Peripheral Blood Buffy Coat Smear: a Promising Tool for Diagnosis of Visceral Leishmaniasis

Abstract

Confirmative diagnosis of visceral leishmaniasis (VL) is still a challenge at the primary health care facilities in most of the rural areas of endemicity in the Indian subcontinent. Conventional methods for parasitological confirmation are risky and require skilled personnel, and hence they are unavailable to the poor people in the regions of endemicity. Buffy coat smear microscopy, as a minimally invasive, simple alternative for the parasitological diagnosis of VL, was evaluated in this prospective study. One hundred twelve VL patients were enrolled in this study. The buffy coat was separated from peripheral blood of all enrolled subjects using Histopaque-1119 solution. Leishman-stained buffy coat smears were examined for Leishmania donovani bodies, and buffy coat was also utilized for detection of parasite DNA by Leishmania nested PCR (LnPCR) for all cases. Concomitant splenic smears could be examined for L. donovani bodies in 66 cases, and the parasite load was graded on a scale of 1؉ to 6؉ for L. donovani-positive smears. All splenic smear-positive cases were also found to be positive by LnPCR. Of 112 enrolled VL cases, 103 (92%) were found to be positive for L. donovani bodies in buffy coat smear microscopy, which is promising as a confirmative diagnosis tool. We have also found a significant association of the buffy coat smear positivity with parasitic burden in the spleen smear. In this preliminary observation in Bangladesh, buffy coat smear microscopy has been found to be very simple, minimally invasive, and risk-free method of parasitological diagnosis of VL with a good diagnostic accuracy and potential for field use.

Molecular and Serological Evidence of Leishmania Infection in Stray Dogs from Visceral Leishmaniasis–Endemic Areas of Bangladesh

“…1,2 In Bangladesh, the current prevalence is estimated to be 40,000-45,000 cases with more than 40.6 million people at risk of developing the disease. 3,4 The disease is prevalent in 45 districts of Bangladesh, and most reported cases are from the Mymensingh District. 3 VL has two epidemiological patterns.…”

T cell suppression in the bone marrow of visceral leishmaniasis patients: impact of parasite load

“…The gold standard of leishmaniasis diagnosis (VL) is the microscopic demonstration of the amastigote form of parasite (LD bodies) in splenic and BM aspirates [4,20]. The visceral organs are well-known homing organs of parasites [3].…”

Efectividad de los métodos diagnósticos para la detección de ehrlichiosis monocítica humana y canina

“…Una variante de este método diagnóstico es utilizar la capa leucocitaria obtenida por centrifugación o algún otro método de concentración para la realización del frotis. Ésta ha sido utilizada para la identificación de leishmaniosis visceral, demostrando ser una opción diagnóstica para esta parasitosis; así mismo, en el caso de ehrlichiosis, los frotis realizados a partir del concentrado de leucocitos para la búsqueda de mórulas han presentado una mayor sensibilidad (66%) comparado con el frotis de sangre periférica (8%); no obstante, las mórulas se han observado con mayor frecuencia en los linfocitos que en los monocitos 24,25 .…”