TABLE 1.

NNN tube and blood agar microtiter culture technique using splenic aspirates, WBC/buffy coat cells, and PBMC

Parameter	Result for:
	NNN tube culture of splenic aspirates	Blood agar microplate culture of:
		Splenic aspirates	Buffy coat cells/WBC	PBMC
Time for detection of parasite growth (days)	5-10	3-7	5-15	3-7
No. of samples	68	68	68	47
No. of Leishmania-positive samples	67	68	58	43
Sensitivity (%)	98.5	100	85.3	91.5
Average end titer in BA plate	NAd	8.9	6.3	4.7
Dilution factor (1:3)		38.86	36.27	34.7
No. of parasites/ml specimen		∼196,830a	∼243b	∼121c

^aSA (around 100 μl) was collected in 900 μl of complete RPMI (cRPMI) medium. The 10-fold-diluted SA (150 μl) was plated in the 1st well of a 96-well microtiter plate, followed by serial 1:3 dilutions out to the 12th well.

^bWBC/buffy coat cells were separated from 3 ml of peripheral blood, resuspended in 150 μl of cRPMI medium, and plated in serial dilution (1:3) out to the 12th well.

^cPBMC were separated from 2 ml of peripheral blood, resuspended in 150 μl of cRPMI medium, and plated in serial dilution (1:3) out to the 12th well.

^dNA, not applicable.