Successful substitution of fetal calf serum by human plasma for bulk cultivation of Leishmania donovani promastigotes

2013 ◽  
Vol 62 (8) ◽  
pp. 1165-1169 ◽  
Author(s):  
Abdelhafeiz Mahamoud ◽  
Hussam Ali Osman ◽  
Durria Mansour ◽  
Abdallah el Harith

The potential of human plasma (HP) or serum (HS) as a replacement for fetal calf serum (FCS) was evaluated in a liver infusion tryptose (LIT) medium for bulk cultivation of Leishmania donovani promastigotes. The promastigote yield with the LIT-FCS standard medium was 0.4–1.8×107 ml−1, and yields of 0.5–3.4×107 (P = 0.527) and 0.4–2.4×107 (P = 0.062) were recorded for two LIT medium variants containing HP or HS as supplement instead of FCS. Significantly, higher promastigote yields of 1.3–4.9×107 ml−1 were demonstrated when LIT medium was supplemented with HP of blood group O but not A, B, AB or equally pooled ABO (P = 0.007–0.020). Matching (P = 0.56) strong positive (1 : 10 2400 to ≥1 : 262 144 00) and weak negative (1 : 5–1 : 160) direct agglutination test (DAT) titres, respectively, were demonstrated in 24 visceral leishmaniasis (VL) and 45 non-VL sera for both standard LIT-FCS and alternative LIT-HP derived antigens. Our findings indicate strong potential for sustainable production of promastigotes for important diagnostic procedures such as DAT in the VL affected areas.

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 ◽  
...  

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.


2007 ◽  
Vol 14 (12) ◽  
pp. 1592-1595 ◽  
Author(s):  
Durria Mansour ◽  
Elfadil M. Abass ◽  
Mohamed el Mutasim ◽  
Abdelhafeiz Mahamoud ◽  
Abdallah el Harith

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.


2010 ◽  
Vol 18 (2) ◽  
pp. 346-348 ◽  
Author(s):  
Kamlesh Gidwani ◽  
Albert Picado ◽  
Bart Ostyn ◽  
Shri Prakash Singh ◽  
Rajiv Kumar ◽  
...  

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.


2004 ◽  
Vol 70 (1) ◽  
pp. 78-82 ◽  
Author(s):  
MOHAMMAD ZAHIDUL ISLAM ◽  
IFTIKHAR AHMED ◽  
ABDUL HALIM SARDER ◽  
RUSELLA MIRZA ◽  
EISAKU KIMURA ◽  
...  

2016 ◽  
Vol 221 ◽  
pp. 9-13 ◽  
Author(s):  
Edward Oliveira ◽  
Juliana Wilke Saliba ◽  
Diana Oliveira ◽  
Edelberto Santos Dias ◽  
Gustavo Fontes Paz

Sign in / Sign up

Export Citation Format

Share Document