutination test (N-MAT) similar to the Toxoplasma gondii modified direct agglutination test (T-MAT) was developed. This test measures the direct agglutination of parasites by N. caninum-specific antibodies in serum, thus eliminating the need for secondary host-specific anti-isotype sera. The N-MAT was compared to the indirect fluorescent-antibody test (IFAT) and the enzyme-linked immunosorbent assay (ELISA) with a “gold standard” serum panel from species for which secondary antibodies were available (n = 547). All positive samples tested were from animals with histologically confirmed infections. Up to 16 different species were tested. The N-MAT gave a higher sensitivity (100%) and specificity (97%) than the ELISA (74 and 94%, respectively) and had a higher sensitivity but a lower specificity than the IFAT (98 and 99%, respectively). The reduced specificity of the N-MAT was due to false-positive reactions in testing fetal fluids with particulate matter or severely hemolyzed serum. Overall, the N-MAT proved to be highly sensitive and specific for both naturally and experimentally infected animals, highly reproducible between and within readers, easy to use on large sample sizes without requiring special equipment, and useful in testing serum from any species without modification.

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