The Value of a New Microculture Method for Diagnosis of Visceral Leishmaniasis by Using Bone Marrow and Peripheral Blood

Abstract

We have demonstrated that the microculture method (MCM) enables the diagnosis of visceral leishmaniasis (VL) with samples from both the bone marrow (BM) and peripheral blood (PB). The MCM is superior to the traditional culture method (TCM) as determined by its higher sensitivity in the detection of promastigotes and the more rapid time for emergence of promastigotes. The sensitivity of MCM (100% in BMs and 77.8-100% in PB) was considerably higher than that of the TCM (37.5-100% in BMs and 0-100% in PB) according to decreasing parasite density (P < 0.05). The concentration of parasites in buffy coats has increased the sensitivity of both methods, especially that of the MCM. Detection of promastigotes by MCM requires lower amounts of culture media (25-50 microL) and shorter incubation periods (2-7 days) than TCM (2.5-3.5 mL and 15-35 days, respectively). MCM was found to be valuable with the advantages of simplicity and sensitivity, in addition to being cost-effective in the routine diagnosis for VL in Adana Turkey.

Adipose Tissue-Derived Mesenchymal Stem Cells as a New Host Cell in Latent Leishmaniasis

“…In our previous studies, it has been shown that MCM is more sensitive than the other classical culture approaches in the diagnosis of visceral and cutaneous leishmaniasis. [25][26][27] It is already known that Leishmania parasites have developed a survival mechanism in which they stay viable in phagolysosome after infecting macrophages by receptor-based phagocytosis. 11 In the literature, it has been shown that parasites infect not only professional phagocytotic cells such as macrophages but also amnion epithelial cells, fibroblasts, kidney cells, and dendritic cells in mammalian hosts.…”

“…Microcapillary culture was prepared every 7 days from ADMSCs culture infected by Leishmania parasites. [25][26][27] Detected motile promastigotes were counted and evaluated according to the method of Chulay and Bryceson 24 with slight modifications: Grade 0 = 0 parasites/all fields; Grade 1+ = 1-5 parasites/all fields; Grade 2+ = 6-10 parasites/all fields; Grade 3+ = 11-25 parasites/all fields; Grade 4+ = 26-50 parasites/all fields; Grade 5+ = 51-100 parasites/all fields; Grade 6+ > 100 parasites/all fields.…”

A highly sensitive Leishmania infantum chagasi isolation method from bone marrow and peripheral blood of adults and children

“…Adaptation of this method to the microculture scale using microplates [7] can also be tested in order to facilitate the visualization of parasites during isolation period. Also, to investigate asymptomatic L. i. chagasi infection, PB can be used for culture isolation, since BM aspiration is only recommended for symptomatic patients.…”

“…Presence of macrophages in peripheral blood (PB) makes culture isolation suitable, as showed for children and adult patients infected with L. infantum from Mediterranean basin [7][8][9]. Most prominent L. infantum culture isolation techniques from PB presenting high sensitivity depends on sample washing with culture medium and concentration, or isolation of peripheral blood mononuclear cells (PBMC) [7,9], which are hard to be performed in laboratory routine, principally due to contamination risk during sample manipulation. Low efficiency of L. infantum isolation methods are probably related to immune complexes mediated parasite lysis [10,9].…”

Sensitivity of PCR and real-time PCR for the diagnosis of human visceral leishmaniasis using peripheral blood

“…Culture in specific culture medium, another technique available for parasitological diagnosis, is not routinely used, given the maintenance requirements and necessary growth periods, which can be to 30 days [27] .The lower sensitivity of culture-based methods has restricted its use, however, while fostering research into alternatives that offer improved performance [28] . Noted that bone marrow aspirate culture is not the most suitable technique for VL diagnosis, considering the time constraints it places on treatment initiation [29] .…”