The protozoan parasites of the genus Leishmania are the causative agents of a group of diseases
called leishmaniasis. VL is the most severe form of leishemaniasis that is almost always fatal if
untreated. Endemic leishmaniasis is endangers some 350 million people reported in a total of 98
countries most of them in the poorer regions of the globe. The burden of VL is 200,000–400,000
VL cases and 20,000–40,000 deaths per year. In Ethiopia VL have been reported from six regions
(Tigray, Amhara, Oromia, Southern Nations and Nationalities People’s Region , Somali, and Afar
regional states and The most important endemic foci include the Metema and Humera plains in
Northern Ethiopia. The objective of the present study is to assess the performance of polymerase
chain reaction (PCR) for the detection of Leishmania in microscopic negative stained tissue slide.
Non randomized purposive sampling technique was used. The genomic DNA was extracted by
using standard extraction method (the Automated Maxwell 16 instrument) and the amplifications
are using Real time PCR targeting kDNA. SPSS Version 20 was applied on the analysis of
positivity rate, Ct-values and performance of the PCR. The positive rate of Real time PCR from
microscope negative were 55 (55.6%) with a mean CT value of nearly 34+ 6.5 whereas the mean
Ct value of PCR from Known microscopic positive slides were found to be 20.22+5.38. From the
present study the performance of PCR from primarily suspect cases microscope negative slides,
test of cure and relapse cases were 34(54%) ,9(60%) and 12(57.1%) respectively, PCR has a
higher performance than Microscope, It is better to include PCR as a routine lab instrument for
those who were negative for microscope.
