A simple and economical direct agglutination test for serodiagnosis and sero-epidemiological studies of visceral leishmaniasis

Harith, Abdallah el; Kolk, A. H. J.; Kager, Piet A.; Leeuwenburg, J; Muigai, R.; Kiugu, S; Laarman, J. J.

doi:10.1016/0035-9203(86)90149-5

Cited by 212 publications

(155 citation statements)

References 10 publications

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“…The principal phases of the procedure for preparing the DAT antigen were mass production of promastigotes of L. infantum (Iranian strain) in RPMI1640 medium supplemented with 10 % fetal bovine serum, trypsinization of the parasites, staining with Coomassie Brilliant blue and fixing with 2 % formaldehyde (Mohebali et al 2005;el Harith et al 1989;Harith et al 1986). …”

Section: Dat Testmentioning

confidence: 99%

Canine visceral leishmaniasis: seroprevalence survey of asymptomatic dogs in an endemic area of northwestern Iran

et al. 2013

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Canine visceral leishmaniasis is a major public health problem that is endemic in tropical and sub tropical countries and is fatal in humans and dogs. In addition to symptomatic dogs, asymptomatic ones seem as source of Leishmania infantum infection. Thus surveillance and control programs of reservoir hosts are essential. This study aimed to evaluate the sero-prevalence of visceral leishmaniasis in asymptomatic domestic dogs from in an endemic area of north west, Iran. A cross sectional study was carried out in Meshkin-Shahr district during 2011-2012. Blood samples collected from 508 asymptomatic domestic dogs were tested by direct agglutination test. In this study 508 dogs (397 males and 111 females, mean age, 3.24 years) from western and eastern parts of the MeshkinShahr were examined. A total of 508 dogs examined 119 dogs (23.4 %) had antibodies (titers of C1:320) against L. infantum. Statistically significance was occurred between male (25.4 %) and female (16.2) sero-prevalence (P = 0.042). No statistically significance was observed between age groups (P = 0.22). Compared with previous studies it seems to increase sero-prevalence of visceral leishmaniasis in dogs in the studied areas caused by ecological changes. High proportion of asymptomatic but seropositive dogs emphasizes the importance of dogs without clinical signs in the epidemiology of zoonotic leishmaniasis. Thus, the necessity of using serological tests in asymptomatic dogs is recommended for disease control strategy.

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Section: Dat Testmentioning

confidence: 99%

Canine visceral leishmaniasis: seroprevalence survey of asymptomatic dogs in an endemic area of northwestern Iran

et al. 2013

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“…4 El Harith and others adapted the method 5,6 according to the card-agglutination test for trypanosomiasis antigen procedures, 7 and recommended the DAT for clinical work in hospitals and health centers, 5 for seroepidemiologic studies, 5,8 and for mass VL population screening. 6,8 Several investigators reported high accuracy of the test under laboratory conditions for VL due to Leishmania donovani, [9][10][11] as well as L. infantum.…”

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confidence: 99%

Operational validation of the direct agglutination test for diagnosis of visceral leishmaniasis.

Boelaert¹,

Safi²,

Jacquet

et al. 1999

The American Journal of Tropical Medicine and Hygiene

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Abstract. The validity of the direct agglutination test (DAT) for visceral leishmaniasis (VL) was studied with a standardized field kit on 148 clinically suspected persons and 176 healthy controls recruited between 1993 and 1994 from an endemic area in Gedaref State, Sudan. A sensitivity of 95.9% and a specificity of 99.4% were found at a 1: 8,000 cut-off titer when parasitologically confirmed cases were compared with healthy controls. While corroborating previously reported sensitivity and specificity estimates of this serodiagnostic test, this study examined the bias generated by commonly used test validation procedures. The fundamental methodologic problem in VL test validation is the absence of a reliable gold standard. Moreover, any operational guideline on DAT use has to consider the critical dependency of the predictive values of the test on VL prevalence rates. The DAT diagnostic cut-off titer depends upon many external factors, among which the prevalence of disease in the area and the case mix seem the most important.

