Peripheral blood buffy coat smear : A promising tool for diagnosis of visceral

11 Confirmative diagnosis of Visceral Leishmaniasis (VL) is still being a challenge at the primary 12 health care facilities in most of the rural endemic areas in the Indian sub-continent. Conventional 13 methods for parasitological confirmation are risky and require skilled personnel hence, 14 unreachable to the poor people in the endemic region. Buffy coat smear microscopy, as a 15 minimally invasive simple alternative, for the parasitological diagnosis of VL was evaluated in 16 this prospective study. One hundred twelve VL patients were enrolled in this study. Buffy coat 17 was separated from peripheral blood of all enrolled subjects using the Histopaque-1119 solution. 18 Leishman-stained buffy coat smear was examined for Leishmania Donovani (LD) body and 19 buffy coat was also utilized for detection of parasite DNA by Leishmania-nested polymerase 20 chain reaction (Ln-PCR) for all cases. Concomitant splenic smears could be examined for LD 21 body in 66 cases and parasite load was graded in a scale of 1+ to 6+ for LD-positive smears. All 22 Copyright © 2011, American Society for Microbiology. All Rights Reserved. J. Clin. Microbiol. doi:10.1128/JCM.05067-11 JCM Accepts, published online ahead of print on 28 December 2011 2 11 January 2012 on O cber 2, 2017 by gest ht://jcm .sm .rg/ D ow nladed fom splenic smear-positive cases were also found positive by Ln-PCR. Of 112 enrolled VL cases, 103 23 were found positive (92%) for LD bodies in buffy coat smear microscopy which is promising as 24 a confirmative diagnosis tool. We have also found a significant association in the buffy coat 25 smear positivity with parasitic burden in the spleen smear. In this preliminary observation in 26 Bangladesh, buffy coat smear microscopy has been found to be very simple, minimally invasive 27 and risk-free method of parasitological diagnosis for VL with a good diagnostic accuracy for its 28 potentiality in field use. 29