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“…The DAT was carried out essentially as described. 15 Antigen for the test was prepared from trypsin-treated, Coomassie brilliant blue-stained, formalinpreserved promastigotes of L. donovani AG83. Agglutination was carried out using a 1:500 dilution of patient sera in PBS containing 1% fetal bovine serum.…”

Section: Methodsmentioning

confidence: 99%

“…19 It is a simple technique with high sensitivity and specificity. 15,18 One of its major drawbacks is that it has no prognostic value for evaluating the progression of the disease. The DAT is used as a routine serologic test for VL in India, and the parameters of the test have been established under local conditions with very little false positivity.…”

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Serodiagnosis of leishmaniasis with recombinant ORFF antigen.

Raj¹,

Ghosh²,

Dole³

et al. 1999

The American Journal of Tropical Medicine and Hygiene

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Abstract. The serodiagnostic potential of recombinant ORFF protein (rORFF) from Leishmania infantum was assessed by ELISA. Of 49 sera from confirmed cases of visceral leishmaniasis (VL), all were seropositive using 5 ng of rORFF and serum diluted 1:20, while only 38 were positive with 500 ng of soluble antigen (SA) and 44 were positive by a direct agglutination test. There was also a positive correlation between spleen size and level of seropositivity with rORFF or SA. The reciprocal endpoint titer with rORFF was 1,280 for sera from VL patients, but Ͻ 20 with sera from malaria, filariasis, and tuberculosis patients, as well as with sera from healthy individuals from endemic and non-endemic areas. Sera from 10 confirmed cutaneous leishmaniasis cases from Turkey were negative or only weakly positive with rORFF, although 9 were positive with SA. Thus, rORFF protein appears useful as a sensitive reagent for the differential diagnosis of VL caused by the Leishmania donovani complex.Protozoan parasites of the genus Leishmania cause a spectrum of human disease ranging from self-limiting cutaneous leishmaniasis (CL) to the lethal visceral leishmaniasis (VL). There is a general correlation between the species of Leishmania and the type of disease, with the Leishmania donovani complex causing VL or kala-azar. Routine diagnosis of VL has long been based on the microscopic detection of parasites in smears of lymph node, bone marrow, or splenic aspirates or culturing the parasite from patient material. These surgical procedures for diagnosis are painful and dangerous 1 ; thus, there is an urgent need for safe and reliable tests for specific diagnosis of leishmaniasis.Serologic tests that have been developed and evaluated clinically include an immunofluorescent antibody test (IFAT), 2-4 an ELISA with either whole parasites or purified antigens, 5-10 a dot-ELISA, 11,12 immunoblot analysis, 13,14 and the direct agglutination test (DAT). [15][16][17][18] The DAT using microtiter plates for the diagnosis of VL and American CL gives satisfactory results and is easy to perform. 19 It is a simple technique with high sensitivity and specificity. 15,18 One of its major drawbacks is that it has no prognostic value for evaluating the progression of the disease. The DAT is used as a routine serologic test for VL in India, and the parameters of the test have been established under local conditions with very little false positivity. 18,20 In this paper, we report the serodiagnostic potential for Indian VL of recombinant ORFF protein (rORFF) that is encoded in the LD1 locus of chromosome 35 of L. infantum. 21 The rORFF appears to be specifically recognized by sera from patients with VL compared with those with CL. Furthermore, the sensitivity of detection appears to be greater than with total L. donovani promastigote soluble antigen, suggesting the utility of rORFF protein as a sensitive and differential diagnostic reagent. MATERIALS AND METHODSPatient sera. Sera were collected from 49 patients with clinically confirmed VL in the Sahibganj ...

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A simple and economical direct agglutination test for serodiagnosis and sero-epidemiological studies of visceral leishmaniasis

Cited by 212 publications

References 10 publications

Canine visceral leishmaniasis: seroprevalence survey of asymptomatic dogs in an endemic area of northwestern Iran

Canine visceral leishmaniasis: seroprevalence survey of asymptomatic dogs in an endemic area of northwestern Iran

Operational validation of the direct agglutination test for diagnosis of visceral leishmaniasis.

Serodiagnosis of leishmaniasis with recombinant ORFF antigen.

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